CN1871058A - Multiple laminar flow-based particle and cellular separation with laser steering - Google Patents

Abstract

Here is a device (100) for separating fluid multi components (A, B, C, D). The device is comprised of an optically transparent screen separation channel (110), the first import (120) for the first fluid flow (W), the second import (120) for the second fluid flow (X), the first export (130) for the first fluid flow (W), the second export (130) for the second fluid flow (X), holographic optical trap system (210) joint to optically transparent screen separation channel (210). Wherein the first component A of the multi components (A, B, C, D) in the first fluid flow (W) can be selectively transferred to the second fluid flow (X) and engender enriched the second fluid flow (X).

Description

Utilize laser steering to carry out particle and the cell separation that flows based on multilayer

Technical field

The present invention relates generally to and is used for separating particles or material; for example with blood; sperm and other particle go out their different compositions and the technology and the system that separate with cell separation; this technology and system use multilayer stream and further combine with laser steering, for example are used in combination with holographic optics trapping and manipulation.

Background technology

Haemocyte has several classification.The amount of red blood cell or red blood cell (RBC) is 4.8 hundred ten thousand cells/μ l to the woman, is 5.4 hundred ten thousand cells/μ l to the man.RBC constitutes the solid constituent of blood 93% and accounts for about 42% of volumetric blood.Blood platelet is 2 μ m-3 μ m sizes, and their constitute the solid constituent of blood 7% and account for the about 3% of volumetric blood, are equivalent to every liter about 1.5 to 4 * 10 ¹¹Individual cell.Leucocyte (WBCs) or leucocyte that 5 kinds of general types are arranged, every liter about 1.5 to 4 * 10 of quantity ⁹Individual cell.WBCs comprises 50-70% neutrophil cell (12 μ m-15 μ m size); 2-4% eosinocyte (12 μ m-15 μ m size); 0.5-1% basocyte (9 μ m-10 μ m size); 20-40% lymphocyte (25% B cell and 75% T cell) (8 μ m-10 μ m size); And 3-8% monocyte (16 μ m-20 μ m size).They comprise the solid constituent of blood 0.16% and account for volumetric blood near 0.1%, be equivalent to every liter about 4 to 12 * 10 ⁹Individual cell.The amount of the object WBC that infects can be up to every liter 25 * 10 ⁹

Blood platelet is a cell minimum in the blood, and its protein that discharges in blood when solidifying is important.The patient who causes this quantity to reduce the immunological diseases of (for example tumour, leukaemia and other patients undergoing chemotherapy) needs blood platelet to inject to become low with the quantity that prevents them sometimes.Adult's orthoplastocyte quantity is 140, and 000-440 between 000 cell/μ l, is not lower than 50,000 cells/μ l and should numeral should not drop to, because blood platelet is played a leading role in blood clotting.

The blood isolation technics has adopted discontinuous centrifugal process to handle traditionally.More specifically, take out the blood of a constant volume from donor at special time.The blood of this volume is accepted the centrifugal force of varying level, separates so that corresponding blood to blood constituent to be provided, for example blood plasma, blood platelet, red blood cell and leucocyte.Such processing is discontinuous rather than continuous, like this if get more blood processing from donor, will take out the blood of another volume from donor, and process repeats.

The step that blood platelet is collected is: collect blood from donor; Add anticoagulant; Separate by centrifugal force; Red blood cell, white blood cell and blood plasma are returned donor.Once collection comprises 200-400ml blood plasma, reduces it to avoid incompatibility.This collection comprises about 8.5 * 10 usually ¹⁰Blood platelet.Although more blood platelet can be separated from blood, donor is dedicated his/her usually to hematoblasticly can not lose its blood coagulation ability near 10%.These blood platelets must be used up in five days that collect.

Blood platelet exclusion (being called external removing method (apheresis)) is existing processing method [Haemonetics Component Collection System (CCS) and Multi Component System (the Multi) (Haemonetics that blood platelet is separated, Braintree, MA)].This automatic machinery separates blood platelet from blood will continue 1.5 to 2 hours time (amount of donating blood of supposition 10%).This handles the fast and full automation than traditional method, and can be used as one or twice blood platelet dosage.Yet this processing still is slow with respect to patient's patience, and is possible to the improvement that separates the purity that blood platelet separates.

Other processing method also is consuming time, often will spend several hrs, particularly separates will fail back donor the time when blood useless.For example, blood platelet is contributed to make and will be spent several hrs, and whole blood is extracted out from donor, obtains blood platelet by centrifugation, and remaining blood constituent is then injected back donor.This centrifugal treating also is more coarse, also can cause and gather a certain proportion of destruction of cell, obviously reduces the output that useful blood separates.

Other type of separation also is consuming time or can not handles wide variety of materials within a certain period of time.For example, the screening of sperm (sperm that wherein will move about or activated is isolated with sperm that do not move about or non-activity) is work very consuming time, and it is subjected to strict capacity limit.

The present invention is such as described in greater detail, to the manipulation (for example in second and the 5th related application, describing) of particle, and the also part of novel isolation technics.A conventional art is the optics trapping in handling micro-object.The acceptable description of optics trapping effect is that the light (for example light that is focused on by high-NA microlens group) of tight focus has intensity gradient jumpy.Optical trap uses the gradient force of light beam according to its dielectric constant trapping particle.

In order to minimize its energy, the high particle of permittivity ratio surrounding medium will move on to the optical trap zone, and electric field is the strongest herein.Have at least that with dielectric constant around it particle of fine difference to be arranged be responsive to this gradient, and relatively highlight strength point is attracted or is ostracised, promptly near or away from beam focus.In the optical trap of structure, the optical gradient forces that is produced by single beam is handled the position of dielectric grain, and this particle immerses in the fluid media (medium) that has less than the particle refractive index, but also handles the particle of refraction, absorption and low-k.

Optical gradient forces in the optical trap and the radiation pressure competition that trends towards moving the particle of trapping along beam axis.Optical trap can be placed on object lens focusing space Anywhere by the direction of propagation and the collimation of selecting input beam approx.Collimated light beam is injected the focus at the back aperture arrival lens focussing plane center of object lens, and eccentric focus is injected and arrived to another light beam with an angle simultaneously.The light beam of dispersing a little focuses on the focussing plane downstream and the light beam assembled focuses on the upstream.Multi-beam is injected the pupil of lens simultaneously, and each is focusing on optical trap of space formation, and its position is by its incident angle decision.Holographic optics trapping technology uses phase place modulation diffraction optical element to force the phase diagram with multi-beam to be added on the wave surface of single input light, with this single beam is become multi-beam.

The modulation of the phase place of incident beam is preferably used for producing optical trap, because trapping depends on beam intensity and do not rely on their relative phase.Modulation and Amplitude Modulation can shift light and reduce its effect from trap.

When optics trapping particle, the optical gradient forces that is applied by trap surpasses from other radiation pressure of scattering and absorption rising.For Gauss TEM _OoInput laser beam, this means that usually beam diameter should conform to the inlet pupil basically.The preferred minimum value aperture about 0.9 to about 1.0 that forms trap.

In a difficult point implementing optics trapping technology is that each trap that produces usually requires it to have the light beam of focusing.The a plurality of optical traps of interested many system requirements have improved several and have been used to realize the method for a plurality of structure of traps.An existing method uses single light beam to change direction between a plurality of trap location, with " time shares " this Shu Guang between each trap.Yet along with the increase of trap quantity, elongated in the interval of each trap " pass " state, particle just spread out from trap before trap powers up again thus.All these relations are limited in each system less than 10 traps with the enforcement of this method.

Another conventional method that produces a plurality of trap system depends on a plurality of light beams and passes single high numerical aperture lens group simultaneously.This is by using multiple laser or using one or more optical splitters to finish in single laser beam.A problem of this technology is that along with the increase of trap quantity, optical system becomes and little by little becomes increasingly complex.Because these problems, the enforcement of known this method are limited in each system less than about 5 traps.

In the 3rd method that realizes many trap system, a diffraction optical element (DOE) (for example utilizing the phase shift hologram of one of transmission or reflection geometry) is used to change the wave surface of single laser beam.This disclosure of the Invention is in people's such as Grier U.S. Patent No. 6,055,106.Change wave surface, make the laser beam in downstream become a large amount of single laser beams basically, and the relative position and the direction of propagation of being determined by the essence of diffraction optical element arranged.In fact, the Fourier transform of DOE produces one group of light intensity crest, and each is as independent trap or " light pincers ".

Some of the third method implements to have used a fixing transmission hologram to create independent trapping center between 16 to 400.

Used fixed hologram figure to confirm the principle of holographic optics trapping, but used liquid crystal grating as the hologram that is used for the independent hologram " production " of the new distribution of each trap.The modulation of the phase place of spatial variations is applied on the trapping laser by liquid crystal grating, and this can be easily by real time computer control, thereby allows various dynamic manipulations.

Can use the trap of other type to come optics trapping particle, include but not limited to optical vortex, optical bottle, optical rotator and optics cage.Optical vortex produces the gradient around zero electric field region, and it has dielectric constant to manipulation, and to be lower than other type of particle particle of surrounding medium or reflection or that repelled by optical trap useful.In order to minimize its energy, this particle will move on to the minimum zone of electric field, be exactly at the zero electric field region at the focus place of moulding laser beam suitably.Optical vortex provides zero electric field region the spitting image of the hole in annulus (annular).Optical gradient is radially, and its maximum electric field is at the periphery of annulus.Optical vortex is trapped in granule in the hole of annulus.Can finish delay by making vortex slide past granule along zero electric field line.

Optical bottle is different from the optical vortex part and is, it just has zero electric field and have non-zero electric field at vortex one end ring around all other directions of focus in focus.Optical bottle is useful aspect trapping atom and micelle, because these particles are too little or too absorbent can not with the trapping of optical vortex or light pincers (see J.Arlt and M.J.Padgett. " Generation of a beam with adark focus surrounded by regions of higher intensity:The optical bottlebeam; " Opt.Lett.25,191-193,2000).

Optics cage (U.S. Patent No. 5,939,716) is the similar of the macroscopic optical vortex of loosely.The optics cage forms the time average ring of optical trap, with around too big or produce reflection be lower than the particle that surrounding medium can not trapping with dielectric constant.

When laser beam by direct projection of phase diagram optical element or reflex time, the phase diagram optical element produces has a plurality of penlights with variable phase profile.According to the quantity and the type of required optical trap, change can comprise diffraction, wave surface finishing, phase transition, turn to, disperses and assemble.Select based on the phase place profile, can use the phase diagram optical element to produce optical trap, this optical trap can be with two or more combination in optical trap, optical vortex, optical bottle, optical rotator, optics cage and these forms.

The researcher has sought to be used for the indirect method of manipulated cell, and for example mark has the cell of rhombus particulate and then clamps and obtain the rhombus particulate.Cell operation comprises celluar localization and the tensile cell that is used for micro-analysis.Histocyte is also used up pincers outward by organism and is arranged, arranges as same in vivo space.

Except that cell itself, the light pincers have been used for manipulated cell device (as bubble, chromosome or the spherical DNA that transmits along microtubule).Make and use up pincers and also object can be inserted cell.

Therefore,, need a kind of uses from other cell as the example of the optical trap screening newtype that utilizes laser steering, organize with pollutant in separate the method that the technology of valuable cell is carried out cell, screening.Further, need a kind of uniqueness contribution that realize the optics trapping with the industrial requirement that satisfies a large amount of haemocytes screenings and the method for purification needs.Also have, at animal feeding market demand separated sperm cell.

As a result, still need continuous, that have high-throughput, time-saving and the various compositions that will separate are caused the technology and the device of the separation of negligible or microlesion.In addition, this technology should have further using value to biology or medical domain, for example is used for separating blood, sperm, other cell material and virus, organelle, chondritic, colloid suspended substance and other biomaterial.

Summary of the invention

Exemplary embodiment of the present provides and has been used at the mixture separated component, for example the different blood constituents of whole blood is separated into accordingly to separate, and for example blood platelet separates, red blood cell separates, leucocyte separates and blood plasma separates.Various embodiments of the invention provide the separation of carrying out composition in the continuous substrate (basis) (for example within the continuous closed system), and the potential damage and the pollution that art methods are not caused, particularly for the blood constituent partition method.Continuous processing procedure of the present invention has also effectively been saved the time of blood separation and higher output is provided.In addition, different embodiment also comprises and is used to separate and handles composition, particularly holographic optics and handle and the attachment device that separates.Various embodiments of the invention also can be applicable to the cell of other type and separating of biomaterial, for example sperm, virus, bacterium, cell fragment, organelle, spheroidal structure, colloid outstanding body, cell body fragment and other biomaterial.

" particle " used herein is meant biology or other chemical material, include but not limited to low (gathering) nucleotides, many (gathering) nucleotides, chemical compound, protein, lipid, polysaccharide, ligand, cell, antibody, antigen, organelle, lipid, blastomere, cell aggregation, microorganism, peptide, cDNA, RNA etc.

The separating blood constituents illustrative methods comprises the steps: to provide the stream of first liquid with multiple blood constituent; Second liquid stream is provided; Contact so that first Disengagement zone to be provided with first liquid stream with second liquid stream; And divide opposite sex ground that second liquid stream is gone in the first haemocyte composition sedimentation of multiple blood constituent, remain on the second haemocyte composition of multiple blood constituent in first liquid stream simultaneously.Second liquid stream with first haemocyte composition then divides opposite sex ground to remove from first liquid stream with second haemocyte composition.

But it is the precipitation step speed band shape that the present invention is different or isopycnic.In addition, it is no turbulent flow basically that first liquid stream and second liquid flow, but and also laminar flow basically.

In optional embodiment, the first haemocyte composition is numerous red blood cells and numerous leucocytes, and the second haemocyte composition is numerous blood platelet.For the first haemocyte composition, can from numerous red blood cells, separate (passing through laser steering) outstanding many leucocytes in holographic ground.Other holography of the present invention is handled and to be comprised with holographic mode ground remove numerous pollutants from first liquid stream, isolates the biology fragment in holographic mode from first liquid stream, and isolates the second numerous haemocyte compositions with holographic aspect from first liquid stream.

Also can comprise in the additional separating step of illustrative methods: provide the 3rd liquid stream exemplary; Contact so that second Disengagement zone to be provided with first liquid stream with the 3rd liquid stream; And divide opposite sex ground that the 3rd liquid stream is gone in the second haemocyte composition sedimentation of multiple blood constituent, remain on the 3rd haemocyte composition of multiple blood constituent in first liquid stream simultaneously.But in embodiment, the second haemocyte composition is numerous blood platelet, and the 3rd haemocyte composition is a blood plasma.

The formation also capable of being combined of multiple separating step has multiple separating step and is connected in series, is connected in parallel or both the more labyrinth of combination.

The illustrative methods that fluid mixture separate is formed the composition that do not move about according to the present invention comprises: first liquid stream that is essentially laminar flow with fluid mixture is provided, this fluid mixture has multiple composition, and this multiple composition has the corresponding multiple rate of settling; Second liquid that is essentially laminar flow stream is provided; Contact so that first Disengagement zone to be provided with first liquid stream with second liquid stream, in the Disengagement zone, first liquid stream and second liquid flow essentially no turbulent flow interface; Dividing opposite sex ground from first liquid stream second liquid to be gone in the first composition sedimentation of multiple composition flows, with second liquid stream of formation enrichment and first liquid stream of dilution, remain on second composition of multiple composition in first liquid stream simultaneously, first composition has first rate of settling of the multiple rate of settling, and second composition has second rate of settling of the multiple rate of settling, and wherein first rate of settling is relatively big than second rate of settling; Divide opposite sex ground that the first liquid diffluence from dilution of second liquid stream of enrichment is removed; And in first liquid stream of dilution, handle second composition holographicly.

Second illustrative methods also can comprise additional separating step, and for example holographic the separation, it comprises: the 3rd liquid stream is provided; Contact so that second Disengagement zone to be provided with first liquid stream of dilution with the 3rd liquid stream; And holographic trapping second composition and remove second composition from the first liquid diffluence of dilution and join the 3rd liquid stream, remain on the 3rd composition of multiple composition in first liquid stream of dilution simultaneously.

At the embodiment of exemplary means that is used for fluid mixture is separated into the composition that do not move about of composition of the present invention, this device comprises: have first import that is used for first liquid stream and be used for second liquid stream second import first sieve passage; This first screening passage further has first outlet that is used for first liquid stream and second outlet that is used for second liquid stream, this first screening passage further has and keeps first liquid stream and second liquid to flow the parts of essentially no turbulent flow, the first screening channel adaptation is deposited into second liquid stream in first composition that allows multiple composition in first liquid stream in first liquid stream, with second liquid stream of formation enrichment and first liquid stream of dilution, and second composition of while multiple composition of maintenance in first liquid stream; The second optically transparent screening passage, it has first optics inlet and the outlet of first optics that is connected with first outlet that is used for first liquid stream, and the second optically transparent screening passage further has second optics inlet that is used for the 3rd liquid stream and second optics outlet that is used for the 3rd liquid stream; And sieving channel attached holographic optical traps with second optical clear, this holographic optical traps is suitable for producing holographic optical traps, to select and to enter the 3rd liquid stream from first liquid moving second composition that drifts.Separable heterogeneity for example can be that different blood separate or other biomaterial, isolates in the sperm that for example never moves about to move about.

