JP2004532991A - Dynamic 3D array that can be arranged - Google Patents

Abstract

The present invention broadly relates to an array of arrayable probes for analysis of a target in a fluid.
The probes are contained within an optical trap that allows for selective alteration and rearrangement of the quantity or quality of the probes in the array. Furthermore, arrays are dynamic in that, once aligned, the optical trap allows independent rearrangement of a given optical trap and the probes contained therein.

Description

【Technical field】
[0001]
Throughout this application, various references are referenced. The entire disclosures of these documents are incorporated herein by reference in the present application to describe the state of the art relating to the present invention.
[0002]
The present invention relates broadly to arrays of probes. More particularly, the present invention relates to systems and methods that use multiple light traps to form an array of arrayable and dynamic probes, with or without substrate binding.
[Background Art]
[0003]
Until now, arrays of potentially reactive probes have been used in analysis and other chemical and biological tests and experiments. For example, arrays are often used in the fields of genetics, biochemistry and biology to analyze samples (targets) of biological or chemical substances. In many cases, analytical samples are only available in relatively small quantities. Due to the small amount of material available, microarrays have been developed that are useful for providing a relatively high density of probes in a small array for analyzing targets in small samples.
[0004]
Microarrays used in the analysis of biological materials are often referred to as biochips. Two main applications of biochips are extraction of sequence information on specific nucleic acids (whole genome of an organism, a single gene or a part of a single gene) (Patent Document 1), and evaluation of expression (Non-Patent Document 1, Non-Patent Document 2, and Non-Patent Document 3).
[0005]
Conventional microarrays are attached to the surface of a planar solid support (substrate) and consist of oligonucleotide probes having either a one-dimensional or two-dimensional configuration. Different types of oligonucleotides attach to the substrate at predetermined locations. Thus, once the microarray is formed, the location of the probes, and thus all targets that respond to the probes, is always known. Attachment of the probe is achieved either by direct synthesis of the oligonucleotide onto the substrate by a process known as in situ photolithography synthesis (US Pat. You.
[0006]
One disadvantage of such microarrays is that the one-dimensional or two-dimensional configuration limits the surface area on which the probes can be mounted and sets a limit on the density of probes for analyzing a target. In the case of DNA hybridization between a target (DNA or DNA fragment) and a probe (immobilized oligonucleotide), the rate of hybridization is controlled by the rate at which the target can contact the probe. Thus, the higher the probe density, the higher the rate of hybridization.
[0007]
A second disadvantage of such microarrays arises from their fabrication method. Once the microarray is fabricated, the type and amount of probe will be determined.
[0008]
Another approach for analyzing targets in small samples is to attach the probe to the surface of a small bead-like substrate (US Pat. Each bead with a different probe is identified by an individual label, which allows the identification of each probe and bound target by identifying which beads have which label after analysis (see US Pat. reference).
[0009]
The identity of the bead and the probe is maintained by physically moving the bead with the probe attached to the guide, capillary, groove or hole in the sheet and subsequently cleaning the bead with the target. The non-flat nature of the beads provides a larger area for the target to respond when compared to microarray probes, while determining the identity of which beads supported which probes to determine identity. Beads must be kept in a predetermined order during analysis to keep records, or bead probes must be retrieved and each bead probe inspected after analysis
[0010]
A further disadvantage of both microarray and bead analysis is that the probe must be in physical contact with the substrate. In some instances, this contact alters the probe inside and itself, or affects the process in which the probe is used during analysis. In other instances, during or after the initial analysis, a desired change in the quantity or quality of the probe if the identity of the probe is known throughout the analysis and the sequence composition can be easily changed. May have gained useful information. However, such changes are not possible in microarray or bead analysis.
[0011]
In an unrelated technique, a technique for optically trapping particles with a plurality of simultaneously generated optical tweezers is known (see generally US Pat. Optical tweezers use gradient forces on light rays to trap particles based on the dielectric constant of the particles. In order to minimize energy, where the electric field is highest, particles having a higher dielectric constant than the surrounding media migrate to the area of the optical tweezers.
[0012]
Other types of traps that can be used to optically trap particles include, but are not limited to, optical vortex, optical bottle, optical rotator, and optical cage. Optical vortexing is a gradient surrounding a region of zero electric field that is useful for manipulating particles with a lower dielectric constant than the surrounding media, or reflective particles, or other types of particles that are repelled by optical tweezers. Generate In order to minimize energy, such particles will move to the region where the electric field is lowest, ie, the zero electric field region at the focus of a properly formed laser beam. Optical vortexing provides a region of zero electric field, such as a donut hole (toroid). The light gradient is distributed radially and has the highest electric field around the circumference of the donut. An optical vortex detains small particles in the hole of the donut. Detention is accomplished by sliding the vortex on small particles along the zero electric field equipotential lines.
[0013]
A light bottle differs from a light vortex in that it has a zero electric field only at the focal point, and at the end of the vortex a non-zero electric field surrounds the focal point in all other directions. Light bottles can be useful in trapping atoms and nanoclusters that are too small or too absorptive to be trapped with light vortices or tweezers [4].
[0014]
An optical rotator is a recently described optical tool that provides a pattern of spiral arms that trap an object [5]. This class of tools can be useful for manipulating non-spherical particles or manipulating MEM devices or nanomachines.
[0015]
The optical cage (Neil, Patent Document 7) is an enlarged version of the same optical vortex. The optical cage forms a time-averaged ring of optical tweezers to enclose particles that are too large or reflective to trap with a lower dielectric constant than the surrounding media. However, unlike the vortex, no zero electric field region is formed. Optical vortices are similar in their use to optical tweezers, but operate on the opposite principle.
[0016]
[Patent Document 1]
U.S. Pat. No. 6,025,136
[Patent Document 2]
U.S. Pat. No. 5,838,732
[Patent Document 3]
U.S. Pat. No. 5,143,854
[Patent Document 4]
WO 00/61198 pamphlet
[Patent Document 5]
International Publication No. 00/71243 pamphlet
[Patent Document 6]
US Patent No. 6055106
[Patent Document 7]
U.S. Pat. No. 5,939,716
[Non-patent document 1]
Senna M et al., "Quantitative monitoring of expression patterns with complementary DNA microarrays" (Science 270 (5235): 467-70; (Oct. 20, 1995))
[Non-patent document 2]
D. J. And Winseller E. A. "Genomics, expression and DNA arrays" (Nature 405 (6788): 827-836 (2000)).
[Non-Patent Document 3]
Ekins R. And Shu F. W. "Microarray: its origins and applications" (Trend in Biotechnology, 17: 217-18 (1999))
[Non-patent document 4]
J. Aalto and M.A. J. Paget "Generating a Beam with a Dark Focus Surrounded by Higher Intensity Regions: Light Bottle Beam" (Optical Letters, 25, 191-193, 2000)
[Non-Patent Document 5]
L. Peterson, M. P. McDonald, J.M. Aalto, W. Civet, p. E. FIG. Bryant and K.S. Drakia, "Suppressed Rotation of Optically Trapped Microscopic Particles" (Science 292, 912-914, 2001)
DISCLOSURE OF THE INVENTION
[Problems to be solved by the invention]
[0017]
There is a need for analytical methods and systems that can evaluate the interaction of a probe with a target without contacting the probe with a substrate. There is a further need for methods and systems for forming arrays of reproducible and reproducible probes, and methods for maintaining probe identity throughout an analysis, regardless of probe location. The present invention satisfies these and other needs, and provides further related advantages.