Another device or the system of multiple composition comprises in the separation of the fluid: have first import that is used for first liquid stream and be used for the optical clear screening passage of second import of second liquid stream, this optical clear screening passage further has first outlet that is used for first liquid stream and is used for second of second liquid stream and exports; And sieve channel attached holographic optical traps system with optical clear, this holographic optical traps system is suitable for producing holographic optical traps, first composition of multiple composition entered second liquid stream during mobile first liquid flowed with selection with in first liquid stream, thereby form second liquid stream of enrichment and first liquid stream of dilution, and second composition in the multiple composition remains on simultaneously in first liquid stream.

Another embodiment that is provided for separating the various kinds of cell method comprises: the stream of first liquid with various kinds of cell is provided; Second liquid stream is provided; Contact so that first Disengagement zone to be provided with first liquid stream with second liquid stream; And divide opposite sex ground that first cell of various kinds of cell is moved adding second liquid stream, remain on second cell of various kinds of cell in first liquid stream simultaneously.This method also comprises usually: divide opposite sex ground to remove second liquid stream with first cell from first liquid stream with second cell.This method also comprises: in the 3rd liquid stream is provided; Contact with first liquid stream with the 3rd liquid stream, so that second Disengagement zone to be provided; And divide opposite sex ground that second cell settlement of various kinds of cell is gone into the 3rd liquid stream, remain on the 3rd cell of various kinds of cell in first liquid stream simultaneously.In addition, can holographic ground from the first liquid flow point from the second numerous cells, and holographicly from first liquid stream, remove multiple pollutant or biology fragment.

In another embodiment consistent, used as optics trapping (perhaps laser steering) technology of handling small multiplatform environments with the present invention.A kind of received description for this effect is that focused light (for example light that is focused on by high-NA microlens group) has intensity gradient jumpy closely.Optical trap uses the gradient force of light beam with the trapping particle, and to minimize its energy, the high particle of a permittivity ratio surrounding medium will move on to the optical trap zone based on its dielectric constant, and electric field is the strongest herein.

Optics trapping of the present invention be used to set forth the cell screening of carrying out with some modes and purify (such as, from pollutant (for example virus and bacterium)).For example, the power that is put on material by optical trap is responsive to the accurate distribution of dielectric constant, and this is because material-optical force depends on the composition and the shape of object.

Further, other power on the object is responsive for the shape of object and environment liquid fluid flow detecting material, size with as the hydrodynamic interaction between the control of the feature of surface roughness.

Further, anchored object allows other make visit automated method, for example high speed imaging and particle characteristics scatterometry in known location.

In an embodiment consistent with the present invention, in realizing many trap system, a diffraction optical element (" DOE " promptly utilizes the phase shift hologram of one of transmission or reflection geometry) is used to change the wave surface of single laser beam.This transformation wave surface is so that the laser beam in downstream becomes the single laser beam that its a large amount of relative positions and the direction of propagation are determined by the essence of diffraction optical element basically.

The invention provides by produce the optics of optical trap with the lens focussed laser beam, wherein lens have the numerical aperture less than 0.9, and preferred the minimizing up to optimization ground less than 0.1.

The screening that utilizes holographic laser to handle comprises formulates the recognition category that is used to sieve object, will sieve object and introduce the screening district, and utilize controlled laser to handle this object according to its recognition category.This operation can be based on its recognition category and carry out clamping, move, rotation, mark or use diverse ways to destroy this object.Thereby, the invention provides the parallel method of carrying out haemocyte screening and spermatoblast screening of holographic optics trapping of utilizing of carrying out.

In one embodiment of the invention, the spectroscopy of biological material specimens can be used and be suitable for stiff spectroscopy or the backscattered imaging light source of polarised light is finished, identification is useful for the evaluation chemicals for the former, and the latter is suitable for measuring the size of internal structure, for example examines size.Utilize this spectroscopic analysis, in certain embodiments, pair cell is inquired about.There is the spectrum of the cell of positive findings (that is the cell that reacts or combine with label) can utilize the imaging illumination to obtain.

Computer program can analysis spectral line the target wanted with identification of data (promptly carry the cell of X or Y chromosome, perhaps suspect it is cancer, precancerous and/or be not the cell type of cancer, or the like), can submit to information to guide phase diagram optical element (being optical trap) then to emanate or to comprise that want or selected target (being cell type).Involved cell can or combine based on the reaction of involved cell and chemicals and discern, perhaps utilize object natural fluorescence or with the fluorescence of this object association material as distinguishing mark or background indicia.After test is finished, can select by computer and/or operator, abandon which cell and collect which cell.

The operation of pair cell has multiple available light beam usually and is safer.Similar to nail matrix, multiple pincers have guaranteed at this intracellular any specific introducing energy still less.Eliminate heat spot like this and reduced hurtful danger.Any destructive pair of light quantum handled very useful, because absorb square in direct ratio with laser energy.Only increasing by second pincers has reduced in 1/4th of two light quantum absorptivities of specified point.The trapping maxicell has comprised a large amount of laser energies that are used for effective trapping.Making energy enter single trap can cause the moment of this cell is damaged.

Even only can obtain very big reinforcement by utilizing the holographic optics trapping to the operation of single cell.Can clamp by a team and operate single cell, this team clamps in a side along this cell of all long supports.The rotation that causes allows the observation of 360 degree to this cell.Except that this advantage of observation to biomaterial, also there is the ability of accurate directional sample, this is clearly for for example having the researchs such as scattering experiment of strong dependency very useful to sample orientation.

Adopt the screening in the wide visual field (wide field of view) many advantages to be arranged, for example higher output.Yet the standard pincers in a WFOV (the wide visual field) obtain can be owing to excessive radiation pressure is failed.There are the pincers that utilize the holographic optics trapping in the wide visual field to obtain to allow the ambient light pattern that forms radiation pressure that can very big minimizing light beam.Vortex trap is for example because the phase place that light changes disappears the center of trap and has the heart in the dark.This in the dark the heart mean that the great majority of the light of propagating along beam center no longer exist.This light beam contains great majority in the radiation pressure of light, and this fact is accurately, so the very big difficulty that has alleviated axial trapping of their removal.Other pattern, for example circular pattern has identical advantage.

In an embodiment consistent with the present invention, the method and system make himself have semi-automatic or automation to the moving process of following the trail of each optical trap and the processing procedure of content.In an embodiment consistent with the present invention, can monitor moving process by the optical data stream that can observe, perhaps be translated into vision signal, by operator's sense of vision, utilize spectrographic technique and/or observe by video-frequency monitor and it is monitored or studies.Optical data stream also can be handled with monitor intensity by photodetector, perhaps this optical data stream is converted to the numerical data stream that is applicable to computer and program by arbitrary proper device.The generation of the selected and optical trap of computer program control cell.

In other embodiment consistent with the present invention, track cells moves to be based on by encoding phase figure optical element and causes predetermined the moving of each optical trap.In addition, in certain embodiments, computer program has been preserved each cell record that is contained in each optical trap.

Thereby listed some feature consistent quite widely, in following detailed description, can understand preferably, also in order to understand preferably for the present technique document with the present invention.Certainly, there are some supplementary features consistent will be described below, and form appended accessory claim about this point with the present invention.

In this regard, describing in detail before at least one embodiment consistent, should understand this invention and not limit it and apply for description hereinafter or the structure of in description of drawings, setting forth and the arrangement details of assembly with the present invention.The method and apparatus consistent with the present invention can be used for other embodiment, and can actually in many ways apply and finish.And the summary that should understand here the wording used and term, comprises below is to be used to describe purpose and should not to be considered as limiting with ining addition.

Similarly, those of ordinary skills should recognize the disclosure based on notion can be easy to the design basis that is used to finish the some purpose structures of the present invention, method and system as other.Therefore, importantly, claims should be regarded as comprising this equivalent construction within the spirit and scope that do not deviate from the method and apparatus consistent with the present invention.

From following detailed description of the present invention and its embodiment, and accessory rights requires and appended accompanying drawing can clearer many other advantages of the present invention and feature.

Description of drawings

Purpose of the present invention, feature and advantage will become clearer with reference to disclosed content below in conjunction with the accompanying drawings simultaneously, wherein:

Figure (or " accompanying drawing ") the 1st is according to the side view of the device 100 of the embodiment of the invention.

Fig. 2 is the view that is used for the optics trapping of component separation in device 200.

Fig. 3 shows the schematic diagram according to two step section systems 300 of the closure that is used for the blood constituent separation of the embodiment of the invention.

Fig. 4 exemplarily illustrates the holographic optics trapping system according to the embodiment of the invention.

Fig. 5 is the schematic diagram of holographic optics trapping system that is used to sieve object according to the embodiment of the invention.

Fig. 6 shows (being divided into Fig. 6 A and Fig. 6 B) flow chart of a method embodiment according to the present invention.

Fig. 7 A and 7B are respectively schematic side elevation and the vertical views that is introduced into sample room according to a kind of sample of demonstrating of the embodiment of the invention.

Fig. 8 illustrates the scanning electron microscopy according to the sample room of the embodiment of the invention.

Fig. 9 illustrates the enlarged drawing according to the sample room workspace of the embodiment of the invention.

Figure 10 illustrates according to the example of the embodiment of the invention to the lateral deflection of screening.

Figure 11 A-11B illustrates front view and the side view according to the infundibulate trap of the embodiment of the invention respectively.

Figure 12 illustrates the cell sifter based on the rotation dish according to second and the 5th related application of the embodiment of the invention.

Figure 13 illustrates the optical peristalsis according to the embodiment of the invention.

Figure 14 illustrates the screening system according to the embodiment of the invention.

Figure 15 illustrates the screening system according to the embodiment of the invention.

Figure 16 is the side view according to the high-aspect-ratio plane sifter of the embodiment of the invention.

Figure 17 is the plane according to the high-aspect-ratio plane sifter of the embodiment of the invention.

Figure 18 is the perspective view according to the three-dimensional screening plant with multiple several sifters of the embodiment of the invention.

Figure 19 is the plane according to the multichannel sifter of the embodiment of the invention.

Figure 20 is the plane according to the sifter of the narrow waste liquid flow region of having of the embodiment of the invention.

Figure 21 is the plane according to the sifter of the different flow velocity of utilizing of the embodiment of the invention.

Figure 22 is the plane according to the sifter with several selector channels of the embodiment of the invention.

Figure 23 is the plane of the sifter of the screen area that narrows down according to having of the embodiment of the invention.

Figure 24 A is the side view according to the multilayer laminar flow sifter of the embodiment of the invention.

Figure 24 B is the multilayer laminar flow sifter plane according to the embodiment of the invention.

Figure 25 illustrates the result who utilizes various embodiments of the invention to carry out the screening of spermatozoon activity or movement.

Figure 26 is according to the exemplary screening of the embodiment of the invention and the block diagram of piece-rate system.

Figure 27 is according to the exemplary double reaction device product purification of the embodiment of the invention and the block diagram of piece-rate system.

Specific embodiments

Though the present invention allows many multi-form embodiment, as shown in the figure and specific embodiment that will at length narrate herein should be understood that the disclosure is a principle example of the present invention, rather than the present invention is limited to these concrete embodiment that sets forth.

As previously discussed, different embodiments of the invention are used for the composition of separating mixture, separate such as blood constituents different in the whole blood is separated into accordingly, such as blood platelet separate, red blood cell separates, leucocyte separates and blood plasma separates.These different embodiment utilize one to be or screening passage that many have the multistable laminar flow as described below, and allow one or more composition to divide opposite sex ground to be precipitated into another from a liquid stream, thus these component separation are become corresponding liquid stream.In addition, these different compositions can utilize optics machine machine (for example holographic optical traps) screening.Various embodiments of the invention are provided at the separation (for example in a continuous closed system) of carrying out composition in the continuous substrate (basis) thus, and the potential damage and the pollution that not do not cause especially for the art methods of blood constituent partition method.Continuous processing procedure of the present invention has also effectively been saved the time that blood separates.

Except whole blood screening with separates use, the present invention also is applicable to other cell screening application, such as the separation of carrying out cancer cell in marrow extract for example from normal or healthy cell.Various embodiments of the invention can further be applicable to other biology or medical domain, for example cell, sperm, virus, bacterium, organelle or subparticle (subparts), chondritic, the outstanding body of colloid, lipid and lipid ball, gel, immiscible particle, blastomere, the separation of cell aggregate, microorganism and other biomaterial.For example, according to the present invention, the separation of composition can comprise cell " cleaning ", therein pollutant (for example bacterium) is removed from cell suspending liquid, and it is particularly useful in medical science and food industry applications.It should be noted that based on the prior art of liquid stream and also do not recognize any application that utilizes the variable rate of settling and optical manipulation to carry out the screening of immovable cell component or separate.

Though following discussion concentrates on and sieves blood constituent and separate to form different blood, but device of the present invention, method and system can extend to the material of particulate, biology or the cell of other type, these materials can deposit or layering in fluid stream, or these materials can carry out optical manipulation between Different Fluid Flow.For example, method of the present invention can be used for separating spermatoblast that do not move about or non-activity from living cells, be deposited into second liquid stream by the cell that allows to move about from the first liquid stream, keep the swarm cell in first liquid stream simultaneously or allow swarm cell to move to the 3rd liquid stream.The sieve method of other cell separation also is feasible, for example realizes detached island cell, perhaps the island cell mass of separation different size from the pancreatic cell of other type by one of flow separation or optics light pincers (trap) or both.Use the present invention also can isolated viral, protein and other big molecules of the different rates of settling arranged.The holographic optical traps that uses together with different separation phase is useful to the cell or the separating of particle of other type particularly.

The present invention also has other medical application.For example, can utilize the part of following multiple laminar flow, remove the refuse in the whole blood herein in the reason and be back to patient as the kidney dialysis treatment.Therefore, except size separation, for example, the present invention can be used for separation based on diffusion, motility and other gradient type based on relative density.

For example, the present invention can be used for moving composition from a kind of solution to another kind of solution, is useless or does not expect and separate by filter or centrifugal action at this.These application except above argumentation, other application comprises the colloid (being used for research or commercial the application) of segregation given size from the colloid of other size, and clean particle (for example cell, egg cell, or the like (substitute effectively they are contained in medium wherein and clear the pollution off), or from the solution of salt and surfactant with the solution cleaning particle (for example nanotubes) of different salinity or surfactant-free.

The activity of these separated components can be depending on many physical propertys of object, comprises autokinesis, self-diffusion, free-falling speed or depends on activity under the external force effect (for example electromagnetic field or holographic optical traps).Available these characteristics that it sieves can comprise that cell mobility, the anti-viability of cell, object size, object quality, object densities, particle attract or repel the trend of another or other object, image charge, object surface chemical property and some molecule are adhered to the trend of object in liquid stream.

Though the present invention has at length discussed about installing 100,200 and 300, be construed as these argumentations and be equally applicable to other different embodiment of in Figure 13-24 and 26-27, illustrating.

Fig. 1 is the side view according to device 100 of the present invention.Routine as shown in Figure 1, screening plant 100 comprises screening passage 110, a plurality of import 120 and a plurality of outlet 130.Corresponding fluid stream (for example shown liquid stream W, X, Y, Z) enter one of them import 120 and by these screening passage (or screen area) 110 essentially no turbulent flows or as laminar flow flow through and by exporting 130 outflows accordingly, as shown in the figure.

Device 100 (with following 200) can make or utilize body material miscellaneous to form the element that separates by multiple material (for example metal, pottery, glass and plastics).Different materials and manufacture method are discussed in third and fourth related application, and comprise and for example use the different condensates that ultraviolet ray is handled down that are exposed to.Other details also provide in third and fourth related application, for example using and selecting, for example combination of syringe pump, peristaltic pump suction, weight-driven suction and different suction movement dissimilar aspirators.

In selected embodiment, when the optics light with holographic trap or other form clamped logotype, device 100 (or 200) were transparent for the holographic generator of selected wavelength, are optically transparent when holographic generator utilizes visible wavelength for example.Depend on selected application, device 100 (200) also should be that process is sterilized and also have the temperature of control.Can different fluid stream be injected import 120 by the known variety of way of those of ordinary skills, comprise within the scope of the invention and use peristaltic pump or gravity to inject, and this method also can be used for controlling different liquid stream W, the flow velocity of Y and Z.When using peristaltic pump,, the bubble trap can be combined in import 120 places of device 100 (or 200) in order to keep constant flow rate and pressure.

The different liquid streams that separation of the fluid utilized can be diluted or concentrated, with volume that increases or reduce a kind of solution wherein or the concentration that influences some dissolving or suspension material, or influences the physical characteristic of solution, for example viscosity, temperature or density.When being used to the blood screening, a sample of device 100 comprises: (a) dilute blood, to reduce the hydrodynamical interaction between obstruction and the haemocyte, (b) expanded in volume that separates of blood or blood, (c) blood, blood separate or influence the correction of density of the another kind of solution of fluid behaviour or screening behavior, (d) under the condition that the expansion of solution capacity is removed from system with the increase, particularly fluid volume of the volume that keeps fluid.As following more detailed narration, different chemical inhibitors or protective agent can add to liquid stream, and these materials also can have different temperature and levels of viscosity to improve the seminal fluid screening.

The different liquid streams that fluid is utilized in separating can be " activated ", mix by some process in some external effect activation solution, or with external solution.The sample of external effect comprises: (a) be added electric field, (b) apply magnetic field, (c) be exposed to light, (d) revise temperature, (e) introduce chemical substance, (f) introduce biomaterial, (g) shear solution (shearing thesolution) reaches (h) vibration solution.The activation that is caused by external solution comprises: (a) arrangement of particle, molecule or cell, (b) polarization of one or more composition, (c) crosslinked, (d) beginning of chemical reaction or termination, (e) beginning of biologically or termination, (f) chemistry, physics or the type of biologically or the change of speed, or (g) cause reaction according to the characteristic of the special composition of reacting or separate.The sample that is used for the device 100 of blood screening comprises: (a) add reagent, condense with minimizing; (b) add reagent to keep the activity or the health of blood solution or its composition; (c) add such reagent, thereby this reagent can by for example bonding or assemble near certain composition physical property of influencing one or several enlarge screening process, comprise that interpolation bead or other can attach to the particle or the polymer of one or more composition, comprise that also salt or other can influence the material of electrostatic interaction or the fluid dynamic size or the material behavior of material; (d) add the reagent that for example influences fluid behaviour by change density, viscosity, surface tension or other parameter; (e) add the reagent that strengthens or suppress certain material gathering; And (f) add to strengthen or suppress the reagent that certain material attaches to other material or flow device parts.

One of them fluid stream according to the present invention (the liquid stream W for example) comprises multiple composition A, B, C and E.For example, when fluid was whole blood, these compositions can be red blood cell (" RBC "), and leucocyte (" WBC "), blood platelet, cell debris and pollutant are all in blood plasma.Typically, the majority in numerous compositions has the different rates of settling, typically utilizes the svedberg coefficient to measure.For example, red blood cell has the rate of settling bigger than blood platelet, and will be in the substrate that plays the support function effect deposition velocity faster, belong to since gravity or buoyancy, do not have other mechanism of action, for example intervention of centrifugal action.Flow through screen area 110 when different liquid flows W, X, Y and Z, based on the different rates of settling, multiple composition (for example cell or other particle) will deposit, and drift moving thereby flow to another liquid from a liquid.Go out as shown, composition A with relative maximum settlement speed is illustrated from liquid stream W and moves to lowermost layer liquid stream Z, composition B with relatively secondly high rate of settling is depicted as from liquid stream W and moves to liquid stream Y (on Z), composition C with smaller rate of settling is depicted as from liquid stream W and moves to liquid stream X (on Y), and has the composition E that compares the comparatively minimum rate of settling to be depicted as still in liquid stream W (on Y).Utilize different settling characters, each of separable these compositions enters corresponding liquid stream, and isolates mutually and flow out from corresponding outlet 130.Because every kind of liquid stream W, X, Y and Z flow out by its corresponding outlet 130, this kind liquid flow point opposite sex ground is removed from other liquid stream, and promptly a kind of liquid stream has been removed and another kind of liquid stream keeps motionless fully or respectively other is removed from remaining liquid stream.In addition, this differentiation formula is removed and can be walked abreast, and just all liquid streams are removed simultaneously or continuously.

Continuation utilizes the whole blood that adds anti-coagulants (for example natrium citricum or heparin) to flow W as fluid, for example with reference to Fig. 1, different blood separates can be separated from one another, because the red blood cell deposition is the fastest, (for example represents with composition A, 4.59 μ m/s) and leucocyte with slow falling slightly long-pending deposited at rates, represent (for example, 2.28 μ m/s) with composition B, blood platelet is with relatively slow deposited at rates, (for example represent with composition C, 0.055 μ m/s), and blood plasma still constitutes liquid stream W, E represents by composition.Each blood separates and can remove by corresponding outlet 130.

Do not have among Fig. 1 separately expression, because buoyancy and relative density, have the liquid stream (for example emulsion) that particle or composition will upwards flow to higher position among one or more in fluid stream.For example, the little particle of density that flows into liquid stream X can rise up into liquid stream W, flows out with liquid stream W by corresponding outlet 130.

Shown as side view, device 11 screening passage (or screen area) 110 has length variable " L " being parallel to flow direction, have the variable degree of depth " D " perpendicular to flow direction, and the shown width " WW " (WW narration like this is for fear of obscuring with liquid stream W) that extends in the page.Select these different sizes based on several factors, particularly the flow velocity and the rate of settling of interested composition.For example, because selected flow velocity, the total length of screening passage should be sufficiently long with the composition of minute opposite sex ground mobile phase to the slowest rate of settling, composition C as described in Figure 2, simultaneously in the shorter accordingly length of other liquid stream with separate and subside speed composition faster, the liquid stream Z that contains composition A as shown in the figure and contain the liquid stream Y of composition B.

The aspect that the present invention further is different from prior art is that quite bigger tolerance or permission are arranged on length-width ratio, but still to have kept be laminar flow basically.Length for the Aspect Ratio of width as can be 5 (or bigger) than 1 (5: 1), (this moment, length was greater than width) to about 1 (perhaps littler) than 2 (length is less than width at this moment) variation.The length-width ratio of one preferred length and width is approximately 2: 1, and can change from 3: 1 to 1: 2.

Flow velocity also can change between a large amount of liquid streams that device 100 is utilized.For example, lower level liquid stream (for example Y and Z) higher flow velocity can tend to compress liquid stream W and X, causes in short distance more special component must cross deposit and enters liquid stream Y and Z.

In addition, are speed band shapes typically by the deposition of installing 100 different liquid stream compositions, promptly separates with size two aspects, in addition the shape and the static characteristic of material based on the relative density of this composition.Yet, under other condition, for example low flow velocity, thin flow depth and/or long screening path 10 0, but also isopycnic of deposition, that is, only based on the relative density of composition.

When wishing that isopycnic is separated, comprise that the different liquid streams of liquid stream W, X, Y and Z can be used for selected the separation to be equipped with mutually with composition density through selecting and calibrating to form the density gradient of expectation.For example, the different liquid streams that comprise liquid stream W, X, Y and Z through selection with calibration so that a different density cumulative or decrescence respectively to be arranged, thereby form the gradient density gradient, different particle depositions is in suitable step.In addition, by utilizing abundant fluid fluid layer, the density step effectively becomes continuous, and corresponding Crystallization Separation ability is arranged simultaneously.

Comprise the laminar flow different fluid of liquid stream W, X, Y and Z for example roughly, also can be selected for the component separation of specific (special) requirements according to suitable standard.For example be used for blood and separate, different liquid streams are formed whole blood, and for example liquid flows W, and blood plasma or cushioning liquid flow as remaining liquid.This different fluid is process preliminary treatment before entering by import 120 also, for example by dilution, the composition additive (anticoagulant for example for example that adds other, flocculant or cementing agent), the preliminary treatment of other isolation technics is operated or passed through to viscosity or other fluid speciality.Again for example, whole blood can preliminary treatment tentatively to remove some red blood cells or to add anticoagulant (for example natrium citricum).

Though illustrate with four liquid stream or passage, being construed as device 100 (or following 200) can be with many liquid streams and corresponding liquid inflow entrance 120 and outlet 130 enforcement.The sum of a fluid flow be restricted to according to have the ability to keep each liquid stream laminar flow basically or the non-turbulent flow state.So,, any unwanted liquid stream minimizes so that mixing so that each interface between the liquid stream is non-turbulence state basically.In addition, also has the consideration of relative density aspect, because fluid comprises the liquid stream that utilizes the regulation separating step that also can cause the restriction of liquid flow amount.

Different device 100 (or following 200) can be further cooperates with additional device 100 (200, below), parallelly is used for higher flow and series connection is used to increase separation phase, for example in order to improve the purification level.In addition, different devices 100 also can separate combination with nondeposition, or cooperate with subsequently the separate series of utilizing nondeposition machinery, with with composition line bonus between the liquid the finished stream from cooperating, for example, utilize optical force, for example the holographic optical traps in the 5th related application is quoted herein with for referencial use.And different devices 100,200 or 300 can have different size and many different passages or liquid stream.

Fig. 2 (and Figure 13) is the bulking property explanation that utilizes the optical force that such holography that is separated by the supplementary elements that are used for carrying out at device 200 or optical trap produce.To the formation of a large amount of holographic optical traps 210 with operate in below with reference to Figure 4 and 5 more detailed description is arranged, wherein install the sample 506 of 200 pie graphs 5.Two kinds of liquid stream W and X are shown in Figure 2, and liquid stream W has two kinds of composition A and B at first.Utilize holographic optical traps 210 (in Fig. 2, being described as conical part) thus catch " A " composition, and move them and enter liquid stream X.Such optical trap is useful to increasing the special purity that separates particularly, particularly to having separating of insufficient difference based on the rate of settling.Such optical trap is also particularly useful for removing undesirable composition, for example cell debris and other impurity.In addition, to the mixing of composition or mix and can realize in process at above-mentioned speed belt course flow point, optical trap is accurate especially when removing undesirable composition.For example leukocytic smaller part may sedimentation be not enough soon, cause in blood platelet separates, having some leucocyte pollutants.Can utilize optical trap to select and remove leucocyte and enter independent liquid stream, thereby increase the purity that blood platelet separates.

When being used for blood screening application, should be appreciated that, be better than leukocytic blood platelet and erythrocytic optical manipulation (or " pincers obtain ").But utilize low numerical aperture in system 500 (below argumentation is arranged) to improve significantly leukocytic optical manipulation.

When using execution simultaneously with optical trap, device 100,200 or 300 should utilize for the optically transparent material of selected optical wavelength and implement.When holographic trap is that the corresponding transparent material that can utilize other is to be suitable for selected wavelength when other wavelength is carried out.Carry out and place this device 100,200 or 300 being labeled as the position of sample 506, and utilize system 500 to carry out this holographic optical traps, as one of a separating step of the present invention or a part at Fig. 5.

Fig. 3 shows the two-stage system 300 according to the closure that is used for blood or other component separation of the present invention.In the phase I 305, flow into by import 315 from the blood constituent of selected donor and to form first liquid stream, and blood plasma returns (or being preferably in initial starting point) by import 320 and forms second liquid stream.First and second liquid stream is not turbulent flow, in other words is laminar flow, and forms non-turbulent contact each other in first Disengagement zone 325, forms non-turbulent junctional area between two liquid stream.In first Disengagement zone 325, red blood cell and leucocyte both are deposited into second liquid stream from the first liquid stream, and are to concentrate in the reservoir 330, for other application (for example as medically being used to assemble cell) or be used to return selected donor.As noted above, the flow velocity of the length of first Disengagement zone 325 and first liquid stream is scheduled to, so that red blood cell and leucocyte all have adequate time passive deposition under gravity and buoyancy to become second liquid stream.

Continuation is with reference to Fig. 3, and having removed wherein now basically, the continuous path of flowing through of red blood cell and the no turbulent flow of leukocytic first liquid stream enters second separating step 310.In second separating step 310, first liquid stream enters second Disengagement zone 340 with the 3rd liquid stream from import 335.In this exemplary embodiment, the 3rd liquid stream is also to be made up of the blood plasma that comes from selected donor.In second Disengagement zone 340, remain in blood platelet in the liquid stream and deposit passively and enter liquid stream 3, and in reservoir 345, assembles, to be used for the medical science use, for example with the blood plasma of liquid stream 3.The liquid stream of further removing 1 is to return import 320 and 335, each self-forming first and the 3rd liquid stream from exporting 350 outflow back recirculation.Point out as previously discussed, inject donor blood plasma in for example a part when system 300 can system starts, or (be basically or mainly be blood plasma) exporting the generation of 350 places to flow 1 with another biocompatible nontoxic liquid stream up to the liquid of removing when initial by centrifugal selected donor blood.

The separation that yet can utilize auxiliary these blood of additional holographic trapping separating step to separate is not shown separately in Fig. 3.For example, can utilize holographic trapping separating step instead or append to second stage 310.In addition, illustrated that second step is separated to have utilized liquid stream 1 that in other embodiments, liquid stream 2 can separate through second (or more) step, in addition or replace liquid stream 1 additional separating step.And additional separation phase can utilize series connection or in parallel.

More generally, the Disengagement zone is that wherein material sieves or separate areas partially or completely based on some material behavior.Can be single in any different liquid stream in embodiments of the present invention of Disengagement zone, in turn, or the one or more following technology of simultaneous use:

(a) separation of rate of settling band shape: separate by the rate of settling.The rate of settling mainly is the function of material size and density, also is the function of material shape and static characteristic.For this separation, set up laminar flow and under the gravity effect, or the inertia force (for example revolving force of a centrifuge formula device) by other separates under some situation.

(b) pass through density separation.Separate for this, set up linear, step, step or the density gradient that replaces and allow material in this gradient, deposit and/or emulsification arrives or is in the zone of mating the density environment near arriving this material up to material stably.

(c) diffusivity is separated: separate by diffusivity.Separate for this, material is based on the distance of their diffusions in the regular hour and separates.

(d) motility is separated: separate by motility.In this separates, material based on material in the regular hour from the motility that himself has and mobile distance and separating.

(e) optical fractionation: utilize optical force to separate, typically need not reception and registration of feedback mechanism ground and influence based on the optical system that individual subject is analyzed.

(f) direct optical fractionation: utilize optical force to separate, typically utilize feedback mechanism to pass on and to influence based on the optical system that the individual subject analysis is carried out.

(g) dielectrophoresis separates: utilize dielectrophoresis to separate.This power relies on the position of object in the electric field that is applied to put on this object, and insulator is reacted according to material and its environment.

(h) electrophoretic separation: utilize electrophoretic separation.This power rely on object in the electric field that is applied the position and the electric charge of material and its environment put on this object.

(i) magnetic separation method: utilize magnetic separation, this power puts on object according to its position in adding magnetic field and the magnetic of object.

(j) surface tension is separated: utilize the surface chemistry of surface tension or material to separate.This can for example relate to attracteding to or the structure of the liquid stream interface of certain material of stable existence.

More specifically, being used for blood screening demonstration partition method comprises: (a) utilize the separation of rate of settling band shape, separate one or more haemocyte types from some or all of other blood cell types and/or from blood plasma or broth; (b) as (a), separate, but utilize isopycnic to separate; (c) utilize deposition only from solution, to remove red blood cell (RBC) or red blood cell and leucocyte (WBC); (d) utilize deposition Platelet Concentrate from blood plasma; (e) utilize dielectrophoresis, electrophoresis or magnetic separation to extract red blood cell; (f) utilize optical technology to comprise that the optics pincers obtain and optical fractionation concentrated or extraction blood platelet from solution; And (g) utilize and can combine (for example particle of functionalization) reagent with the special cells type and according to any effect separating blood constituents of the above isolation technics, after that, reagent from or not from this cell type releasing.

For the blood screening, combined method may be the most effective, and for example: at first utilize the most of RBC of separation and Extraction and the WBC of rate of settling band shape, the blood platelet that utilizes optical fractionation to extract and concentrate then is discussed below.The optical fractionation step will not only be used for Platelet Concentrate, and (this method is also passable, for example, take in the centrifugal action step at last), and second step is provided, to bring into play strong inhibitory action to assembling jointly of WBC chance, be used for the example of blood platelet aphoresis with blood platelet.Such concentration step also can comprise filtration, for example separates or blood plasma filters WBC separating from blood platelet.

Though described device 300 about the blood partition method, it will be understood by those skilled in the art that except such blood partition method, device 300 can be used for separation miscellaneous.In addition, can think that also device 300 is specific embodiments of serial associated separation steps of the present invention.

Cell " cleaning " also is device 100200 or 300 1 effectively application.This cleaning can comprise the variation of medium, is used to store, preserve or other goals of medicine.This cleaning also can comprise by separating the interested cell of institute and enters another dielectric fluid that does not contain such pollutant and flow, and comprises for example medium of bacterium of pollutant thereby remove one.As previously discussed, the sperm separation also is an effectively application of device 100,200 or 300.

Screening plant 100,200 or 300 part or can observe with optical instrument from output wherein.This can for example use a camera for utilizing the directly visible imaging of directly natural daylight (brightlight) imaging or fluorescence imaging.Perhaps, can be for example dynamics light scattering or the dilatation wave spectroscopy of spectroscopy, propagates light spectroscopy, frequency spectrum imaging or scattering for example of complicated technology more.In many cases, these viewing areas can directly be integrated with liquid stream device to characterize input, output or intermediate steps.They can be used for diagnosis or hold the record, and perhaps can be used to pass on this overall process, and for example feedback processing is speed that how to finish or liquid stream or the amount of using each solution.In some cases, the optical observation zone can be used with additive, this additive for example in conjunction with or influence the chemicals of the part of solution, or the particle of fluorescence is solidified and/or sent to functionalization when certain material or disease exist.For the example of blood screening, these technology can be used to measure cell concentration, with the detection disease, or in detecting other parameter that characterizes blood.