[Means for Solving the Problems]
[0018]
The present invention provides new and improved methods and systems for constructing, arranging, and using a three-dimensional array of probes.
[0019]
An optical trap is created in the container. Optical traps are created by distributing a beam, such as a laser beam, on an optical element that changes the beam by patterning its phase to create a beamlet. Subsequently, the beamlets are focused by the lens to create the necessary gradient conditions for the light trap. Subsequently, a probe of known properties is added to the container. Predetermined analytical probes are selected, each of which is then selected for inclusion in an optical trap.
[0020]
The quantity and quality of the probes forming the array can be easily rearranged by using optical traps to add, discard, or replace probes. The arrangement of the probes in the array with each other is dynamic because the spatial relationship between the probes can be varied while maintaining the identity of the probes selected to construct the array. Thus, both the array and each probe can move in three dimensions, and the probes can be positioned, moved, and repositioned in the container, either entirely or separately.
[0021]
When a probe is contained within an optical trap, it is included regardless of whether it has been repositioned in the container and regardless of the `` order '' of spatial locations in the array. The identity of the probe can be maintained by knowing the identity of the optical trap being used. Further, one optical trap can pass a probe to another optical trap, tracking the chain of restraints of the probe by the optical trap, thereby determining which probe is encompassed by the optical trap. Can be maintained.
[0022]
Other features and advantages of the invention will be set forth in part in the detailed description that follows, and in the accompanying drawings, and preferred embodiments of the invention will be considered in consideration of the detailed description that is set forth below in conjunction with the accompanying drawings. And some of which will be apparent to those skilled in the art, or may be made apparent by practice of the invention. The advantages of the invention may be realized and attained by means of the instruments and combinations particularly pointed out in the appended claims.
BEST MODE FOR CARRYING OUT THE INVENTION
[0023]
Specific embodiments of the present invention are described in considerable detail below to illustrate its principles and operation. However, various modifications can be made, and the scope of the present invention is not limited to the specific embodiments described later. For example, while biological systems and assays are specifically mentioned for gene sequencing and DNA hybridization, the methods and systems may be used to fabricate and test optical circuits, fabricate nanocomposites, and fabricate optoelectronics. Used in fields such as testing of electronic components, assembly and testing of holographic data storage matrices, drug analysis, genomic analysis, proteomics analysis, facilitation of combinatorial chemistry, promotion of colloid self-assembly, investigation of non-biological materials, etc. Will be recognized.
[0024]
Certain terms are used herein for convenience and reference, but are not limited to the meaning below. A brief definition is provided below.
[0025]
A. A "beamlet" is a sub-beam of light generated by passing a light beam, such as a collimated output from a laser or light-emitting diode, or other energy source, through a medium that diffracts it into two or more sub-beams. Or mention other energy sources. An example of a beamlet would be a higher order laser beam diffracted by a grating.
[0026]
B, "Phase profile" refers to the phase of light or other energy source in the cross section of a beam or beamlet.
[0027]
C, "Phase patterning" refers to a beam of light or a phase profile changing light, including diffraction, phase modulation, mode shaping, splitting, focusing, splitting, forming, or other methods of manipulating the beam or beamlet. It refers to the patterned phase shift imparted to the beamlets. The method of manipulating the beam or beamlet is not limited to the above.
[0028]
D, "Probe" refers to a biological material or other chemical that selectively binds or reacts with a target. Probes include oligonucleotides, polynucleotides, compounds, proteins, peptides, lipids, polysaccharides, ligands, cells, antibodies, antigens, cell organelles, lipids, blastomeres, cell aggregates, microorganisms, complementary DNA, RNA Etc., but is not limited thereto.
[0029]
E. FIG. "Target" refers to a biological or other chemical substance whose presence or absence in a sample is detected by binding or reacting the target with a probe. For example, the presence of a target formed from the genetic material is indicative of a reaction such as a hybridization reaction between the target genetic material and a probe genetic material having specific properties, ie, having a complementary structure required for hybridization. Is detected by Target materials are also oligonucleotides, polynucleotides, compounds, proteins, lipids, polysaccharides, ligands, cells, antibodies, antigens, organelles of cells, lipids, blastomeres, aggregates of cells, microorganisms, complementary DNA, RNA Etc., but is not limited thereto.
【Example】
[0030]
As shown in FIG. 1, probes 500-504 may bind or react with any suitable substrate by any suitable binding process or protocol. An important feature of a suitable substrate is that it is a material that is contained in and scanned by the light trap. Representative dielectric substrates include beads, irregular small particles, or other regular small particles. Suitable substrates include CPG, ceramics, silica, titanium dioxide, latex, polystyrene, methyl styrene, polymethyl methacrylate, paramagnetic materials, trisol, cross-linked dextrans such as graphite, teflon, sepharose, cellulose, nylon Manufactured from plastic-containing materials such as, but not limited to, cross-linked micelles, liposomes and membrane vesicles.
[0031]
As shown in another embodiment illustrated in FIG. 2, the method of the present invention comprises providing one or more probes 505 (one of which is shown) not bound to a substrate. And one or more optical traps 1005, one of which is shown. It is to be understood that an arrayable array can include only bound probes, only unbound probes, or a combination of bound and unbound probes. When bound and unbound probes are combined, the choice of what combination of bound and unbound probes to take may be influenced in part by the physical properties of the probe. In particular, the properties of certain probes, such as skin cells, can change without contacting the substrate. Conversely, by removing the substrate and maintaining the tertiary structure of the probe / protein, the action of other probes, such as proteins, can be better contributed.
[0032]
FIG. 1 shows an arrayable array 8 of substrate binding probes 500-504 for analysis of biological material. The probe is formed in the test cell 10 using movable light traps 1000-1004 constructed from focused beamlets 2000-2004. The test cell 10 is a container made of a substantially transparent material, which allows the beamlets to penetrate and does not interfere with the formation of the light trap.