Screening plant 100,200 or 300 some parts or can electricity consumption learn to do segment table from wherein output and levy.For example, screening plant can be implanted electronic device.For example, electronic device can comprise: (a) electric capacity, (b) galvanometer (c) is used for determining the ohmer of the volume conductance of fluid, can measure concentration or composition by this parameter, or (d) PH measuring instrument.Screening is used for blood, and the measurement of cell concentration degree, iron content, flow velocity, TCS, concentration of electrolyte, pH value and other parameter can be the screening plant part and parcel.

Screening plant 100,200 or 300 liquid stream composition can be passive, thereby are undertaken from control fully externally by the flow velocity of input and output.Perhaps, can in (not being described separately) device, implant the liquid stream composition work, the valve that for example can utilize electronics, optics, calorifics, mechanics or other influence opens or closes partially or completely.Screening is used for blood, and screening plant 100,200 or 300 can be useful on screening and/or transmit the integrated approach of one or more solution.For example, the side that consumable screening plant is made at reservoir has deformable film.This reservoir can be filled with solution, and for example a kind of and patient are biocompatible and can be used for the buffer of dilute blood.Another example is that it can fill anti-agglomerating agent.Carry and/or the utilization of a this fluid can be oppressed this film and carry this fluid dynamically by the influence control of machinery.Perhaps, can transmit this fluid with another mechanism.For special purpose, for simplicity and save, in screening process, need the set of different fluid solutions in different step.But merge the wastage in bulk or weight of simplification and the minimizing ownership and the operation of these composition certain degrees.It also can reduce the risk of pollution and error.This reservoir that holds input and buffering solution or fluid for example has been shown in Figure 14 and 15.

Device 100,200 or 300 can comprise the aspect (not illustrating separately) that one or more kinds of fluids or fluid composition are carried out biology or chemical analysis.This can comprise the measurement of pH value, the interpolation of particular biological or chemical material, or other measurement.Use for blood screening, this can comprise disease detection, different cell types or material are characterized in the concentration of blood plasma, the determining or other the estimation of blood quality, type or category of the sign of iron content, blood group.

Device 100,200 or 300 can comprise a zone (not illustrating separately), and soln using optical method (for example being exposed to ultraviolet lamp) or other method are carried out sterilization.Sterilization can to this install initial part or add to wherein be used to cushion, clean, dilute or other the solution effects that influences sample solution.Perhaps, sterilization can align the part or all of effect of processed sample solution.Screening is used for blood, and the solution optics sterilization of using for this device except that whole blood is important.Also have, the sterilization that blood or certain separate also is important.

This device 100,200 or 300 can comprise that the material finished has been constructed on one or more surface so that carry out the reaction of physics or chemistry with certain material.For example, surface can functionalization in case certain material adhesion in it, in order from this solution, to extract these materials or for diagnostic purpose.Screening is used for blood, and the surface of functionalization can be used for extracting unwanted material from certain separates.Optionally, they can be used for assembling the material of low concentration to be used to measure the concentration of an existing material or a kind of types of material, for example disease detection.

Screening plant 100,200 or 300 can comprise and wherein sieves concurrently but do not have the zone of physics wall separating liquid stream.For example, can there be a plurality of imports 120 and outlet 130 in the zone of parallel screening, and some are wherein arranged is similar each other on the function.Be used for replacement and be different from multiple parallel sifter physics current divider, shunting realizes as the result that physical property caused of solution and liquid stream.The available simpler and cheap device in parallelization sifter zone of these contacts produces high screen rate.They also can carry out to avoid the contact on surface.For the blood screening, available these zones minimize contact area and maximize screen rate and reduce consumption and complexity simultaneously.

But design apparatus 100,200 or 300 is used to control the zone (not illustrating separately) of flow rate or flow performance.For example, need laminar flow in many cases, often also wish to obtain special flow profile.In other cases, should there be consistent flow rate and behavior in several zones of device.Since this reason, the regional effect flow behavior that often need have shape and other characteristic.In some cases, this is finished by very symmetrical flow design.In other cases, the change that the flow region diameter is big and/or exist reservoir help to keep the unanimity of flow rate.In other cases, well-designed passage provides the accurate balance to required flow rate.In order to keep laminar flow, it is important that channel size has the zone of slow variation.Barrier or current divider also can be used to keep laminar flow.For the blood screening, the adjustment of flowing is important to reach high separation rate in output that keeps separating and purity.

Do not illustrate separately, device 100,200 or 300 can contain the mechanical mixture zone that design is used for liquid stream, for example excites the zone of turbulent flow.For example, flow to can turbulization and mixing into a large scale zone for liquid that quick narrow zone arranged.For the blood screening, a Mixed Zone can be mixed dilution or anticoagulant with blood, or as required other solution is mixed.

Device 100,200 or 300 can contain design and be used for the zone (not illustrating separately) of arranging cells someways or raw material.This sometimes finishes by shear flow, but also can by apply the outfield for example electric field finish.

Device 100,200 or 300 can contain the zone (not illustrating separately) that design is used for dissolved cell or decomposes raw material.This can be by shear flow, vibrate, force pass a hole, electricity or other device finish.For the blood screening, this may be valuable for the condensate of removing particular cell types or generation.This also may be to diagnostic purpose, for example disease detection or to belong to the parameter measurement of cell inclusion valuable.

This device can contain the zone (not illustrating separately) that makes cell expansion or dehydration or object, for example by introducing the reagent that changes osmotic pressure or passing through to change physical pressure.This can, for example, finish before with the density of adjusting cell or object carrying out isodensity screening as a step.It also can be finished and be used for killing or hitting special component.With the blood screening is example, and this can be used as the after-stage purification step to remove or to control undesired one-tenth and assign to finish.

The zone (not illustrating separately) that device 100,200 or 300 can contain heating or cool off one or more solution.This can realize the influence to its physical attribute, for example viscosity and density.Perhaps, it can realize the influence to its chemical attribute, for example chemical reaction rate or chemical stability.Perhaps, it can realize its influence to biological attribute, and is for example active, metabolic rate, or viability.For example, a kind of fluid stream can cause the movable sperm of energy to flow to the liquid stream that reaches cold away from the liquid of heat than the temperature height of another fluid stream.Also have, it can be finished realizing system-level compatibility, for example preparation of carrying out for subsequent processing steps.With the blood screening is example, and the solution that is used to be back to patient remains on suitable temperature to avoid that patient is felt cold.The solution of storing then can cool off in operation to keep those compositions or to be next step operation or to store step and prepare.

Device 100,200 or 300 can comprise the solution that some reservoirs are used in carrying out with storage process, or the solution (not illustrating separately) of generation in process is carried out.Having the reservoir that these and screening plant combine has simplified operative installations and has reduced required extention.With the blood screening is example, and reservoir can be equipped with anticoagulant, required any other solution in dilution and the process.Reservoir can also merge to preserve separating and discarded separating of sieving.

Device 100,200 or 300 can comprise the zone (not illustrating separately) that strengthens degree of mixing by diffusion.For example, mix when two strands of laminar flows contact, make it parallelly be divided into many narrowly, the liquid that contacts with each other stream has strengthened whole composite rate by diffusion.With the blood screening is example, can be used to spread Mixed Zone mixed diluting liquid, and anticoagulant or other contain the solution that whole blood or blood separate.

Do not illustrate separately equally, device 100,200 or 300 can include the zone of bubble trap with the air bubble in the removal system.This can make air bubble finish from the zone that flow region rises on the original flow region by one.With the blood screening is example, and this can realize can not passing on patient or the collection sample with the little air bubble of assurance from system loading or operation phase in a simple manner.

As implied above, device 100,200 or 300 can comprise some zones (not illustrating separately) that are used for suppressing the vibrations (pulsation) in any liquid stream, and for example those produce when peristaltic pump uses.A kind of method of vibration-inhibition is to integrate one " bubble trap " in device.The existence of an air bag that contacts with fluid allows the increase and decrease of air bag with compressed fluid pressure.Thereby air bag plays the effect of a damper, makes to flow smoothly.Other device can use too, and a slice elastic membrane for example can be crooked under higher pressure, thus smoothed pressure and flow rate.For the application of blood screening, the shock attenuation zone will produce more accurate and level and smooth flowing, and therefore higher screen rate, purity and output are arranged.

Device 100,200 or 300 can comprise the zone (not illustrating separately) of display unit state.For a kind of consumptive material, whether it can sterilize by indicating equipment, or does not use.For the application of blood screening, people will want to confirm that by an indicator screening plant sterilized and be not used or pollute.

Screening plant 100,200 or 300, or whole screening system can comprise one group of mechanism and be used for accurate fluid sifter level.This is important, because for some sifter, is not under the situation of accurate level at device, and buoyancy can cause mobile unintentionally and screening output and purity are produced negative influence.In addition, but screening plant can comprise whether balance or depart from the element of the balance number of degrees very of display unit.For example, it an electronics can be arranged or be incorporated on self sifter based on the balance of gravity.A passage that example is a moulding of this device, in fluid and an air bubble are arranged.But the angle that the position display unit of air bubble tilts.Another kind of such device shows the inclination angle with a Metal Ball that is arranged in slide rail.Another kind takes the form of uses optical correction, for example allows light source come or allow light source pass a wedge to determine its direction from a return reflection surface.For the application of blood screening, level control and indication are of great importance to guaranteeing high yield and high purity product.

Screening plant 100,200 or 300, or whole screening system can comprise the uniform temperature that is used for holding device and/or solution and/or constant mechanism's (not illustrating separately).This may cause and be not intended to flow and be important to the thermic buoyancy that screening output and purity produce negative influence eliminating.In addition, screening plant can include indicator, or does as a wholely in screening system, and indicator is indicated the temperature and/or the temperature uniformity coefficient of one or more compositions.For the application of blood screening, the uniformity coefficient of control and indicated temperature can be of great importance to obtaining high yield and high purity product.

Screening plant 100,200 or 300, or whole screening system can comprise and measure the one or more horizontal plane of solution and mechanisms of concentration (not illustrating separately) of inputing or outputing.With these measured values can decision operation speed, deadline, error condition, comprehensively monitoring or other application.For the application of blood screening, can when collect enough samples with horizontal plane and intensity indicator indication, or detect when solution makes a mistake or turns.

At last, screening plant 100,200 or 300, or whole screening system can have the inversion filling of the standard utilized or drain method, with the method for one or more fluid start-up systems.Cleaning can be finished simply by draining this device or crossing wherein to come with flow of solution.Screening carry out early stage, start solution can up to it by most of dilution and when having obtained required solution just be dropped.Similarly, in the later stage that screening is carried out, available a kind of fluid comes the material that sieved is discharged system from the beginning to the end to obtain minimum waste and maximum output.

Different screening plants 100,200 or 300 also can be provided the system of the device of common purposes to utilize by a kind of, and this device allows user to extract in the solution one or more to separate, and it depends on the scope of S value (size, density, or big or small * density).Figure 26 is the structure chart of exemplify illustrative screening and a piece-rate system 2600 according to an embodiment consistent with the present invention.The control (user imports 2610) that the sifter of this common purposes can respond the user changes the flow rate of each I/O channel.Customer controller can allow the user to determine scope by the S value of each output channel.This device will be a valuable laboratory tool, be used to implement the diagnostic assessment of various purifications, washing, separation and sample.It also will become the important platform that a research, exploitation and original shape are used.

The hardware of screening usefulness can be included in the shell, has and maybe may not have temperature control.This hardware comprises following part: (1) consumptive flow plate 2620 has several inputs and output on (for example screening plant 100,200 or 300), (2) several can be by computer-controlled peristaltic pump 2625, (3) temperature control and monitoring arrangement 2630, (4) reservoir support (or reservoir 2635 and 2640).Screening is by computer 2650 control, and this computer also monitors and control flow rate, temperature and other in order to control or diagnostic purpose and additional part.

Flow plate 2620 can be a simple purposes device usually, is suitable for aforesaid multiple application.It can contain two to four inputs and two to ten outputs.It can with sterilize, preprepared, the sealing form provide.Those reservoirs can be the parts of flow plate, or are attached to the part of sterilizing separately before use of flow plate.The all or part of cover glass that can cover of central authorities' screen area is to allow to carry out the operation that sample was killed/cut to light pincers, photoelectrophoresis or laser.

2650 pairs of pumps of computer and temperature control hardware are measured and are controlled.It also can intervene the hardware component that other can be coupled with.User software provides one before to bring in all pumps of real-time control very easily.It can carry out necessary calculating and control the speed of each pump to give the user specific desired the flowing of screening application.

System 2600 can be very general in to allow multiple screening to use, and perhaps independent running is perhaps cooperated with an additional laser equipment.These application include but not limited to: recognize cell, shine cell killing, usefulness light electrophoretic separation cell, the swarm cell separation based on motility, the passive screening object of usefulness diffusion method with high intensity laser beam with fluorescence method, and liquid flows the passive screening of banded density.Liquid flows the passive screening of banded density and is widely used, usually can be by minimum or do not have supplementary equipment therefore and finish.

The screening of passivity Flow Control and/or the use of optics step power (as described below) have been used in exemplary use and the application that is used for the general purpose screening system 2600 of this kind, list in the form below:

Kind	Summary	Details
			Clean removal type composition	Be used to remove the cell cleaning of disease	From people's seminal fluid, remove dead cell, bacterium and virus and from animal semen, remove dead cell, bacterium and virus
Condensate is removed	In industrial process, from material, remove the coarse drop in the aggregation colloid removal emulsus agent in the undesired material grumeleuse removal solution
		Precursor is removed		Removing precursor material from interrupt to cultivate operation separates cell that differentiation is arranged remove blastocyte from the sperm cultivated and ovum
Cleaning-change medium	Cell is handled				Changing infiltration condition---painted automatic multistep test cell line of cell expansion or contractive cell or operation suppress active sperm fluorination reaction
		Cell is handled	Particle death
	Chemical examination			Target solution reaction chemical examination
		Diagnosis	Characterize the solution of monodispersity		The composition of the density measure particle of the dimensional measurement known materials particle of S (Svedberg coefficient) the measurement known materials particle of measurement particle
Characterize the solution of polydispersity	Measure the distribution of S, size, density, composition
			Disease detection	The sample that extracts bacterium from infected tissue extracts bacterium from the stool sample sample extracts bacterium from blood sample extracts the cell of virus infections from healthy cell

	Environment	Extracting spore from sample extracts particulate and is used for environmental monitoring and is drinking or swimming water monitoring water security extracts from environmental samples and propagates thing quantitatively to determine the mould level the infection family to determine airborne spore quantity from water or air
				Food security	From food, extract bacteria sample and be used for diagnosis
Purification-minimizing impurity	Virus	The purified virus sample
			Purification-extraction composition	Cell component	From the solution of dissolved cell, separate different cell components
Cell component	The isolated cell device
		Bacterium		From infected tissue, extract bacteria sample
Spore	From solution, extract the spore sample
		Filter		From food, filter big composition	The filtration yeast filters yeast and filter fat from milk from liquor from beer
Industry is filtered	From water, remove particulate and from machine oil, remove particulate
			Medicine filters	From solution, remove undissolved grumeleuse to prevent to surpass dosage
Screening	The cell type screening				The screening haemocyte (can function of use based on the cell screening of the on-hand quantity of given antibody to extract blood plasma from blood
				The particle of changing) the treatment screening that screening nature phagocyte is used for AIDS from blood (is for example sieved osteoclast at tissue or the dissimilar cell of organ, Gegenbaur's cell, osteoblast)

		The screening leucocyte sieves the knife-edge cell as leucocyte treatment of diseases (for example high WBC is leukaemia or lymphopenia for example, or low WBC) and is used for the treatment of knife-edge cell anemia disease from common red blood cell
			The cell mass screening	From individual cells, remove cell mass according to size screening island cell mass (diabetes)
	The cellular morphology screening	From healthy cell, remove infection cell from dead cell, removes living cells separate in the propagation with proliferating cells not
					From the sperm of non-activity, separate viable sperm is arranged
	Isolated cell from slicer	The osteocyte of from the blood of bone slice, purifying
				Colloid separates	According to screening colloids such as size, density, composition, S
	Screening is different from a kind of cell type	The myriospored spermatoblast of screening removes the most serious RBC in knife-edge cell anemia disease patient body from common spermatoblast

Figure 27 is a block diagram of describing exemplary double reaction device product purification and piece-rate system 2700 according to the embodiment consistent with the present invention.The double reaction device is used for the gathering of generation, recombinant protein product, virus and the viral antigen and the living cells of monoclonal antibody.The double reaction device is the most common to be applied in the production of different drug therapy acid glycerides.The double reaction device can make and allow cell to breed in the high concentration environment and work under ideal conditions.In the cell proliferation process, culture medium flows through this double reaction device.The waste product of these cells in this media set, and provide nutrition for cell.Then the medium that flows through this double reaction device is handled in order to obtain these products.In order to concentrate product, filter the medium that has been full of refuse by many different disposal processes, one of them is by the particle extraction chromatography.This processing process expends for a long time, and very expensive, and the output final products do not account for big ratio.The invention solves all these three problems.

Dual branch answer device product purifying plant 2700 can be fast, be easy to and from useless medium, extract the product of being wanted accurately.Useless medium can flow into screening plant by a passage 2701, and the particle solution that has covered suitable affine point simultaneously flows into two passages of sifter by second channel 2702 and all leads to first blender 2705, and they will mix therein.In mixed process, this product all combines with the point of particle.Blender flows through first Disengagement zone 2710 with the cushioning liquid of importing (input channel 2715) then.By using passive diffusion screening to utilize, these particles and selected product will and abandon by passage 2716 outputs by passage 2718 outputs and waste liquid.Passive diffusion sieve successfully be because bulky grain rests on a passage lighter molecule will be diffused into the another side particle solution of this passage then flow through passage 2718 simultaneously denaturing soln flow through second channel 2720.This two passage all will flow to second blender 2725, and they mix therein.When mixing them, denaturing soln will destroy the combination of product and particle.After mixing, solution will flow through output channel 2735 with another cushioning liquid by passage 2730, enter second Disengagement zone 2740.By using passive diffusion screening, particle will flow through foot passage 2745 and abandon as waste liquid.To flow through top passageway 2750 through the product of purifying and recover.This method of purification has reduced cost, has reduced the time, and has increased the output of final products.

Fig. 4 illustrates the holographic optical traps system 400 according to an embodiment consistent with the present invention, uses with device 100,200 or 300 usually.Can obtain when the 5th related application about the subsidiary details of holographic optics trapping.As holographic optics trapping device or system 400 described in Fig. 4, light is from a laser system incident, along shown in the downward direction of arrow enter activating system 400.

Phase diagram optical element 401 preferably drives attitude surface optical element (DOE), this dynamic surface also is " the PAL-SLM series of X 7665 " that the spatial light modulator (SLM) of only phase place is for example made by Japanese Hamamatsu, and is equal by " SLM512SA7 " or " SLM512SA1 " that the BoulderNonlinear Systems Nonlinear Systems of Colorado Lafayette makes.These dynamic phasings figure optical element 401 is to make the encode hologram in the medium produce light beam by computer control, and these media can change the form that produces light beam and select light beam.The phase diagram 402 that left part produces under Fig. 4 produces the trap 403 that is full of the diameter 1 μ m silicon ball 404 that is suspended in the water 405 shown in the lower right section.Like this, system 400 is controlled by the dynamic hologram shown in the bottom, left side.

Laser beam scioptics group 406,407 is transmitted to dichronic mirror 408.Beam splitter 408 is made of dichronic mirror, photon wave band crack mirror, omnidirectional's mirror or other analogous instrument.Beam splitter 408 optionally reflects and is used in light wavelength that forms optical trap 408 and the wavelength of propagating other.The then zone by the encode phase diagram from the part of the light of the regional reflex of beam splitter 408.From this part light of beam splitter 408 reflections and then by being distributed in the encoding phase figure optical element location in the plane that cooperates with the planar backside aperture that focuses on (object lens) set of lenses 409 basically.

Before the present invention the 5th related application, to learn in the trap (also claiming laser or optics pincers) at the single beam of having conceived, high power bore lens are necessary for acceptable optical trap.For the optics trapping, a basis of this imagination is these particles of gradient trapping of people's electric field of utilizing incident light.For big trapping power being arranged, having recognized that bigger electric-force gradient (or number density of light) must be arranged.The mode that people finish these usually is to pass through light field by the high-NA microlens.

Concern to sample observation and trapping in the big visual field is that this observation and trapping comprise the object lens that have low numerical aperture.Opposite with previous instruction, the numerical aperture lens are hanged down in the invention provides of the 5th related application, for example, and the object lens 409 among Fig. 4.The ability of observation and trapping can be all to be useful in any application in the case, people will benefit from owing to the big visual field of using the low power enlarging lens to have in application, for example settle the part of micro-manufacturing or for a large amount of object work, for example cell.

As example according to the present invention, the low numerical aperture trapping that the silicon ball 104 that is suspended in 3 micron diameters in the water 105 is not stated by set of lenses 109 and front.Employed lens 109 are the NA by Nikon (a) plane 4x and 0.10; (b) plane lOx and 0.25 NA produce.

Suitable phase diagram optical element be characterised in that they be according to the mode of the focused beam of their guiding light or other energy and transmission or reflection.The diffraction optical element transmitted light beam of transmission or other energy bundle, and the diffraction optical element of reflection reflects this bundle.

Phase diagram optical element 401 also can be according to having static state or dynamic surface is divided.The example of suitable static phase figure optics optical element comprises those and one or more fixed surface zone, for example grating (comprising diffraction grating, reflecting grating and transmission grating) hologram (comprising multicolour hologram, stencil, light shaping holographic filter, multicolour hologram), set of lenses, speculum, prism, wave plate or the like.The phase diagram optical element of static transmission is characterized as fixed surface.

Yet in certain embodiments, phase diagram optical element 401 itself is movably, allows to carry out the selection to a plurality of fixed surfaces zone by moving this phase diagram optical element 401 with respect to laser beam to select suitable zone with this.

Static phase figure optics optical element can be fixed in an axle and rotate jointly with the controlled motor (not shown).Static phase figure figure optical element has fixed surface and zone of dispersion.In other embodiment of static phase figure figure optical element (transmission or reflection), fixed table has the surface of region of variation continuously that comprises basically of homology not, or the discrete zone and the surface of region of variation combination continuously basically.

Function depend on the time the example of suitable dynamic phasing figure optical element comprise that computer generates diffraction pattern, phase shift material, liquid crystal phase shift array, micro mirror array, comprises piston mode micro mirror array, spatial light modulator, electrooptics guider, accousto optical modulator, deformable mirror, reflection MEMS array or the like.With dynamic phasing figure optical element 401, the changeable hologram of medium 405 codings that comprises phase diagram optical element 401, so that the shape phase shift, this phase shift is to make the focused beam that causes respective change in the phase outline of focused beam, and this type of changes for example diffraction or convergence.In addition, can change medium 405 to be created on the change of optical trap 403 positions.An advantage of dynamic phasing figure optical element 401 is can change medium 405 to move each optical trap 403 independently.

Dredge in the peripheral phase outline of those light beams (beamlet), from the inwardly then closeer embodiment in periphery, so that to have high-intensity optical trap be useful to forming in the periphery less than the excess filling back aperture of about 15% amplitude, this is the optical trap that contrast does not have excess filling back aperture to form.

In certain embodiments, the form of an optical trap can become point-like optical trap, optics whirlpool, bessel beam, optical bottle, optics rotary body or optics cage from its primitive form.This optical trap can move in two dimension or three-dimensional.The phase diagram optical element also has effect for the particular topology pattern that puts on laser, and for example, by gaussian model being converted into Gauss-Lagrangian pattern, with this, a branch of light becomes Gauss-Lagrangian pattern and another Shu Guang becomes gaussian model.Utilization to Gauss-Lagrangian pattern has strengthened the trapping ability greatly by reducing radiation pressure.

1. imaging system

Current instrument design uses a high resolution CCD camera as main imaging system 110.The major advantage of CCD camera (seeing Fig. 5 label 511) is its good ratio of performance to price, because this is a mature technique.Another advantage of CCD camera is its broad dynamic range and generates numeral output easily.

Image is presented on the computer screen (seeing Fig. 5 label 510), and with the reference frame that provides to be used to select the optics trap location, the possibility that the operator is exposed to unintentionally under the laser is reduced to minimum.

2. interface

A. object shows

User interface comprises a computer screen, and it shows the visual field that is obtained by the CCD camera.The user specifies the position of optical trap with mouse.The option that the delete position is also arranged.

Describe in more detail as following institute, the user can also specify the power of each optical trap to avoid damaging sample.In addition, it can also change the power of optical trap, because trapping depends on the difference of refractive index between sample and the suspension media, can expect, this will be different with sample.

B. hologram

Specify the purpose of optical trap position to provide the input that hologram calculates.Hologram is the function that its Fourier transform produces required optical trap array in itself.Yet this function is phase object (that is, a kind of do not absorb the object that any energy just can change wave surface) under the situation of liquid crystal display.

C. select the method for a complete set of optical trap

Usually people wish with optical trap along the specific direction mobile object.This can generate straight line (by towing) by mouse and finish.Computer program explain this straight line for along its sequential deployment a series of lean on enough near optical trap, small step moving target and be unlikely to lose locking like this to target.

The present invention also comprises the ability that optical trap changes height that changes.If laser beam is parallel to the optical axis of object lens 409, optical trap will be in the place formation contour with the focal plane of lens 409 so.Finish the change of optical trap height by adjusting hologram, make the light beam that forms optical trap when entering microscopical object lens 409, converge (or dispersing) a little like this.The height of adjusting optical trap might use lens, adjusts but have only the holographic optics trapping to allow height to each independent optical trap to be independent of other optical trap.This is to go to adjust the phase place that is produced by liquid crystal by computer program to modulate and finish.

3. sample fixer

A. summary

Sample room 700 of the present invention (seeing Fig. 7 A and 7B) is cheap and disposable.Describe although sample room 700 of the present invention is existing below, another object of the present invention be create a kind of can be with using the different flexible designs that change.Except that sample room 700, the present invention can utilize other various separation phases, and for example equipment 100,200 or 300, or with reference to Figure 13-24 and 26-27 other following unitary part.

Sample room 700 is positioned on the surface of slide 701.There is series of passages 703 sample room 700 to introduce sample or object.Passage 703 is linked to each other with the collection reservoir with supply by tubule 704 (commercial on sale).Sample or object will be suspended in the fluid media (medium) and through passage 703 and introduce the working region.Sample room 700 is covered with by slip lid 705.

B. the manufacturing of sample room

In the embodiment that the present invention includes, use a kind of polyethylene (dimethyl siloxane) (PDMS) resin make sample room 700.This process relates to the shape of using standard C AD/CAM method to generate required passage 703 on computers, and this shape is changed into a photomask with habitual photoresistance/etching technique.This photomask then generates the shape opposite with passage as negative film, is etched on the silicon wafer.The degree of depth of passage 703 is controlled by etching period.This silicon wafer is the negative replica of actual sample chamber 700.Last step comprises the sample room 700 that generates reality by cast PDMS and polymerization on wafer.This causes a PDMS casting mold to be bonded on the sheet glass 701, is covered with slip lid 705 on it.Adhesion between glass and the PDMS is carved by the oxygen candle that acts on exposed surface and is reached.

Many additional steps are to guaranteeing that steady quality is necessary.For example, fusing/sclerosis of PDMS all keeps in a vacuum to prevent to produce bubble.Silicon wafer clings wafer through silanization to prevent PDMS.There is plurality of step to relate to and cleans duplicate and keep suitable environment control.These have represented standard technology.

The little syringe needle 706 of passage 703 usefulness links to each other with micropore conduit 704, and syringe needle is fixing with glue 714, inserts among the PDMS to cast the cylindrical The Small Well 707 that links to each other with each passage 703.Sample solution is introduced passage 703 with micropump 708.

Fig. 7 B has shown a kind of illustration of introducing the exemplary configurations of sample through syringe pump 708 at 710 places.Medium is introduced 711, and refuse is collected at 71 places, and is needed in 713 collections.

The piece image that Fig. 8 has represented the scanning electron microscopy of illustration among Fig. 7 B is as from the actual generation of said process.About 50 microns wide of passage, 50 microns dark.Fig. 9 has represented the piece image of the scanning electron microscopy of working region, will carry out at this operation of research sample.Illustration clearly display channel 703 is smooth clean.Although passage 703 is rectangle on cross section, it also can be designed to other shape.Passage 703 is designed to allow sample to flow into its shape by " working region " of user according to experiment needs custom design.

C. holographic optical traps

Place a plurality of trapping points with order and be that the scanned optical traps of timesharing is different, holographic optical traps constantly illuminates each optical trap.For scanned optical traps, reach the trapping power the same with the optical trap of continuous illumination, it must provide same at least time equal light intensity.This means that scanned optical traps must have higher peak strength, determine with the proportional factor of the quantity in trapping zone at least by one.Higher peak strength has increased the chance of the photic damage of material production of institute's trapping.This damage can be produced by at least three kinds of mechanism: (1) single photon absorbs and causes hot-spot, and (2) single photon absorbs and causes photochemical transformation and the absorption of (3) multi-photon to cause photochemical transformation.Incident (1) and (2) can be by selecting light wavelength, and this wavelength is that the material and the faint wavelength of the absorption of fluid media on every side of institute's trapping alleviates.Incident (3) is a more general problem, comes work partly to alleviate with longer wavelength light.Therefore, holographic optical traps can more efficiently more gently be operated meticulous material, it is by distributing the less power that continues in the each point of an object, rather than applies with all strength or apply more high strength on one point so that cause the latent lesion object in a period of time.

Comprise in the embodiments of the invention at one, usefulness standard C AD/CAM computer program design, this design is flexibly, the passage 703 of Any shape is all programmable.As long as can not interact from getting enough far away making between the passage 703, the complexity of shape is not a factor.As seeing among Fig. 7 B and Fig. 8, organizing passage 703 can obtain easily, so can use a slice to do experiment more than once.In addition, in case produce a mold, just can be used for making thousands of sample rooms, so the method is easy to become the technology of large-scale production.According to estimates, the marginal cost of single sample chamber will be low to moderate several cents during large-scale production.

4. optical system

The earlier version of holographic optical traps uses the fixing hologram that is made of multiple material.This is enough to the principle that proof use hologram generates hundreds of optical traps.But the major defect of these holograms is them is static, and makes a hologram and will use a few hours.The LCD that has produced computer drives along with the progress of hardware makes and repeatedly generates hologram in the per second and become possibility, and optical trap is utilized as dynamic device becomes actual reality.Software control allows to control the laser beam pincers to reach the automation of separation and trapping by simple performing a programme.The principles illustrated of computed hologram is as follows.

B. microscope

Optical system 410 comprises the high quality optical microscope of standard.Objective lens is a high numerical aperture lens 409, is coupled with the long-focus condenser lens.High numerical aperture lens 409 is used for trapping.Although the long-focus condenser lens may reduce the resolution ratio of image a little, this does not jeopardize trapping and provides exceptional space to hold pipeline and plug near slide.Object can move by catching them.

Comprising in the embodiments of the invention at one, is to produce 200 microwatts on optical trap with the laser power of about 2mW.The power level that obtains from the laser instrument of a 2W enough produces about 1000 optical traps.Here used green laser (523nm), but the laser of other wavelength can use also, comprise for example far infrared laser, be used for material work with near absorbing wavelength light 532nm.

Trapping depends on refractive index gradient, refractive index and the approaching higher optical trap of material require power level of medium so on every side.In addition, material is different with trap power to the tolerance of damage, and it is desirable therefore can controlling this parameter concerning the user.The user can use the power level of any specific trap of " power slide " rising that shows at graphical interfaces.

C. liquid crystal hologram figure (being also referred to as spatial light modulator or SLM)

Spatial light modulator 408 is a liquid crystal array by electric field controls in essence, and electric field is controlled by computer program again.This liquid crystal array has the characteristic of the phase place that postpones light, and retardation depends on the electric-field intensity that is applied.

Nematic crystal equipment is used to show or the application of the only phase place modulation depth (2TT or bigger) that needs are big.The nematic crystal molecule is parallel to equipment surface usually and places to give the maximum delay effect, owing to the birefringence effect of liquid crystal.When applying electric field, molecule tilt causes and is parallel to direction of an electric field.Along with the enhancing of voltage, reduce the minimizing that causes equipment to postpone significantly along special refractive index index and birefringence.

D. laser instrument

Useful laser instrument comprises solid-state laser, semiconductor pump laser device, gas laser instrument, dyeing laser instrument, alexandrite laser instrument, free electron laser, VCSEL laser instrument, semiconductor laser, titanium-sapphire laser, the YGA laser instrument that mixes, the YLF Lasers device that mixes, semiconductor pump YGA laser instrument and flashing lamp pump laser.Semiconductor pump Nd:YGA laser instrument preferred operation at 10mW between the 5W.The laser beam preferred wavelength that is used to form the research biologic material comprises infrared ray, near-infrared, red visible light, green visible light and visible blue, and wavelength is best to about 1060nm from about 400nm.