[0033]
FIG. 3 shows an overview of a system 20 for creating and changing the position of an array of arrayable probes. Movable optical traps 1000-1004 (FIG. 1) are created in the container 10 by passing a collimated light, preferably a laser beam 100 generated by a laser 102, to an area A ′ of the beam splitter 30. . One of the beams (beam 31) is emitted from laser 102 and is redirected from region A 'of beam splitter 31 to region A on phase patterning optics 22. Each beamlet generated by the phase patterning optics 22 then passes through region B at the back aperture 28 of the focusing lens 12. The beamlets are focused by a focusing lens 12. The focused beamlets form the optical traps 1000-1004 by creating the gradient conditions necessary to encompass and operate the three-dimensional probe. For clarity, only five sets of probes, beamlets and light traps are shown in FIG. 1, but depending on the nature of the analysis, the range and other parameters, and the ability of the system to generate the light traps, It should be understood that more or less can be taken.
[0034]
Any suitable laser can be used as the source of the laser beam 100. Useful lasers include solid state lasers, diode pumped lasers, gas lasers, dye lasers, alexandrite lasers, free electron lasers, VCSEL lasers, diode lasers, titanium sapphire lasers, doped YAG lasers, doped YLF lasers, diode pumped YAG lasers And a flashlamp pumped YAG laser. Diode-pumped Nd: YAG lasers operating between 10 mW and 5 W are preferred. The preferred wavelength of the laser beam 100 used to form the array for the investigation of biological material is infrared, near infrared, red, green, blue, with wavelengths from about 400 nm to about 1060 nm being most preferred. Includes visible light wavelengths.
[0035]
Beam splitter 30 is fabricated from a dichroic mirror, a photon bandgap mirror, an omni-directional mirror, or other similar device. Beam splitter 30 selectively reflects the wavelength of light used to form the optical trap and passes other wavelengths. Thereafter, a portion of the light reflected at the beam splitter area A 'is coupled to the coded phase patterning optics 22 substantially located in a plane 24 adjacent the flat back aperture 28 of the focusing lens 12. It penetrates the area A.
[0036]
When the laser beam 100 is directed by the phase patterning optics 22, the phase patterning optics generates a plurality of beamlets having an altered phase profile. Depending on the number and type of optical traps desired, this modification may include diffraction, wavefront shaping, phase shifting, steering, splitting, and focusing. Based on the selected phase profile, the phase patterning optic is used to create an optical trap in the form of an optical tweezer, optical vortex, optical bottle, optical rotator, optical cage, and a combination of two or more of these. You can do it.
[0037]
In embodiments where the phase profile of the beamlets is not as strong at the edges and becomes stronger from the edges toward the interior region, less than about 15 percent overfilling of the back aperture 28 may be free of overfilling. It is useful to form the light trap with greater intensity at the edges of the light trap compared to the light trap formed by the back aperture 28.
[0038]
Suitable phase patterning optics are characterized as being transmissive or reflective depending on how the focused light beam or other energy source is oriented. Transmissive diffractive optics transmit light or other sources of energy, while reflective diffractive optics reflect beams.
[0039]
Phase patterning optics can also be categorized as having a static or dynamic surface. Examples of suitable static phase patterning optics include diffraction gratings, gratings including reflective and transmissive gratings, holograms including polychromatic holograms, stencils, holographic filters for shaping light, polychromatic holograms, lenses , Mirrors, prisms, waveplates, and the like, including those with one or more fixed surface areas. The static transmissive phase patterning optic 40 is characterized by a fixed surface 41, as shown in FIG. However, in some embodiments, the phase patterning optics are themselves movable, and by moving the phase patterning optics relative to the laser beam to select an appropriate area, the fixed surface areas 42-46. Can be selected. The static phase patterning optics may be mounted on a spindle 47 and rotated by a controlled motor (not shown). The static phase patterning optical element in the embodiment shown in FIG. 4 comprises a fixed surface 41 and individual areas 42 to 46. In other embodiments of the transmissive or reflective static phase patterning optics, the fixed surface 41 may be a non-homogeneous surface that includes a substantially continuously varying region, or a region that varies substantially continuously with an individual region. Have a combination.
[0040]
Examples of suitable dynamic phase-patterning optics that exhibit a time-dependent aspect to their function include computer-generated diffraction patterns, phase-shifting materials, and liquid crystal phase-shifting, including piston-mode micromirror arrays. Includes arrays, spatial light modulators, electro-optic deflectors, acousto-optic modulators, deformable mirrors, reflective MEMS arrays and the like. In dynamic phase-patterning optics, the media containing the phase-patterning optics encodes a hologram, and the hologram can be changed to provide a patterned phase shift to the focused light beam, such as a diffraction or focusing. Resulting in a corresponding change in the phase profile of the light beam. Further, the media can be changed to change the position of the optical trap. This is an advantage of dynamic phase patterning optics, where the media can be changed to move each of the optical traps separately.
[0041]
Suitable dynamic optics are "PAL-SLM Series X7665" manufactured by Hamamatsu Photonics, Japan, or "SLM512N15" and "SLM512SA7" manufactured by Boulder Nonlinear Systems, Lafayette, CO. ”. These phase patterning optics generate beamlets and are controlled to produce beamlets 2000-2004 (FIG. 1) by coded holograms in media that can be changed to select the beamlet format. Computer.
[0042]
In some embodiments, the type of optical trap to form the array and / or the position of the optical trap is changed to allow for alignment and rearrangement. This format can be changed from its original format to the format of optical tweezers, optical vortex, optical bottle, optical rotator or optical cage.
[0043]
In addition, phase patterning optics are useful for imparting a particular phase geometry mode to laser light, for example, by converting from a Gaussian mode to a Gauss-Laguerre mode. Thus, some beamlets may be generated in Gaussian-Laguerre mode and other beamlets may be generated in Gaussian mode.
[0044]
The probe is formed in the container 10. The container 10 is a test cell made of a substantially transparent material, through which the beamlets can penetrate, without interfering with the formation of an optical trap. In those embodiments where the substrate is labeled with a wavelength-specific dye, the test cell should be transparent to the particular wavelength. In addition, the test cell should be made from a material that is inert to the substrate. For example, biological substrates such as cells, proteins and DNA should not stick to the surface of the test cell and should not be altered or destroyed by the cell material.
[0045]
Probes having the particular properties required to bind and / or react with the desired target are selected for addition into a container and inclusion in an arrayable array. In some of the embodiments, where the probe is bound to a substrate, the substrate is labeled with a tracer, such as a wavelength-specific dye, to facilitate probe selection. In a preferred embodiment, all substrate binding probes having the same binding or reaction properties are labeled with the same type of tracer. If the substrate is labeled with a wavelength-specific tracer, selection of probes 500-504 can be accomplished by adding a probe bound to the labeled substrate to vessel 10. Thereafter, measurement of the spectrum of the labeled substrate of the probe, as shown in FIG. 3, can be used to select (or not select) the probes to be included in the array. In some embodiments (FIG. 2), the probe is not bound to a substrate and may be labeled.
[0046]
In embodiments where unlabeled probes are selected to form all or part of an array, the probes can be added to the container 10 in a sequential order. In such cases, the identity of the probes is known by the order of loading. That is, the identity of the probe can be known based on the time at which the probe was added. Alternatively, probes with different binding or reaction properties can be separated into distinct predetermined locations based on the differences in properties. Thereafter, probes are selected based on their location in the container.