Fig. 5 is the schematic diagram that is used to sieve the holographic optics trapping system of object, and uses with equipment 100,200 or 300 according to an embodiment consistent with the present invention.In such embodiment, optics trapping system 500 (see figure 5)s (for example BioRyx system of being sold by Chicago, Illinois city Arryx company) comprise Nixon TE 2000 series microscope 501, have settled a base that forms optical trap with holographic optics trapping unit 505 within it.Nosepiece 502 is connected on the frame, directly is contained in microscope 501 by this base.For imaging, above object lens 504, provide a lighting source 503 to illuminate sample 506.Consistent with the present invention, sample 506 is one of separation phases of equipment 100,200 or 300.

In one embodiment, optical trap system 400 (seeing Fig. 4 and Fig. 5) comprises an end of first optical channel, and this passage is very approaching at optical element, and the other end of first optical channel and second optical channel intersect and connect each other and form quadrature there.Second optical channel is formed at microlens and installs in the bottom of turntable or " Nosepiece ".This Nosepiece is suitable for being installed in Nixon TE 200 series microscope.Second optical channel connects with the 3rd optical channel, its same and second optical channel quadrature.The 3rd optical channel begins to traverse the bottom of Nosepiece and parallel with object lens focusing lens 409 from the top surface of object lens dresses parallel operation.Focusing lens 409 has top and the bottom that forms back aperture.Insert in the 3rd optical channel is a spectroscope beam splitter 408 between second optical channel and focusing lens back aperture.

The other parts that are used to form optical trap in the optical trap system comprise: first minute surface, and its reflection is by the light beam that sends from phase diagram optical element 401 of first optical channel; Be installed in the first optical transition assembly 406 in first optical channel, arrange good this assembly to receive the first minute surface beam reflected; Be installed in the second optical transition assembly 407 in first optical channel, arrange good this assembly to accept to see through the light beam of the first convertible lens group; And second minute surface 408 that is positioned at first optical channel and the second optical channel intersection point place, it is mounted the light beam of the reflecting ﹠ transmitting second optical transition assembly to enter the 3rd optical channel.

In order to produce optical trap, beam of laser directly penetrates (see figure 5)s from laser instrument 507 and passes pointing instrumentation and reflect away by optical fiber connector 508 and from the dynamic surface of diffraction optical element 509.The dynamic surface diffraction of the diffracted optical element of light beam that leaves the pointing instrumentation end of optical fiber is dispersed into a plurality of Shu Guang.The number of every Shu Guang, type and direction can change and control and change by being encoded in hologram in the dynamic surface medium.This light beam and after first minute surface reflection by the first optical transition assembly, arrives second minute surface along first optical channel by the second optical transition assembly; And aim at spectroscope 509, and up to the back aperture of object lens 504, converge through object lens 504, produce the required optical gradient condition of optical trap that generates with this.Part light after spectroscope 509 divisions, for imaging, the optical data stream (see figure 4) is formed at the bottom of passing the 3rd optical channel.

The spectrum analysis of biologic material sample can be finished by an imaging source 503, and this light source is fit to spectrum analysis or polarised light backscattering, and the former helps to evaluate chemical characteristic, and the latter is fit to measure the yardstick of internal structure, for example atomic nucleus size.Use this spectroscopic analysis methods to inquire after cell in certain embodiments.Computer 501 can be used to analyze spectroscopic data, and the identification cell has X or Y chromosome, or doubtful cancer cell, precancer cell and/or non-cancer cell type, or for example discern various haemocytes.Computer program then can be applied to information the guided optical trap to comprise selected cell type.Then the cell that is comprised can be identified according to reaction, or bind the cell that is comprised with other chemical substance.

This method and system help to realize following the tracks of the motion of each optical trap and the semi-automatic or full-automatic process of content.Motion can be monitored by video frequency pick-up head 511, frequency spectrum or optical data stream, and provide a computer program to control the selection of cell and the generation of optical trap.

In another embodiment, the tracking of the motion of pair cell is based on the predetermined motion of each optical trap that produces by the phase diagram optical element is encoded.In addition, in certain embodiments, a computer program is used to preserve the record of contained each cell in each optical trap.

Optical data stream just can convert vision signal to then, and the operator uses spectroscope and/or video-frequency monitor to carry out visual observation, monitors or analysis.Optical data stream can also be handled with monitor intensity by the viewpoint detector, or other suitable device is converted into optical data stream the numerical data stream that is suitable for the computer use.

Do not use in the method for SLM (spatial light modulator) one, move through object is relayed to second, third, goes to realize moving the 4th group or the like then from first group of optical trap.For object is moved on to the another location from a position, a static phase figure optical element spins the cone rotation around one makes laser beam aim at another zone, will produce second group of optical trap on corresponding second group of precalculated position in this zone.By constructing second group of optical trap suitable near primary importance, probe can be transferred into second group of optical trap from first group of optical trap.Order can continue to transmit three groups of prepositions of probe to the from second group of precalculated position continuation, position from the four groups of predetermined optical in the 3rd group of position to the, and from the 4th group of preposition or the like, the rotation by the phase diagram optical element is to aim at the suitable zone corresponding to the position of wanting.The termination and the time interval between the generation of next at one group of optical trap have one period duration to guarantee that probe was passed to next and organizes optical trap before they are free.

From one wide to the staggered moving process of narrow near zone, the staggered of cell carries out in a similar fashion at object.Yet transmitted and moved second and subsequently position from first group of optical trap to second group as object, the interlaced arrangement of trap allows object by intensive trooping, rather than, may cause that like this object is contained in the wrong optical trap simultaneously too near near the position of two objects.

In case object or cell have the interaction with trap, spectral method just can be used to observe cell.Those have positive result cell (promptly with label reaction or the cell that combines with it) spectrum can by utilize the imaging illumination to obtain for example to be suitable for stiff spectroscopy or polarised light backscattered.Computer can analysis spectral line the target wanted with identification of data and the target wanted with segregation of guiding phase diagram optical element.Be right after finishing of test, can make the selection that abandons those cells and collecting cell by computer and/or operator.

Optical peristalsis (seeing Figure 13) is existing operation, this operates in 1301 arrangements of microstream passage and uses the parallel lines of trap 1300 so that online spacing, when first trapling was closed, allowing the particle 1302 of trapping was that a line is to be drawn into the trap on another line.Optical peristalsis can be utilized as the possibility that uses with fluorescence labels (as described in application corresponding hereinafter).This is handled by regulating regularly the elimination operation of trapling, so that be moved towards the direction particle of wanting by the arrangement appointment of trapling.By the line whether opposite side of selecting particle is opened or closed at the trap of a side, particle can be in a direction by moving forward or backward.By using a large amount of traps, thus a large amount of particles can with given consistent direction on move.Like this, the particle that attracted to trap is movable to given zone, and, if desired, collect there.This handles and also can be used in and install in 100, the 200 or 300 common multiple fluid streams that use.

Same pass through gradually minimizing between the straight trap of assigned direction spacing and/or change the trap curvature of a curve, particle can be swept into a focus form with concentrated they.Otherwise such shape will be disperseed these particles.

Spacing between trapling can be relatively large to quicken moving of particle, and perhaps relative narrower is so that its deceleration.Similar, changing the density of selecting trap or line, they act on particle thus, also can use.By collecting or diffusion liquid stream, particle can make up or separate.In addition, optical peristalsis can combine with the differential effect of viscous drag or electric field and think that good parting material generates complicated and special parameter value group.By opposite trapping and other power, two equilibrium of forces points determine that particle still is that another power moves together with trap.

In an embodiment consistent with the present invention, optical peristalsis can be carried out together with the holophotal system that circulates by the phase diagram order with the order of carrying out corresponding holographic optics trapping shape.Such shape can be that wherein each shape rotates to certain position by motor according to the element encode that is arranged on the reflective, diffractive optical on the prismatic surface.Similarly, the element of the diffraction optics of transmission can be placed on the periphery of dish and rotate to browse shape.Also can use switchable phase grating of encode and phase hologram on video disc.

Because particle is handled by linear array by the angular force of outside, fluid stream for example, in trapping power significantly greater than the place of external motivating force, the trapping particle.In the bigger place of angular force, particle liquid stream is by array, and between these limit, angular force surpasses trapping power to be used for the diversity factor that the particle difference separates, and causes particle to jump to trap along the major axes orientation of array from trap.The clean deflection of zero degree can rotate to 45 places of spending at array and can be observed, because: (1) positive and negative displacement equiprobability takes place; Perhaps (2) particle is locked [11] direction, and the jump diagonal is by this array.

By the particle deviation ratio of the bigger angle of array deflection particle deviation to a bigger angle because of bias force.Optical gradient forces put on particle along with and change and to be approximately a3, a=radius herein.The Stokes power that acts on the particle changes with " a ".Like this, the bigger particle array of traps that is subjected in various degree influences, and the less deflection of smaller particles experience.Be oriented in array near the optimal deflection angle array and calibrate density and under the drift condition, settle the largest particles, therefore, here have at bigger inflection point than granule, the particle of deflection can be collected or further utilize the additional arrays in first downstream to separate in various degree.

Some are used to separate conventional method and finish separation in the direction that applies power.Yet this technology is not continuous to sample operation in batch.

Other conventional method that is used for micro-separation has been used the filter screen of the micro-moulding of two-dimensional crystal lattice barrier or fence formation.For example, the Brownian movement of the particle that passes filter screen is proofreaied and correct in the arrangement of a uneven barrier, causes these particles along being moved by the path that this particle diffusivity determined.Yet, be not adjustable to the easy obstruction of use of micro-moulding lattice and according to granular size and type.

In Figure 10, be proof for example according to the example of particle screening of the present invention.Though illustrative example illustration lateral deflection, optical peristalsis can obtain at same system.The representative of video image has shown that the material based on light separates, in this case, then based on the granular size separate object.Liquid stream in the passage of last left part comprises the particle of 1,2.25 and 4.5 μ m, and another liquid stream enters from following left part.These overlapping circuits are represented every liquid stream of passage respectively, when system's Laser Power Devices are closed.When Laser Power Devices were opened, light entered interaction area (with overlapping green square frame indication), extracted 4.5 μ m particles and was sent to passage with the overlapping lower right section that white path indicated from top liquid stream.

6. the haemocyte screening is used

A. background

In an application consistent, use a high-resolution that utilizes optical trap technique, the cell sifter of high penetration with the present invention.For the needs of implementing this technology as the new basis of cell screening, the failure proof of traditional flow cytometer is carried out cell characteristic necessity in many screening problems high-resolution needs really.

B. use the screening of holographic optical traps

The method of the cell screening of execution high-resolution of the present invention, high-throughput has following composition: the exploitation of micro-Flow Control, the exploitation of optical trap system (the trapping assembly and the trap assembly that is used for piece-rate system that are used for the funnel system), high-resolution fluorescence measurement instrument, system's control (comprising that hologram calculates) and principle design.

First assembly is to have the liquid stream input channel of transporting the input sample to separate the back cell goes out two output channels from input channel liquid stream unit with transmitting.Second assembly is the trap (being somebody's turn to do " funnel function " is that the jet pipe formation droplet that equates flows into a traditional flow cytometer) of one group of enforcement " funnel " function.The 3rd assembly is a detection system, and last, and the 4th assembly is screening system.Figure 11 A-11B shows the relation of these four assemblies.Can use and install 100, the 200 or 300 fluids streams that use together and implement similar functions.

The necessary characteristic that the embodiment of the invention of permission suggestion is finished high-throughput is that its inner capacities is operating material under the hand-to-hand situation each other with parallel line simultaneously.For initial enforcement, have 10 running systems by each incoming line 1100 that causes 10 microns separation.This is provided with the liquid stream integral width that enters from 110 microns input reservoir.Output channel 1102,1103 equally is 110 microns wide separately, and is identical with input channel 301, and they and input channel 301 extend in parallel, shown in Figure 11 A and 11B.What introduce " output channel " 1102,1103 is cushioning liquid, that is to say that the identical flow velocity to keep in input channel 1101 injects this passage.Three of all these passages 1101,1102,1103 be used for keeping surpassing the laminar flow of interested range of flow.Also can utilize above-mentioned about installing 100,200 or 300 screening step.In screen area, specific cells is sent to one of output channel 1102,1103 from input channel 1101, and three all liquid streams are adjacent, between do not have machinery to separate.Laminar flow remains on their arbitrary materials in the liquid stream separately, unless introduce specific external force so that material is sent to another from a flow channel.

Funnel trap 1105 acts on input cell 1106 so that they all have the streamline of obviously determining, and input cell 1106 is separated with the beeline of being set by the operator each other.Passage 1101,1102, the flow velocity in 1103 is beeline 1106 equipment thus, and " renewals " speed by carrying out separation function equipment and whole cell handling rates of requirement determine.

The funnel system is by being arranged on the runner so that the low-intensity trap 1105 that shape is set up as the static hologram of pivoted configuration function is formed with one group.The funnel trap intensity and the fixed-site in downstream, only provide with the trap 1105 that keeps the separation upstream between the cell streamline be allowed to change in time intensity and position and do in order to interference condense on the cell liquid stream and allow cell single, that do not condense pass through.

The measurement of determining screening occurs in the catchment of funnel trap 1105 or occurs in the farther zone outside the funnel system.For this starter system, mensuration will be made up of the high-resolution fluoroscopic examination.In the future, can implement other the screening standard that works, such as using scatterometry, perhaps by braking technique, such as those of deflection optical of utilizing listed earlier.

The final assembly of device is an autonomous system, screening standard therein be utilized with transitional cell enter output channel 1102,1103 one of them or allow them to be deposited in the liquid stream of input channel 1101.The key parameter of this assembly is to be used to implement the visual field that dynamic trap 1105 arrays are carried out the high-NA objective 1104 that separates.The width of visual field is the same with single width of channel to be 110 microns.Length, yet, depend on flow velocity, channel depth and be used to control the renewal speed of the Optical devices of trap.

Now, an embodiment consistent with the present invention comprises being created in to drive in the optics trapping system that the spatial light modulator of phase mask is efficiently arranged.The renewal speed of these devices is at 30 hertz or higher.The estimating channel degree of depth is 10 microns, and sets spermatoblast and should move 1 micron step, and spatial light modulator upgrades for 10 times and is used to move from input channel 1101 centers the center of a cell to one of output channel 1102,1103.A updating value is 30Hz, and the execution of these 10 steps will be present in 1/3 second.Under the flow velocity of 3 mm/second, these 10 steps are implemented on 1 millimeter length towards flow direction.Therefore the object lens 1104 that are used for separation assembly have the workspace of 110 microns x1000 microns.The important improvement field of this project is the design of this set of lenses.Balance in the lens design is between visual field and the aperture multiple usually.That is, for special complicated lens group, an important in these areas performance increase will reduce appearance simultaneously with performance in yet another aspect.This is quite complicated reason with the lens of the high performance in the print production zone of for example high-resolution integrated circuit electronics just.Yet the present invention does not require the over-all properties level of these set of lenses.

7. disclosing aspect the vortex light pincers of wide field

Light pincers under wide visual field comprise having low numerical aperture microcobjective.The ability of optical trap object is axially depending on at the downward focused beam of the mode that greatest gradient is axially arranged.Here hint forms light cone the wideest used radius.The radius of cone directly is decided by the numerical aperture of object lens, and promptly high-NA means wide cone radius.This and direct conflict of wide visual field necessary condition.The axial wide visual field light pincers that make of this routine become difficult.Causing the main cause of axial light pincers difficulty is the radiation pressure of focused beam.Especially on density with the particle of surrounding medium optimum Match, polystyrene microsphere body for example, radiation pressure can go out trap with granule impact.Under low NA objective, be difficult to overcome the radiation pressure that has axially enough light pincers power.Yet holographic optical traps has the ability to form the optical mode of the radiation pressure that significantly reduces light beam.The vortex trap, for example, having in the dark, the heart changes phase place in the counteracting of the center of trap because of light.This in the dark the heart mean that most of light of propagating downwards along beam center no longer exist.These light beams contain the radiation pressure of most of light just, so their removal has alleviated the difficulty of axial trapping greatly.Other mode, promptly the annulus mode has identical advantage.

Usually be safer for manipulation object or cell (pushing away guiding, screening) by obtaining multiple light beam.As nail matrix, multiple light pincers guarantee to introduce any specified point of driving force in cell still less.This has removed heat spot and has reduced damaged risk.Any harmful two-light quantum is handled very big benefit, because absorb square in direct ratio with laser power.Only increasing by second tweezers reduces at specified point 1/4th of two amount photonic absorption.

Finally, by utilizing holographic optics trapping, even also be greatly enhanced for the manipulation of single cell only.Single cell can clamp line (tweezerslin) operation by light, this pincers line in a side along this cell of all long supports.The rotation that causes allows the observation of 360 degree to this cell.In addition, be used for the advantage that biological sample is observed, also have the ability of firm directional sample, this for example has very strong dependent scattering experiment beneficial to sample orientation for research significantly.

8. based on the cell sifter that rotates dish

Utilize laser to exist with the form of rotary laser dish, CD player or DVD player with a large amount of zones of visit rapidly.These equipment comprise the rotatablely moving of dish of the radial motion that has fabulous high-rate laser access region.For example, typical DVD player can be visited about 4,000,000,000 independently " positions " on dish in about two hours.Method and optics trapping (the seeing Figure 12) combination of this rotation dish is allowed to visit cell with similar speed, and the holographic optics trapping is with this speed of increase of 100 times or higher multiple.

Figure 12 has described a cell sifter based on the rotation dish, according to one with of the present invention second and the 5th consistent embodiment of related application.As shown in figure 12, object or cell are introduced at sample inlet 1200, and the sample that utilizes suitable sample delivery system 1201, cell to be provided for to rotate by power management distributes dish 1202.Sieve with imaging and trapping system 1203 pair cells that control is connected with analytical system 1204, these cells are concentrated in sample room 1205 and 1206.

Principle at the surface distributed cell of dish is arranged a lot.The liquid stream chamber that holds individual cells, the gel of fixed cell, the viscosity of fixed cell and wax surface, perhaps even freeze cell and become solid block, these are operable all methods.In case celluar localization is so that they keep its relative position, they can reasonably be measured.Can use the optics trapping from surface or body, to discharge that want or undesired cell.The condition that screening therein surpasses two group is to want, and each group can remove single step, and can carry out multistep.

9. utilize lyase to carry out the screening of cell and non-biological material

Such as the technology of fluorescence activated cell screening (FACS) and so on, though set up preferably, still bearing them is facts of sequential processes method.Because in the generality of biologically labeling dye, be possible based on the screening of these dyestuffs.These dyestuffs often produce the difference in the absorption of some wavelength of dyeing and undyed sample or wave-length coverage, and there is such absorption difference in the inherence of supposing to sieve group.Thereby holographic optical traps can be used to heat and operate sample is substrate from the temperature enzymolysis that improves sample.The sample of implanting can be removed and subsequently along with the increase of bulk temperature.In addition, one faster, even more operation repetitives are possible, and wherein cell is by wide, a high power light source lighting of the integral array of handling sample simultaneously.Mutually same group of methods can be applied to non-biological materials different aspect absorption spectrum, perhaps optionally is used for this.

10. based on the screening of gel

Holographic optics laser trap has been explained the big advantage to operand, can visit on three-dimensional and mobile object with these operands.Become more advanced because the biology screening is used, through less time commonly used, more sample need be sized.The three-dimensional visit of holographic optical traps means that these screening application can realize.Step-sizing operation a large amount of cells and other trouble or impossible or the interested biological sample on two-dimentional matrix can effectively be sieved.

The enforcement of the screening of such three-dimensional relies on reversible gelation and handles.The cell gel anchors on the network, so cell that want or undesired utilizes holographic optical traps to extract from gel.Melting agarose gel and provide and withdraw from passage can be provided the heat that goes out from trap.

Perhaps, cell is optionally to be killed, based on some holographic laser trap standard.Whole gel melts with living cells separated from dead cell then.Replace only killing, a more harmful thermal explosion may produce, and this blast can make that cell is cracked to be many little parts, then may be affected according to the screening of size, once more in groups or link together with some cell.

11. kill the application of biological sample

A large amount of multiple application are from optionally killing the biological sample income.Removing pathogen from blood is exactly such application.Cell screening is an Another application.The identification cell is killed one or more cell masses, and removes dead cell.Kill by the luminous energy that comes from laser self and finish, and may not need optical trap to finish this function.

In fact, cell is heated or the medium of encircling cell is heated, destroy and cell killing with laser beam.Holographic optical traps, because their versatility and three-dimensional control allow optionally, parallel in a large number cell killing.

12. example

(Arryx company, Chicago II) can be clamped from whole blood and obtain blood platelet to utilize the BioRyx200 system.The blood platelet that pincers obtain carries out under the low laser power (0.2W) of 532nm and moves on the 3-D space easily.It is preferred using slightly high power to be used to make blood platelet to transmit by automatic trap, although 0.8W is enough.Even used anti-agglomerating agent, blood platelet to still have the short chain fibrin to be attached to them.Surpass for a long time, blood platelet may inevitably combine with cover glass.Blood platelet is approximately the 2-3 micron dimensionally, and they almost under 532nm the silicon ball with the 2-3 micron obtained by pincers.When RBC is under the view coordinate system identical with blood platelet, they trend towards being repelled by unfocused light cone, even RBC is far from trap.Yet if red blood cell and laser contact, laser will puncture them and often cause their blasts, depend on the permeability and the laser power of medium.The reaction for the laser pincers of dissimilar WBC is then slightly different.Usually, WBC is repelled slightly from them.The differential responses for optical trap like this provide the basis that is used for the isolated cell type, by they differential responses for the trapping light beam.For example, blood platelet can move by trapping and by handling laser beam, and simultaneously RBC and some WBC is ostracised and other WBC by trapping and move to the centre degree.

By with above-mentioned technical combinations, it is calculated that people can be with per 20 minutes 1011 hematoblastic speed washed corpuscles compositions.The higher rate of screening can realize by further the laminar flow screening of these technology and instrument 100,200 or 300 being made up.

Above-mentioned different technologies can be used to sieve a lot of different materials.For example, for the screening sperm, sperm can be sized based on motility or activity, such as move or swim the liquid stream of selecting into one by movable sperm, perhaps by making sperm that do not move about or non-activity be deposited into waste liquid stream.Sperm also can be sized and enter a plurality of passages, and each all has different mean motions.Sperm also can be by isolated and separate in multiple different pathogens or other the undesired material from the sperm mixture.Also can utilize above-mentioned separation cleaning and/or cooling procedure.In addition, output or movable or activated sperm can be enhanced, and for example, the temperature by controlling different liquid stream and the chemical composition of liquid stream are such as by increasing derivant or protective agent.

Figure 25 has described the activity of sperm of ox or the screening of movement, utilizes various embodiments of the invention, and wherein a high movement property and active sample produce from the sample of SA and motility.Solidify that seminal fluid is thawed and in salt solution rinse and remove glycerine, test yolk, and other material so that the density that is complementary with cushioning liquid or salt solution, PEG and BSA to be provided, are fluently used by second liquid.Optionally, can add glycerine to buffer stream.Use 0.01 in the past to the flow velocity of 0.1ml/min, and that 0.025ml/min uses is the most general, in sifter, such as device 200.The sperm concentration of 500 ten thousand about cell/ml, wherein 5-60% or higher activity arranged is utilized, as input liquid stream solution.After screening, find that selected liquid stream has been enhanced 80% activity is arranged, and outcome expectancy reach have near 90-100% active and movable.

Also can utilize other different sifters configuration, and improve the result and also can laser steering be realized with using based on the screening of laminar flow by utilizing.For example, the speed with respect to inlet flow of increase buffer stream has increased the width at the buffer channel of Disengagement zone, reduce the distance (and increasing the distance that withdraws from buffer stream) that sperm must move into buffer stream, be increased in the output (as selected liquid stream) in the buffer stream.Increase the waste fluid channel flow velocity and also can improve output, make the Necrospermia near any cushion input channel introduce waste fluid channel once more.

Above-mentioned screening also can utilize gradient with the raising screening efficiency, and different liquid stream has different characteristics, for example produces the gradient, thermograde, velocity gradient, viscosity gradient and diffusion gradient.

Except that screening, also can utilize different embodiment of the present invention to change the cell concentration of particle, for example, such as the granule density that increases in the selected liquid stream, perhaps diluted concentration by input buffering solution.Diffusion coefficient also is exercisable, the object diffusivity (perhaps movement) of change in different independently liquid stream, such as passing through to change temperature, chemical concentrations, the viscosity of liquid stream, liquid current density, salinity, use surfactants etc. for example, change the physical attractiveness of liquid hydromechanics radius or object by the time.

Therefore, according to the present invention, many holographic optical traps, can these traps of independent operation, can use with device 100,200,300, with operation composition or particle, such as haemocyte and other blood constituent, flow to another liquid stream from a liquid, become the part of independent process.For example, can discern in the interested liquid stream one composition and move to liquid stream 2 by holographic optical traps, and with this component separation in another liquid stream 1.

Fig. 6 shows the flow chart of a method embodiment of the present invention, and a useful summary is provided.With initial step 600 beginnings, this method provides the stream of first liquid with multiple composition, and step 605 is such as multiple blood constituent.Second liquid stream is provided, step 610, and first liquid stream contacts so that first Disengagement zone, step 615 to be provided with second liquid stream.First composition divides and is deposited into second liquid stream different in naturely in the multiple composition, step 620, and second composition in the multiple composition remains in first liquid stream simultaneously, step 625.Step 630 is removed in second liquid flow point opposite sex ground with first composition from first liquid stream with second composition.When separation (perhaps step) existence that does not have to add, step 635, the method can stop returning step 680.

When other separation existence, step 635, the method proceeds to step 640, and the 3rd liquid stream is provided.First liquid stream contacts so that second Disengagement zone, step 645 to be provided with the 3rd liquid stream.Handling when holography will be in second line bonus when utilizing, step 650, and the method proceeds to step 655, and many holographic trap generations, typically utilizes optical wavelength.Utilize holographic trap, second composition in the multiple composition divides and moves into the 3rd liquid stream, step 660 different in naturely.When holography operation is not used in second auxiliary the separation, step 650, the method proceeds to step 665, and second composition of multiple composition divides and is deposited into the 3rd liquid stream different in naturely.After step 660 or 665, the 3rd composition in the multiple composition remains in first liquid stream simultaneously, step 670.The 3rd liquid stream with second composition then divides opposite sex ground to remove from having the ternary first liquid diffluence, and step 675 and the method can stop, and return step 680.Though in Fig. 6, do not illustrate, should be understood the method and can be continued on for other separating step, such as third and fourth, five, or the like.

Other embodiments of the invention are described in Figure 14-24.Figure 14 and 15 has described the different screening systems 1400 and 1500 of the different screening step of operative installations 100,200 or 300, comprises the reservoir and the moving pump that are used for fluid stream.Figure 16 is a side view, and Figure 17 is the plane of high-aspect-ratio plane sifter.The sifter of this high-aspect-ratio can be used in provides big relatively laminar flow liquid stream parting surface between different liquid stream, and a bigger contact area that is used for the component separation between liquid stream is provided.

Figure 18 one has the perspective view of the three-dimensional screening plant of multiple plane sifter (having illustrated, such as sifter 1600).Figure 19 is multichannel sifter 1700 planes.Figure 20 is the plane with sifter 1800 of narrow waste liquid flow region 1801.Figure 21 is the plane that is used for the sifter 1900 of the different flow velocity of utilizing of different passages.Figure 22 is the plane with sifter 2000 of multiple selector channel 2001.Figure 23 is the plane with sifter 2100 of compression screen area 2105.

Figure 24 A, Figure 24 B are respectively the side view and the planes of multilayer laminar flow sifter 2200, and wherein input channel is in layer 1 (2210), and layer 2 provides multiple screening step (2200), and layer 3 provides output channel (2230).These different screening steps can connect according to countless connection in series-parallel relations.Many other the mutation of the rapid sifter of this multistep will be easily apparent for those of ordinary skills.

Also can utilize the shape and the structure of other passage.For example, can utilize channel design crooked or " snakelike " so that long interaction zone to be provided within less zone or volume.In the Disengagement zone, also can utilize the flow-control bar, to regulate the fluid stream in the passage.For example, montant provides the obstacle of fluid flow, has reduced channel size and Reynolds number effectively, has also improved laminar flow.

Again in general, and for instance, first composition in the multiple composition can be numerous red blood cells and numerous leucocyte, and second composition is numerous blood platelet.In a second embodiment, numerous leucocytes can separate from numerous red blood cells on holographic formula ground, for example holographic (optics) trapping of the technology of utilization.Also can utilize holographic trapping to remove multiple pollutant from first liquid stream, perhaps holographic separating bio is learned fragment from first liquid stream holographicly.The first liquid stream can comprise basically from donor next whole blood and anticoagulant in different embodiment, and the second liquid stream can comprise basically from the next blood plasma of donor.Different sedimentations banded or isopycnic.Different liquid stream is essentially no turbulent flow, but and also laminar flow basically.

First and second Disengagement zone respectively have one predetermined fully with flow to parallel length and fully perpendicular to the predetermined degree of depth that flows to, the predetermined length and the predetermined degree of depth are definite according to second flow velocity of first flow velocity of second rate of settling of first rate of settling of first composition, second composition, first liquid stream and second liquid stream.First liquid stream and second liquid stream also can have essentially identical flow velocity.Optionally, first liquid stream also can have first flow velocity, and second liquid stream also has second flow velocity, and wherein second flow velocity, first flow velocity that compares is big.

Again in general, the present invention further comprises and is used for the device that separation fluid mixtures becomes single component, inactive composition, comprise (1) have first import (120 or 315) that is used for first liquid stream and be used for second liquid stream second import (120 or 320) first sieve passage (110 or 325); The first screening passage further has first outlet that is used for first liquid stream (130 or Fig. 3 continuous passage) and is used for second of second liquid stream and exports 130 (or 320), first composition in the multiple composition of the first screening channel adaptation in allowing first liquid stream is deposited into second liquid stream with second liquid stream of formation enrichment and first liquid stream of dilution, and simultaneously in first liquid stream, second composition that keeps multiple composition, (2) have first optics inlet and export (350) with first optics with first outlet (continuous passage of Fig. 3) that is used for first liquid stream is connected, further have second optics inlet (335) second optically transparent screening passages that are used for the 3rd liquid stream and second optics outlet that is used for the 3rd liquid stream (345), and (3) and the channel attached holographic optical traps of second optically transparent screening system (400,500), this holographic optical traps system is suitable for producing holographic optical traps to select and to enter the 3rd liquid stream from first liquid moving second composition that drifts.

The another kind of device or the system that are used for the multiple composition of separation of the fluid comprise: optical clear screening path 10 0,200 or 300, have first import that is used for first liquid stream and second import that is used for second liquid stream, optically transparent screening passage further has first outlet that is used for first liquid stream and second outlet that is used for second liquid stream; With with the channel attached holographic optical traps of optically transparent screening system, holographic optical traps system 500 is adapted to produce holographic optical trap to select and to move first composition of the multiple composition in first liquid stream in first liquid stream, enter second liquid stream and form second liquid stream of enrichment and first liquid stream of dilution, and second composition in the multiple composition remains in first liquid stream simultaneously.

At last, another kind is provided for separating the method embodiment of various kinds of cell, and comprising provides the stream of the liquid with various kinds of cell; Second liquid stream is provided; First liquid stream is contacted, so that first Disengagement zone to be provided with second liquid stream; And divide first cell deposition that makes in the various kinds of cell to go into second liquid stream different in naturely, and flow second cell that keeps simultaneously in the various kinds of cell at first liquid.The method comprises also that usually the removal from first liquid stream with second cell of branch opposite sex ground has second liquid stream of first cell.The method also can be provided for providing the 3rd liquid stream; First liquid stream is contacted so that second Disengagement zone to be provided with the 3rd liquid stream; And divide second cell deposition that makes in the various kinds of cell to go into the 3rd liquid stream different in naturely, and in first liquid stream, keep the 3rd cell in the various kinds of cell simultaneously.In addition, can holographicly from first liquid stream, separate numerous second cells, and holographicly from first liquid stream, remove multiple pollutant or biology fragment.

Screening separates to produce different blood to blood constituent though above-mentioned discussion concentrates on, device of the present invention, method and system can extend to the particulate of other type, biology or cellulated material, these materials do not move about, and can or can carry out optical manipulation in deposition or emulsification in the fluid stream.For example, methodology of the present invention can be used in separates spermatoblast that do not move about or non-activity from living cells, by allowing not being deposited into second liquid stream from the first liquid stream of swarm cell.The screening of other cell separation also can be carried out, for example detached island cell or other from other the pancreatic cell of type, the island cell mass of separation different size, by the liquid flow point from optics light pincers (trapping) in both or one of them.The virus that the different rates of settling is arranged, protein and other big molecule also can separate with the present invention.Utilizing the holographic optics trapping of a plurality of separating steps can be special useful in the cell or the particle separation of these other types also.

From above, many obviously variations are effective with being modified under the spirit and scope prerequisite that does not deviate from novel concept of the present invention.Should be appreciated that plan or supposition be not about the restriction of concrete grammar described herein and device.Certainly, will make being included in claims scope by appending claims for all such modifications.

Claims (71)

1. the method for a separating blood constituents, this method comprises:

The stream of first liquid with multiple blood constituent is provided;

Second liquid stream is provided;

Contact with first liquid stream with second liquid stream, so that first Disengagement zone to be provided; And

Divide opposite sex ground that second liquid stream is gone in the first haemocyte composition sedimentation of multiple blood constituent, the second haemocyte composition with multiple blood constituent remains in first liquid stream simultaneously.

2. the method for claim 1 further comprises:

Divide opposite sex ground from first liquid stream, to remove second liquid stream with first haemocyte composition with second haemocyte composition.

3. the method for claim 1, wherein the first haemocyte composition is numerous red blood cells and numerous leucocytes.

4. method as claimed in claim 3 further comprises:

From numerous red blood cells, isolate numerous leucocytes in holographic mode.