[0047]
As seen in FIG. 3, spectroscopy of a sample of biological material can be performed with an imaging light source 39 suitable for either spectroscopy or polarized backscatter. The former is useful for assessing chemical identity, and the latter is suitable for measuring internal structure dimensions, such as nucleus size. In some embodiments, using such spectroscopic methods, cells are removed and an array of probes is created from the selected and removed cells. For example, the computer 38 can be used to analyze the spectral data and identify suspected cancerous cells, pre-cancerous cells, and / or non-cancerous cells. The computer can then apply this information to instruct the light trap to include the selected cell type. The included cells can then be used as analytical probes based on reaction or binding with other cells, antibodies, antigens, or other biological materials, or targets such as drugs and other chemicals. The method used to locate and recruit cells based on parameters specific to cancer cells allows for the retrieval and / or recruitment of blastomeres, cells or other material without departing from the scope of the present invention. It will be appreciated by those skilled in the art that it can be modified to be used for
[0048]
In other embodiments, labeled probes or unlabeled probes, such as unlabeled probes having different binding or reaction properties, are placed in a row of subcells 16 disposed in the container 10. May be placed in In FIG. 1, only one subcell is shown for clarity. However, it should be understood that multiple such sub-cells are provided. In some embodiments, subcell boundaries are constructed with optical traps. A number of correctly oriented light traps create an optical subcell that can perform the same function as the physical subcell 16.
[0049]
The placement of the probes in the subcell 16 is performed by any suitable means, including operation by optical traps, by flow channels, by microcapillaries, or by other similar mechanisms. Within each subcell is located one or more probes having the same binding or reaction characteristics. Thereafter, the selection of a probe to be placed in the array is made based on the subcell in which the probe is contained.
[0050]
The optical traps 1000-1004 are then used to trap the selected probes 500-504 by including the probes in the optical traps 1000-1004. Such included probes are formed to form an array.
[0051]
The method and system of the present invention are suitable for semi-automated or automated processes for tracking the operation of each optical trap and its contents. This operation can be monitored by video cameras, spectral or optical data streams, which control the selection of probes and the type of probes included in the optical trap and the composition of the probes forming the array. Computer control of the generation of useful optical trap information. In another embodiment, this operation is tracked based on a predetermined operation of each optical trap caused by the encoding of the phase patterning optics. Further, in some embodiments, a computer is used to maintain a record of each probe included in each light trap.
[0052]
Referring again to the beam splitter 30, the beam splitter 30 transmits light data corresponding to the position of one or more beamlets derived from the position and location of the probe contained in the optical trap through the test cell 10. A light beam 32 from an imaging light source 39 that forms a stream is provided.
[0053]
The optical data stream is then viewed, converted to a video signal, monitored, or analyzed by a visual inspection 34a with an operator 36, a spectrograph 34b and / or a video monitor 34c. The optical data stream 32 is processed by a photodetector 34d for monitoring intensity or any suitable device that converts the optical data stream into a digital data stream for use by a computer 38.
[0054]
To build the array, an operator 36 and / or computer 38 was coded by the phase patterning optics 22 to command the operation of each light trap to obtain and trap the selected probe. Will adjust the hologram. The plurality of light traps containing the probes form an arrayable array of compositions that can be reconfigured with respect to probe composition and location depending on the needs of the user. Using the optical data stream, the location of one or more trapped probes can be identified and these locations are monitored. In each of some embodiments, based on such information, the surface of the phase patterning optic can be changed to change the shape of one or more light traps containing the probe.
[0055]
Further, the position of one or more probes trapped in the array can be tracked by monitoring the position of the optical trap containing the probe. Then, using such information, any probes in the array may be separately repositioned in the test cell by altering the surface of the phase patterning optics, and the identity of each probe is Regardless of where the trap locates the probe, it is stored by the optical trap in which the probe is contained.
[0056]
In a preferred embodiment, computer 38 controls the operation of the optical trap before and after the probe is trapped. In another embodiment, the optical data stream is first converted to a video signal, which is then used to generate an image corresponding to the array, and the operator can operate the at least one optical trap based on the image. Look at the image to control.
[0057]
Referring to FIGS. 1 and 3, a batch of first targets T1-T5 is added through inlet 14 to a test cell 10 containing a fluid medium 3000 therein for performing an analysis. The array of probes 500-504 is retained in culture medium 3000 by containment by optical traps 1000-1004. In order to increase the chance of interaction with the targets T1 to T5, the probe may be moved with respect to the test cell in response to the operation of the optical trap.
[0058]
For example, in one embodiment, probes 500-504 are rolled in medium 3000 containing targets T1-T5. By including the probe optically rather than physically, moving the probe within the test cell 10 increases the chance of interaction between each target and the probe, thus increasing the speed and efficiency of the analysis. Is improved.
[0059]
The operation of an array of probes 500-502 by sequentially generating multiple optical traps is shown in FIG. In the embodiment shown in FIG. 5 (a), a simple linear movement of an array of probes formed along a line P1 representing a first predetermined position is shown. The operation is performed by transferring the probe from a first predetermined position of the optical trap to a second, third and fourth position. Still referring to FIG. 4, a first group of first light traps is created by directing a laser beam to a first region 42 of the phase patterning optic 40. As the beamlets emitted from the first region 42 pass through the focusing lens, the beamlets form a first group of optical traps at a first position P1 that includes the probes 500-503.
[0060]
In order to move the probes 500 to 502 from the first position P1 to the second position P2, the laser beam is aligned with the second region 43 for generating the second optical trap group corresponding to the second predetermined position P2. To do so, the static phase patterning optic 40 rotates about a spindle 47. By constructing the second group of optical traps sufficiently near the first position P1, the probe can be moved from the first group of optical traps to the second group of optical traps. The sequence moves the probe from the second predetermined position P2 to the third predetermined position P3 by rotating the phase patterning optics to align the appropriate regions 42-46 corresponding to the desired positions P1-P5. It may move continuously from the third predetermined position P3 to the fourth predetermined position P4 and from the fourth predetermined position P4 to the fifth predetermined position P5. The time interval between the end time of one set of optical traps and the start time of the next set of optical traps is a time interval that ensures that the probe can move to the next set of optical traps without drifting out. Should be.
[0061]
Such operation of the probe is useful to increase the chance of the probe rolling in the medium, thereby increasing the chance of the target in the medium interacting with the probe. This kind of simple movement may also be useful in moving the subcell 16 (FIG. 1) to another area within the test cell 10 or separating the probe from the subcell 16.