5. the method for claim 1, wherein the second haemocyte composition is numerous blood platelet.

6. the method for claim 1 further comprises:

The 3rd liquid stream is provided;

Contact with first liquid stream with the 3rd liquid stream, so that second Disengagement zone to be provided; And

Divide opposite sex ground that the 3rd liquid stream is gone in the second haemocyte composition sedimentation of multiple blood constituent, the 3rd blood constituent with multiple blood constituent remains in first liquid stream simultaneously.

7. method as claimed in claim 6 further comprises:

With the holographic mode trapping second haemocyte composition.

8. method as claimed in claim 6, wherein, the second haemocyte composition is numerous blood platelet, and the 3rd blood constituent is a blood plasma.

9. method as claimed in claim 6 further comprises:

Recycle first liquid stream to form second liquid stream.

10. the method for claim 1 further comprises:

From first liquid stream, isolate the second numerous haemocyte compositions in holographic mode.

11. the method for claim 1 further comprises:

From first liquid stream, remove numerous pollutant or biology fragment in holographic mode.

12. the method for claim 1, wherein.The first liquid stream consists essentially of whole blood and the anticoagulant from donor, and the second liquid stream consists essentially of the blood plasma from donor.

13. the method for claim 1, wherein described precipitation step is the speed band shape.

14. the method for claim 1, wherein described precipitation step is isopycnic.

15. the method for claim 1, wherein first liquid stream and the second liquid stream are essentially no turbulent flows.

16. the method for claim 1, wherein first liquid stream and the second liquid stream are essentially laminar flow.

17. the method for claim 1, wherein, described Disengagement zone has: the predetermined length that is arranged essentially parallel to first and second liquid stream flow direction, be substantially perpendicular to the desired depth of first and second liquid stream flow direction, described predetermined length and desired depth depend on first rate of settling of the first haemocyte composition, second rate of settling of the second haemocyte composition, first flow velocity of first liquid stream and second flow velocity of second liquid stream.

18. the method for claim 1, wherein first liquid stream and the second liquid stream have substantially the same flow velocity.

19. the method for claim 1, wherein first liquid stream has first flow velocity, the second liquid stream has second flow velocity, and second flow velocity is relatively big than first flow velocity.

20. the method for claim 1, wherein first liquid stream is relatively little at second volume of described Disengagement zone than second liquid stream at first volume of described Disengagement zone.

21. device of implementing the method for claim 1.

22. one kind is separated into the method for constituent with fluid mixture, this method comprises:

First liquid stream that is essentially laminar flow with fluid mixture is provided, and described fluid mixture has multiple composition, and described multiple composition has the corresponding multiple rate of settling;

Second liquid that is essentially laminar flow stream is provided;

Contact with first liquid stream with second liquid stream, to provide first Disengagement zone, first liquid stream and second liquid stream have essentially no turbulent flow in the Disengagement zone interface;

Second liquid stream is gone into second liquid stream of formation enrichment and first liquid stream of dilution with the first composition sedimentation of multiple composition in branch opposite sex ground from first liquid stream, second composition with multiple composition remains in first liquid stream simultaneously, described first composition has first rate of settling of the multiple rate of settling and second composition has second rate of settling of the multiple rate of settling, and wherein first rate of settling is relatively big than second rate of settling;

Divide opposite sex ground that the first liquid diffluence from dilution of second liquid stream of enrichment is removed;

And

In first liquid stream of dilution, handle second composition in holographic mode.

23. method as claimed in claim 22, wherein, described fluid mixture is a whole blood, and described first composition is numerous red blood cell and leucocyte, and described second composition is numerous blood platelet.

24. method as claimed in claim 22, wherein, described fluid mixture is a sperm, and described first composition is the spermatoblast of numerous that do not move about or non-activities, and described second composition is numerous spermatoblasts that move about or activated.

25. method as claimed in claim 22, wherein, described optics maneuvering sequence further comprises:

With holographic mode trapping second composition, remove second composition with the first liquid diffluence from dilution.

26. method as claimed in claim 22 further comprises:

From first liquid stream, remove numerous pollutant or biology fragment in holographic mode.

27. method as claimed in claim 22 further comprises:

The 3rd liquid stream is provided;

Contact so that second Disengagement zone to be provided with first liquid stream of dilution with the 3rd liquid stream; And

With holographic mode trapping second composition, and with second composition from first liquid flow movement to the, three liquid of dilution stream, the 3rd composition with multiple composition remains in first liquid stream of dilution simultaneously.

28. the method as claim 27 is stated further comprises:

First liquid stream of recirculation dilution is to form second liquid stream.

29. device of implementing method as claimed in claim 22.

30. one kind is separated into the method for constituent with fluid mixture, this method comprises:

It is first liquid stream of laminar flow that basic H with fluid mixture is provided, and described fluid mixture has multiple composition, and described multiple composition has the corresponding multiple rate of settling;

Second liquid that is essentially laminar flow stream is provided;

The 3rd liquid that is essentially laminar flow stream is provided;

Contact with first liquid stream with the 3rd liquid stream with second liquid stream, so that first Disengagement zone to be provided, first liquid stream and second liquid stream have essentially no turbulent flow in described Disengagement zone first interface, and the second contact surface that first liquid flows and the 3rd liquid stream has essentially no turbulent flow in described Disengagement zone;

Branch opposite sex ground moves to first composition of multiple composition in second liquid stream from first liquid stream, with second liquid stream of formation enrichment and first liquid stream of dilution;

Branch opposite sex ground moves to second composition of multiple composition in the 3rd liquid stream from first liquid stream, with the 3rd liquid stream of formation enrichment and first liquid stream of more dilution;

Divide opposite sex ground from first liquid stream of dilution, to remove second liquid stream of enrichment; And

Divide opposite sex ground from first liquid stream of dilution, to remove the 3rd liquid stream of enrichment.

31. method as claimed in claim 30, wherein, first liquid stream, second liquid stream and the 3rd liquid flow each and all have different density, to form the gradient density gradient in first Disengagement zone.

32. method as claimed in claim 30 wherein, is removed first composition and second composition by diffusion method from the first liquid diffluence.

33. method as claimed in claim 30 wherein, is removed first composition and second composition by isodensity or the banded sedimentation of speed from the first liquid diffluence.

34. method as claimed in claim 30 wherein, is removed second composition by holographic optics trapping operation from the first liquid diffluence.

35. method as claimed in claim 30 wherein, is removed first composition and second composition by electromagnetic field from the first liquid diffluence.

36. method as claimed in claim 30, wherein, described fluid mixture is a whole blood, and described first composition is numerous red blood cell and leucocyte, and described second composition is numerous blood platelet.

37. method as claimed in claim 30, wherein, described fluid mixture is a sperm, and first composition is the spermatoblast of numerous that do not move about or non-activities, and second composition is numerous spermatoblasts that move about or activated.

38. method as claimed in claim 30, wherein, the step of dividing opposite sex ground to remove second composition from first liquid stream further comprises:

With holographic mode trapping second composition, remove second composition with the first liquid diffluence from dilution.

39. one kind is separated into the device of constituent with fluid mixture, this device comprises:

Be used to provide the parts of first liquid stream that is essentially laminar flow with fluid mixture, described fluid mixture has multiple composition, and described multiple composition has the corresponding multiple rate of settling;

Be used to provide the parts of second liquid stream that is essentially laminar flow;

Be used to provide the parts of the 3rd liquid stream that is essentially laminar flow;

Be used for second liquid stream is contacted so that the parts of first Disengagement zone to be provided with first liquid stream with the 3rd liquid stream, first liquid stream and second liquid stream have essentially no turbulent flow in the Disengagement zone first interface, and the second contact surface that first liquid flows and the 3rd liquid stream has essentially no turbulent flow in the Disengagement zone;

Be used for branch opposite sex ground and first composition of multiple composition moved to second liquid stream parts that first liquid with second liquid stream that forms enrichment and dilution flows from first liquid stream;

Be used for branch opposite sex ground and first composition of multiple composition moved to the 3rd liquid stream parts with first liquid stream of the 3rd liquid stream that forms enrichment and more dilution from first liquid stream;

Be used for branch opposite sex ground and remove the parts of second liquid stream of enrichment from first liquid stream of dilution; And

Be used for branch opposite sex ground and remove the parts of the 3rd liquid stream of enrichment from first liquid stream of dilution.

40. device as claimed in claim 39, wherein, first liquid stream, second liquid stream and the 3rd liquid flow each and all have different density, so that the gradient density gradient to be provided in first Disengagement zone.

41. device as claimed in claim 39 wherein, is used for removing from the first liquid diffluence parts employing diffusion method of first composition and second composition.

42. device as claimed in claim 39 wherein, is used for removing the parts of first composition and second composition by isodensity or the banded sedimentation enforcement of speed from the first liquid diffluence.

43. device as claimed in claim 39 wherein, is used for removing from the first liquid diffluence parts employing holographic optical traps of second composition.

44. device as claimed in claim 39, wherein, described fluid mixture is a whole blood, and first composition is numerous red blood cell and leucocyte, and second composition is numerous blood platelet.

45. device as claimed in claim 39, wherein, described fluid mixture is a sperm, and first composition is the spermatoblast of numerous that do not move about or non-activities, and second composition is numerous spermatoblasts that move about or activated.

46. the method for a separating blood constituents, this method comprises:

First liquid stream that is essentially laminar flow with multiple blood constituent is provided;

Second liquid that is essentially laminar flow stream is provided;

Contact with first liquid stream with second liquid stream, so that first Disengagement zone to be provided, the size of first Disengagement zone pre-determines based on the multiple rate of settling of multiple blood constituent;

Divide numerous red blood cells and the numerous leucocyte of opposite sex ground to go into second liquid stream, in first liquid stream, keep the numerous blood platelets and the blood plasma of multiple blood constituent simultaneously with the banded sedimentation of speed with multiple blood constituent;

Divide opposite sex ground from first liquid stream, to remove second liquid stream with first haemocyte composition with second haemocyte composition;

The 3rd liquid stream is provided;

Contact so that second Disengagement zone to be provided with first liquid stream with the 3rd liquid stream;

Divide different in naturely the 3rd liquid stream is gone in the banded sedimentation of numerous blood platelet speed, in first liquid stream, keep blood plasma simultaneously; And

Recycle first liquid stream to form second liquid stream.

47. a device that separates multiple composition in fluid, this device comprises:

Have first import that is used for first liquid stream and be used for the optical clear screening passage of second import of second liquid stream, this optical clear screening passage further has first outlet that is used for first liquid stream and is used for second of second liquid stream and exports; And

Sieve channel attached holographic optical traps system with this optical clear, this holographic optical traps system is suitable for producing holographic optical traps, to select and in first liquid stream, to move first composition of multiple composition in first liquid stream, make it to enter second liquid stream, with second liquid stream of formation enrichment and first liquid stream of dilution, simultaneously second composition in the multiple composition is remained in first liquid stream.

48. one kind is separated into the device of the composition that do not move about of composition with fluid mixture, this device comprises:

Have first import that is used for first liquid stream and be used for second liquid stream second import first sieve passage; This first screening passage further has first outlet that is used for first liquid stream and second outlet that is used for second liquid stream, the described first screening passage is suitable for allowing in first liquid stream first composition of multiple composition in first liquid stream to be deposited into second liquid stream, with second liquid stream of formation enrichment and first liquid stream of dilution, second composition with multiple composition remains in first liquid stream simultaneously;

The second optically transparent screening passage, it has first optics inlet and the outlet of first optics that is connected with first outlet that is used for first liquid stream, and this second optically transparent screening passage further has second optics inlet that is used for the 3rd liquid stream and second optics outlet that is used for the 3rd liquid stream; And

With the channel attached holographic optical traps of this second optically transparent screening system, this holographic optical traps system is suitable for producing holographic optical traps, to select and to enter the 3rd liquid stream from first liquid moving second composition that drifts.

49. a method of separating various kinds of cell, this method comprises:

The stream of first liquid with various kinds of cell is provided;

Second liquid stream is provided;

Contact so that first Disengagement zone to be provided with first liquid stream with second liquid stream; And

Divide opposite sex ground that first cell of various kinds of cell is moved in second liquid stream, second cell with various kinds of cell remains in first liquid stream simultaneously.

50. method as claimed in claim 49 further comprises:

Divide opposite sex ground from first liquid stream, to remove second liquid stream with first cell with second cell.

51. method as claimed in claim 49 further comprises:

The 3rd liquid stream is provided;

Contact so that second Disengagement zone to be provided with first liquid stream with the 3rd liquid stream; And

Divide opposite sex ground that second cell of various kinds of cell is moved into the 3rd liquid stream, the 3rd cell with various kinds of cell remains in first liquid stream simultaneously.

52. method as claimed in claim 49 further comprises:

With holographic mode ground from the first liquid flow point from the outstanding second many cell.

53. method as claimed in claim 49 further comprises:

From first liquid stream, remove multiple pollutant or biology fragment in holographic mode.

54. method as claimed in claim 49, wherein, each of first liquid stream and second liquid stream all has different densities, so that the gradient density gradient to be provided in first Disengagement zone.

55. method as claimed in claim 49 wherein, is removed first cell by isodensity or the banded sedimentation of speed from the first liquid diffluence.

56. method as claimed in claim 55, wherein, described first cell is non-activity or the spermatoblast that do not move about.

57. method as claimed in claim 49 wherein, removes first cell by own activity from first liquid stream.

58. method as claimed in claim 57, wherein, described first cell is a spermatoblast that move about or activated.

59. a device that is used to separate various kinds of cell, this device comprises:

Be used to provide the parts of first liquid stream with various kinds of cell;

Be used to provide the parts of second liquid stream;

Be used for second liquid stream is contacted so that the parts of first Disengagement zone to be provided with first liquid stream; And

Being used for branch opposite sex ground moves to first cell of various kinds of cell second liquid stream, second cell of various kinds of cell is remained on parts in first liquid stream simultaneously.

60. device as claimed in claim 59 further comprises:

Be used for branch opposite sex ground and remove the parts of second liquid stream from first liquid stream with first cell with second cell.

61. device as claimed in claim 59 further comprises:

Be used to provide the parts of the 3rd liquid stream;

Be used for the 3rd liquid stream is contacted so that the parts of second Disengagement zone to be provided with first liquid stream; And

Being used for branch opposite sex ground moves into second cell of various kinds of cell the 3rd liquid stream, the 3rd cell of various kinds of cell is remained on parts in first liquid stream simultaneously.

62. device as claimed in claim 59 further comprises:

Be used in holographic mode from the parts of the first liquid flow point from numerous second cells.

63. device as claimed in claim 59, wherein, each of first liquid stream and second liquid stream all has different densities, so that the gradient density gradient to be provided in first Disengagement zone.

64. device as claimed in claim 59, wherein, the parts that are used for removing from the first liquid diffluence first cell are by isodensity or the banded sedimentation of speed.

65. as the described device of claim 64, wherein, first cell is non-activity or the spermatoblast that do not move about.

66. device as claimed in claim 59, wherein, being used for first cell is by own activity from first liquid parts that remove that drift.

67. as the described device of claim 66, wherein, first cell is a spermatoblast that move about or activated.

68. the multiple component separation from fluid goes out a kind of device of composition, this device has the length-width ratio of length to width, and this device comprises:

Have first import that is used for first liquid stream and be used for second liquid stream second import first sieve passage; Each of first and second liquid stream has the liquid flow path direction along device length, this first screening passage further has first outlet that is used for first liquid stream and second outlet that is used for second liquid stream, this first screening passage is suitable for allowing in first liquid stream first composition of multiple composition in first liquid stream to move into second liquid stream alternatively, with second liquid stream of formation enrichment and first liquid stream of dilution, second composition of the multiple composition of maintenance in first liquid stream simultaneously; And

Length to the length-width ratio of width less than about 2: 1.

69. as the described device of claim 68, wherein, length to the length-width ratio of width from about 2: 1 to 1: 2.

70., further comprise as the described device of claim 68:

The second optically transparent screening passage, it has first optics inlet and the outlet of first optics that is connected with first outlet that is used for first liquid stream, and this second optically transparent screening passage further has second optics inlet that is used for the 3rd liquid stream and second optics outlet that is used for the 3rd liquid stream; And

With the channel attached holographic optical traps of this second optically transparent screening system, this holographic optical traps system is suitable for producing holographic optical traps, to select and to enter the 3rd liquid stream from first liquid moving second composition that drifts.

71. one kind is separated the system of multiple composition from fluid mixture, this system comprises:

Flow plate, it has a plurality of imports that are used for first liquid stream and second liquid stream and has a plurality of outlets that are used for first liquid stream and second liquid stream, this flow plate further has at least one Disengagement zone, and in this Disengagement zone, first liquid stream and second liquid stream are laminar flow basically;

A plurality of pumps are to control the flow velocity of first liquid stream and second liquid stream;

Temperature controller, it controls the temperature of first liquid stream and second liquid; And

Computer, it is suitable for providing the user to import, and separates from fluid mixture in the Disengagement zone to control multiple composition.

Images