[0062]
FIG. 5B shows a staggered movement of the probe from a wide interval to a narrow interval in the embodiment shown in FIG. The staggered movement of the probe occurs in a manner similar to that described with respect to FIG. However, the first area 42 includes two probes 500 and 502 formed along the line P1 and a third probe 501 located at P2 which is intermediate between the two probes but is separated from the line P1. A staggered optical trap group is generated. The staggered arrangement of the probes allows the probes to move from the first group of optical traps to the second group of optical traps and to be densely packed as they move to the second and subsequent positions. This prevents the trap group from approaching too close to the two probes at the same time and prevents the probes from being included in the wrong optical trap.
[0063]
Once the target has interacted with the probe, spectral methods can be used to probe the target. The spectrum of the probe with a positive result (ie, the probe reacts with or binds to the target) can be obtained by using an imaging light source 39 suitable for either inelastic spectroscopy or polarized backscattering. Computer 38 can analyze the spectral data to identify desired targets and to instruct phase patterning optics to separate those desired targets. The method used to separate the targets based on the spectral data can be used to identify and target the targets based on other information obtained from the optical data stream without departing from the scope of the present invention. It will be appreciated by those skilled in the art that (or) it can be modified to be used for separation.
[0064]
After the analysis is completed, a selection is made by computer 38 and / or operator 36 as to which probes to discard and which probes to collect. The array can be rearranged so that a given optical trap and the probes contained therein can be moved selectively. In some cases, medium 3000 and unbound target are removed or washed away from test cell 10 via outlet 18 and the analysis is terminated. In other cases, at least some of the probes still included in the optical trap are reused with additional targets for further analysis. This technique can be useful in the case of probes that test positive or negative, depending on the parameters of the analysis. In yet another case, the light trap discards unbound probes and replaces additional probes for further experimentation, as the array of probes can be rearranged depending on the amount and properties of the probes forming the array. Can be used to obtain.
[0065]
In some embodiments, it is not necessary to generate a beamlet from each region of the static beam changing optical member 40 or to move the static beam changing optical member 40 to a predetermined position. There is no. Instead, changing the order of the regions will change the location of the light traps.
[0066]
An elevation view of a compact system for forming the light trap 50 is shown in FIG. The phase patterning optical member 51 is manufactured by "Hamamatsu Photonics, Japan, manufactured by PAL-SLM Series X7665", or "SLM512SA7" and "SLM512SA15" manufactured by Boulder Nonlinear Systems, Inc. of Lafayette, CO. A dynamic optical element that is a parallel-aligned spatial light modulator with a reflective and dynamic surface, such as
[0067]
FIG. 6 (a) shows a compact system for forming an optical trap. The optical member 51 is aligned or connected with the housing 52, through which a first optical channel 53a is provided. One end 53b of the first optical channel is immediately adjacent to the optical member 51, and the other end 53c of the first optical channel intersects and communicates with the orthogonal second optical channel 53d. A second light channel is formed in the base 54a of the turret or "revolver" 54b for mounting the microscope lens. The revolver 54b is mounted on a TE200 microscope (not shown) manufactured by Nikon Corporation. The second optical channel is further in communication with a third optical channel 55a orthogonal to the second optical channel. The third light channel 55a traverses from the upper surface of the revolver 54b to the base 54a of the revolver and is parallel to the objective lens focusing lens 56. The focusing lens has a top and a bottom forming a back aperture 57. A dichroic mirror beam splitter 58 is located in the third optical channel, between the second optical channel and the back aperture 57 of the focusing lens. Other components in the compact system for forming the optical trap 50 are a first mirror M1, which reflects a beamlet radiating from the phase patterning optics to the first optical channel, and which is reflected by the first mirror M1. A first transmission optics TO1 aligned to receive the beamlets and disposed in the first optical channel; and a first transmission optics TO1 aligned to receive the beamlets passing through the first transmission lens TO1 and disposed in the first optical channel. A second transmission optical system TO2 and a second mirror positioned to reflect beamlets passing through the second transmission optical system TO2 and the third optical channel 55a and located at the intersection of the first optical channel and the second optical channel. M2.
[0068]
A laser beam (not shown) is directed through optics 150 external to collimator end 151 and reflected by dynamic surface 59 of optical member 51 to create an optical trap. Light (not shown) that excites the collimator end 151 of the optical fiber 150 is diffracted by the dynamic surface 59 of the optical member 51 into a plurality of beamlets (not shown). The number, type, and orientation of each beamlet can be controlled and changed by changing the hologram encoded in the media on the dynamic surface 59. Thereafter, the beamlet is reflected by the first mirror M1, goes down the first optical channel 53a through the first transmission optical system TO1, reaches the second mirror M2 through the second transmission optical system TO2, and becomes a dichroic mirror. It is directed by 58 to the back aperture 57 of the objective lens 56 and is focused by the objective lens 56 where the light gradient required to form the light trap is created. A portion of the light for imaging, which separates through dichroic mirror 58, passes through a lower portion of third optical channel 55b to form an optical data stream (not shown).
[0069]
In embodiments where the phase profile of the beamlets is not as strong at the edges and becomes stronger from the edges toward the interior region, overfilling of the back aperture 57 of less than about 15 percent will result in no overfilling. It is useful for forming an optical trap with greater intensity at the edge of the optical trap as compared to the optical trap formed by the back aperture 57.
[0070]
An elevation view of a Nikon TE200 microscope 60 mounted with a compact system for forming the optical trap 50 is shown in FIG. 6 (b). A revolver 54 attached to the housing 52 is mounted directly to the microscope by mounts (not shown) for the revolvers 54a and 54b. The housing and its contents and the attached optics 51 are fixed to the revolvers 54a and 54b, with little or no change required for the rest of the microscope. A light source 61 may be provided on top of the objective lens 56 for imaging.
[0071]
The first transmission optical system TO1 and the second transmission optical system TO2 are each shown including two lens members. The lens can be either a convex lens or a concave lens. To achieve image transfer from the first mirror M1 to the second mirror M2, a symmetric singlet separated by air, a symmetric doublet separated by air, and / or an additional lens or lens group Different different types and numbers of lenses can be selected. In some embodiments, the first and second transmission optics are double lines separated by air and are slightly separated to also function as a telephoto lens.
[0072]
Since certain modifications can be made to the above system devices and methods within the scope of the present invention in connection with the present specification, all matter contained in the above description is illustrated as illustrated in the accompanying drawings and specification. It is intended to be interpreted as non-limiting.
[Brief description of the drawings]
[0073]
FIG. 1 is a cutaway side view of a system for forming an array of probes that can be arranged.
FIG. 2 shows a free probe contained within an optical trap.
FIG. 3 shows an overview of a system for forming an array of probes.
FIG. 4 shows a beam changing member with a plurality of static regions.
5A illustrates a first operation of the probe, and FIG. 5B illustrates a second operation of the probe.
FIG. 6 (a) is a block diagram of a compact system for forming an optical trap.
6 (b) shows an inverted microscope to which the compact system of FIG. 6 (a) is connected.
[Explanation of symbols]
[0074]
8 ... arrayable array,
10 ... test cell (container),
12 ... Focusing lens,
14. Entrance
16 ... Subcell
18 ... Exit
22 ... phase patterning optical member,
28 ... back aperture,
100 ... laser beam,
102 ... Laser,
500-504 ... substrate binding probe,
2000-2004 ... beamlet,
1000 to 1004 ... light trap,

Claims (157)

A method of arranging and tracking probes, comprising:
Creating at least two movable light traps in the container;
Providing at least two probes in the container,
Selecting at least two probes for inclusion in the probe array contained in the optical trap;
Trapping each of said selected probes into one of said optical traps to arrange a probe array contained within said optical trap;
Tracking the position of at least one trapped probe by monitoring the position of the optical trap containing the trapped probe. The method of claim 1, further comprising changing a position of at least one tracked probe by moving an optical trap containing the tracked probe. The method of claim 1, wherein the optical trap comprises two or more optical tweezers, optical vortex, optical bottle, optical rotator or optical cage. The method of claim 2, wherein each of the optical traps is independently movable. 3. The method of claim 2, wherein the operation of each light trap is controlled by a computer. The method of claim 4, wherein the operation of each light trap is controlled by a computer. At least one of the probes is bound to a substrate labeled with a specific wavelength marker, and at least one of the bound probes measures the marker with a spectrometer and uses the spectrometer measurement. 5. The method according to claim 4, wherein the selection is made by selecting. At least two of the probes have different binding or reaction characteristics from each other, and at least one probe moves the probe to a predetermined position in the container based on its different binding or reaction characteristics; The method according to claim 4, wherein the selection is performed by separating the probes by a method of selecting using the positions of the separated probes. The method of claim 8, wherein the predetermined location is a physical subcell. The method of claim 8, wherein the predetermined location is an optical subcell. 2. The method of claim 1, further comprising introducing at least one target into the container and measuring whether there is a reaction between each target and each trapped probe. The method of claim 11, wherein the trapped probe is a biological material. The method of claim 11, wherein the trapped probe is a compound. The method of claim 12, wherein the target is a biological material. The method of claim 12, wherein the target is a compound. 14. The method of claim 13, wherein the target is a biological material. 14. The method of claim 13, wherein said target is a compound. The trapped probes may be oligonucleotides, polynucleotides, proteins, polysaccharides, ligands, cells, antibodies, antigens, cytoplasmic organelles, lipids, blastomeres, aggregates of cells, microorganisms, peptides, complementary DNA, RNA or 13. The method of claim 12, wherein the combination is a combination thereof. The target is an oligonucleotide, a polynucleotide, a protein, a polysaccharide, a ligand, a cell, an antibody, an antigen, a cytoplasmic organelle, a lipid, a blastomere, a cell aggregate, a microorganism, a peptide, a complementary DNA, an RNA or a combination thereof. The method of claim 14, wherein The target is an oligonucleotide, a polynucleotide, a protein, a polysaccharide, a ligand, a cell, an antibody, an antigen, a cytoplasmic organelle, a lipid, a blastomere, a cell aggregate, a microorganism, a peptide, a complementary DNA, an RNA or a combination thereof. 17. The method of claim 16, wherein the method is selected from one or more groups consisting of: The method of claim 1, wherein all of said probes bind to a substrate. The method of claim 1 wherein all of said probes are directly trapped by said optical trap. 2. The method of claim 1, wherein at least some probes bind to the substrate and at least some probes do not bind to the substrate. 22. The method of claim 21, further comprising changing a position of at least two probes tracked in the array by moving an optical trap containing the probes. The method of claim 1, further comprising generating an optical data stream of data corresponding to the identity and location of at least one optical trap. The method of claim 24, wherein each light trap is independently movable. The method of claim 24, wherein the operation of each light trap is controlled by a computer. The method of claim 25, further comprising receiving the optical data stream using a computer. The method of claim 28, further comprising analyzing the optical data stream using a computer. The method of claim 29, wherein the computer commands operation of at least one optical trap based on analysis of the optical data stream. The method of claim 25, further comprising converting the optical data stream to a video signal. The method of claim 31, further comprising receiving the video signal using a computer. The method of claim 32, further comprising analyzing the video signal with the computer. 34. The method of claim 33, further comprising using the computer to command operation of one or more optical traps based on the analysis of the video signal. The method of claim 31, wherein the video signal is used to form an image. The method of claim 35, further comprising: an operator viewing the image and instructing operation of one or more light traps based on viewing the image. The method of claim 25, wherein the data is spectral data. 38. The method of claim 37, further comprising using a computer to command operation of one or more optical traps based on the analysis of the spectral data. The method of claim 24, wherein the optical trap comprises at least two of optical tweezers, optical vortex, optical bottle, optical rotator, or optical cage. 27. The method of claim 26, wherein the operation of each light trap is controlled by a computer. The method of claim 24, wherein the at least one probe is selected by measuring a marker with a spectrometer and selecting the at least one probe using the spectrometer measurement. At least two of the probes have different binding or reaction characteristics from each other, and at least one probe moves the probe to a predetermined position in the container based on its different binding or reaction characteristics; The method according to claim 24, wherein the selection is performed by separating the probes by a method of selecting using the positions of the separated probes. 43. The method of claim 42, wherein said predetermined location is a physical subcell. 43. The method of claim 42, wherein said predetermined location is an optical subcell. 169. The method of claim 169, wherein said trapped probe is a biological material. 169. The method of claim 169, wherein said trapped probe is a compound. 47. The method of claim 46, wherein said target is a biological material. 47. The method of claim 46, wherein said target is a compound. The method of claim 45, wherein the target is a biological material. The method of claim 45, wherein the target is a compound. The trapped probes may be oligonucleotides, polynucleotides, proteins, polysaccharides, ligands, cells, antibodies, antigens, cytoplasmic organelles, lipids, blastomeres, aggregates of cells, microorganisms, peptides, complementary DNA, RNA or The method of claim 45, wherein the combination is a combination thereof. The target is an oligonucleotide, a polynucleotide, a protein, a polysaccharide, a ligand, a cell, an antibody, an antigen, a cytoplasmic organelle, a lipid, a blastomere, a cell aggregate, a microorganism, a peptide, a complementary DNA, an RNA or a combination thereof. 48. The method of claim 47, wherein The target is an oligonucleotide, a polynucleotide, a protein, a polysaccharide, a ligand, a cell, an antibody, an antigen, a cytoplasmic organelle, a lipid, a blastomere, a cell aggregate, a microorganism, a peptide, a complementary DNA, an RNA or a combination thereof. 50. The method of claim 49, wherein the method is selected from one or more groups consisting of: 25. The method of claim 24, wherein all of said probes are bound to a substrate. 25. The method of claim 24, wherein all of said probes are unbound to the substrate. 25. The method of claim 24, wherein at least some of the probes are bound to the substrate, and at least some of the probes are unbound to the substrate. A method for analyzing biological material, comprising:
Creating at least two movable light traps in the container;
Providing a fluid medium in the container,
Providing at least two biological substance probes in the fluid medium,
Selecting at least two probes for inclusion in the array;
Trapping each of said selected probes into one of said optical traps;
Introducing at least one target containing the biological material into the container,
Determining whether there is a reaction between each of the trapped probes and each of the targets. 58. The method of claim 57, further comprising tracking a position of at least one trapped probe by monitoring a position of the light trap containing the trapped probe. 58. The method of claim 57, wherein the trapped probe comprises a biological material. 58. The method of claim 57, wherein said trapped probe comprises a compound. The trapped probes may be oligonucleotides, polynucleotides, proteins, polysaccharides, ligands, cells, antibodies, antigens, cytoplasmic organelles, lipids, blastomeres, aggregates of cells, microorganisms, peptides, complementary DNA, RNA or The method of claim 59, wherein the combination is a combination thereof. The target is an oligonucleotide, a polynucleotide, a protein, a polysaccharide, a ligand, a cell, an antibody, an antigen, a cytoplasmic organelle, a lipid, a blastomere, a cell aggregate, a microorganism, a peptide, a complementary DNA, an RNA or a combination thereof. 58. The method of claim 57, wherein The method of claim 57, further comprising generating an optical data stream of data corresponding to the identity and location of at least one optical trap. 64. The method of claim 63, further comprising: changing a position of at least one trapped probe in the array by moving an optical trap containing the probe. 65. The method of claim 64, wherein each light trap is independently movable. The method of claim 64, wherein the operation of each light trap is controlled by a computer. The method of claim 63, further comprising receiving the optical data stream using a computer. The method of claim 67, further comprising analyzing the optical data stream using a computer. 70. The method of claim 68, further comprising using the computer to command operation of one or more optical traps based on analysis of the optical data stream. The method of claim 63, further comprising converting the optical data stream to a video signal. The method of claim 70, further comprising receiving the video signal using a computer. The method of claim 71, further comprising analyzing the video signal using the computer. The method of claim 72, further comprising using the computer to command operation of one or more optical traps based on analysis of the video signal. The method of claim 70, wherein the video signal is used to generate an image. 75. The method of claim 74, further comprising an operator viewing the image and instructing operation of one or more light traps based on viewing the image. The method of claim 63, wherein said data is spectral data. 77. The method of claim 76, further comprising using a computer to direct the operation of one or more optical traps based on the analysis of the spectral data. 64. The method of claim 63, wherein said optical trap comprises at least two of optical tweezers, optical vortex, optical bottle, optical rotator, and optical cage. 64. The method of claim 63, wherein at least one of said probes is bound to a substrate. 64. The method of claim 63, wherein at least one of said probes is unbound to a substrate. 80. The method of claim 79, wherein all substrate binding probes having the same binding or reaction properties are labeled with the same label. The method of claim 81, wherein at least one of said labels is a wavelength-specific dye. At least one of said substrate binding probes is selected by measuring a spectral response of said wavelength-specific dye and using a spectral measurement to select at least one of said substrate binding probes. Clause 82. The method of clause 82. At least two of the probes have different binding or reaction characteristics from each other, and at least one of the probes separates the probes based on different binding or reaction characteristics, and moves the probe to a predetermined position in the container. 64. The method of claim 63, wherein the selected probe is selected by moving to a location and using the location of the separated probe to select one of the probes. The method of claim 63, wherein the predetermined location is a physical subcell. The method of claim 84, wherein the predetermined location is an optical subcell. A method of arranging probe arrays, comprising:
Creating at least two movable light traps in the container,
Providing at least two of said probes in said container and arranging said at least two probe arrays by selecting each probe with one of said optical traps. A method of arranging probe arrays, comprising:
Directing a focused beam of light to the phase patterning optical member to form a plurality of beamlets radiated from the phase patterning optical member;
Passing the plurality of beamlets through a focusing lens to create a movable optical trap in the container, and a back aperture to converge the plurality of beamlets emitted from the focusing lens. Directing the plurality of beamlets,
Supplying a plurality of probes in the container,
Selecting at least two probes for inclusions in the probe array contained within the optical trap;
Trapping each of the selected probes with one of the optical traps to arrange a probe array contained within the optical trap, and rearranging the probe array contained within the optical trap; Moving the optical trap containing the probe to change the position of at least one of the probes contained in the optical trap. The method of claim 90, wherein the phase patterning optic has a static surface. The method of claim 91, wherein the static surface comprises two or more discrete regions. 90. The method of claim 90, wherein the position of at least one probe contained within the light trap is changed by changing the discrete area of the static surface at which the beam of light is directed. . 90. The method of claim 89, wherein said static surface changes substantially continuously. 90. The method of claim 89, wherein the position of the at least one light trap is changed by changing the area of the static surface where the beam of light is directed. 90. The method of claim 89, wherein said beam changing optic is a grating, hologram, stencil, light shaping holographic filter, lens, mirror, prism or wave plate. 91. The method of claim 90, wherein each of said discrete regions is a grating, hologram, stencil, light shaping holographic filter, lens, mirror, prism or wave plate. The method of claim 88, wherein the phase patterning optic is dynamic. 97. The method of claim 96, wherein at least one position of a probe contained in said optical trap is changed by changing said dynamic phase patterning optics. 100. The method of claim 97, wherein at least one form of said light trap is changed by changing said dynamic phase patterning optics. The method of claim 97, wherein the altered light trap is an optical tweezer, an optical vortex, an optical bottle, an optical rotator, or an optical cage. 92. The method of claim 91, wherein at least one form of the light trap is varied by moving the individual static surface. The method of claim 100, wherein the altered light trap is an optical tweezer, an optical vortex, an optical bottle, an optical rotator, or an optical cage. 100. The method of claim 97, wherein said dynamic phase patterning optics change is a change in a surface encoded hologram. A system for forming and tracking an optical trap containing a probe, the system comprising:
A light source for producing a focused beam of light;
A substantially transparent container,
An imaging light source for generating a beam of light illuminating the contents of the container;
A beam splitter for pointing,
A phase patterning optical member having a surface for receiving a focused beam of light emitted from the light source and diffracting the beamlet into at least two beamlets and directing each of the beamlets to a back aperture of a focusing lens; Is varied to change at least one phase shape and / or orientation of the beamlets, and wherein the focusing lens focuses each of the beamlets to form an optical trap for containing a probe. A phase patterning optical member,
A monitor for receiving a beam of light illuminating the contents of the container and tracking operation and contents of at least one light trap. 114. The system of claim 103, wherein said container has an inlet. 114. The system of claim 103, wherein said container has an outlet. The method of claim 8, wherein the probes are separated by using an optical trap, flow channel, or capillary action. 43. The method of claim 42, wherein the probes are separated by using an optical trap, flow channel, or capillary action. 85. The method of claim 84, wherein said probes are separated by using an optical trap, flow channel, or capillary action. The target comprises oligonucleotides, polynucleotides, proteins, polysaccharides, ligands, cells, antibodies, antigens, organelles of cells, lipids, blastomeres, aggregates of cells, microorganisms, peptides, complementary DNA, and RNA. The method of claim 63, wherein the method is selected from one or more groups. The method of claim 9, wherein the probes are separated by using a light trap, flow channel, or capillary action. Wherein the phase patterning optics are dynamic and further maintain a first computer for controlling diffraction by the phase patterning optics and a record of each probe contained in each light trap. 104. The system of claim 103, comprising: a first computer. The method of claim 2, wherein the movement of the trapped probe is tracked based on a predetermined movement of each of the optical traps caused by encoding the phase patterning optics. A system for forming and tracking an optical trap containing an optical probe coupled to a target, the system comprising:
A plurality of probes bound to the target;
A light source for producing a focused beam of light;
A substantially transparent container,
An imaging light source for generating a beam of light illuminating the contents of the container;
A beam splitter for directing a beam of the focused light emitted from the light source, and a beam of light for irradiating the contents of the container,
A phase patterning optical member having a surface for receiving a focused beam of light emitted from the light source and diffracting the beamlet into at least two beamlets and directing each of the beamlets to a back aperture of a focusing lens; Is varied to change at least one phase shape and / or orientation of the beamlets, and the focusing lens is coupled to each of the beamlets to form an optical trap containing the probe coupled to the target. Wherein the phase patterning optics and a monitor for receiving a beam of light illuminating the contents of the container and tracking the operation and contents of at least one light trap. Having system. The system of claim 113, wherein the probe is a biological material. The system of claim 113, wherein the probe is a compound. The system of claim 114, wherein the target is a biological material. 115. The system of claim 114, wherein said target is a compound. The system of claim 115, wherein the target is a biological material. The system of claim 115, wherein the target is a compound. 115. The system of claim 114, wherein said probes are selected from one or more groups consisting of oligonucleotides, polynucleotides, proteins, peptides, complementary DNA, and RNA. The target comprises oligonucleotides, polynucleotides, proteins, polysaccharides, ligands, cells, antibodies, antigens, organelles of cells, lipids, blastomeres, aggregates of cells, microorganisms, peptides, complementary DNA, and RNA. The system of claim 116, wherein the system is selected from one or more groups. The target comprises oligonucleotides, polynucleotides, proteins, polysaccharides, ligands, cells, antibodies, antigens, organelles of cells, lipids, blastomeres, aggregates of cells, microorganisms, peptides, complementary DNA, and RNA. 119. The system of claim 118, wherein the system is selected from one or more groups. At least one light and wrap movement is selected from one or more groups consisting of rotation in a fixed position, rotation in a non-fixed position, movement in two dimensions, and movement in three dimensions. 3. The method of claim 2, wherein 3. The method of claim 2, further comprising moving the optical trap containing the tracked probe by changing the surface of the phase patterning optic. 114. The system of claim 103, wherein the phase patterning optic has a static surface. The system of claim 125, wherein said static surface comprises two or more distinct regions. 129. The system of claim 126, wherein the static surface is movable to focus a focused beam of light on a selected area of the static surface. The phase patterning optic has a static surface with two or more discrete areas, and the location of at least one light trap defines the discrete area of the static surface where a beam of light is directed. 3. The method of claim 2, wherein the change is varied. 114. The system of claim 103, wherein the phase patterning optic has a substantially continuously varying static surface. 130. The system of claim 127, wherein the phase patterning optic is selected from the group consisting of a grating, hologram, stencil, photoforming holographic filter, lens, mirror, prism or wave plate. 127. The system of claim 126, wherein each of said discrete regions is a grating, hologram, stencil, light shaping holographic filter, lens, mirror, prism or waveplate. The system of claim 103, wherein said phase patterning optic is dynamic. The method of claim 2, wherein the phase patterning dynamic member is dynamic, and changing the phase patterning optic changes the position of at least one optical trap. 5. The method of claim 4, wherein the phase patterning dynamic member is dynamic and changing the phase patterning optic changes the position of at least one optical trap. The phase patterning dynamic member is dynamic and changing the phase patterning optical member changes at least one form of the optical trap to an optical tweezer, optical vortex, optical bottle, optical rotator, or optical cage. 3. The method of claim 2, wherein: The phase patterning optic has a static surface with two or more discrete regions, and at least one form of the light trap is changed by moving the static surface. 3. The method of claim 2, wherein 137. The method of claim 136, wherein the configuration of the altered optical trap is selected from the group consisting of optical tweezers, optical vortex, optical bottle, optical rotator, and optical cage. The phase patterning optics may include computer generated diffraction patterns, phase shifting materials, liquid crystal phase shifting arrays including piston mode micromirror arrays, spatial light modulators, electro-optic deflectors, acousto-optic modulators, deformable 133. The system of claim 132, wherein the system is selected from at least one of the group consisting of: a mirror, a reflective MEMS array array. 133. The system of claim 132, further comprising a computer for controlling said dynamic phase patterning optics. 114. The system of claim 103, further comprising a subcell in said container for separating at least one of the optical traps containing probes. 141. The system of claim 140, wherein said subcell is a physical subcell. 163. The system of claim 150, further comprising a computer that changes the phase patterning optics to change the orientation of at least one of the beamlets and moves a corresponding light trap to contain the probe. The system of claim 103, wherein the light source is a laser for producing a focused beam having a wavelength in the green spectrum. The system of claim 103, wherein the light source is a laser for producing a focused beam having a wavelength in the visible blue spectrum. The system of claim 103, wherein the light source is a laser for producing a focused beam having a wavelength in the visible red spectrum. The system of claim 103, wherein the light source is a laser for generating a focused beam having a wavelength between about 400 nm and 1060 nm. The system of claim 103, wherein said light source is a laser beam. 104. The system of claim 103, further comprising a computer for receiving the optical data stream. An apparatus for forming an array of optical traps, comprising:
A light source for producing a focused beam of light;
A focusing lens having a top and a bottom, wherein the bottom receives a focused beam of light emitted from the light source and a focusing lens, wherein the bottom forms a back aperture, and diffracts into at least two beamlets; A phase patterning optical member having a surface for directing each of the beamlets to a back aperture of a focusing lens, and a first optical channel having a first end and a second end, wherein the first end is the phase patterning optical member. A first optical channel characterized by communicating with the member;
A second optical channel having a first end and a second end, wherein the second end crosses the second end of the first optical channel;
A third optical channel having a first end and a second end, a fourth optical channel having the first end communicating with a second end of the second optical channel;
A first mirror for reflecting the beamlet emitted from the phase patterning optical member via the first optical channel;
A first set of transport optics disposed in the first optical channel, the first set of transport optics being arranged to receive the beamlets reflected by the first mirror. When,
A second set of transport optics disposed in the first optical channel, the second set of transport optics being arranged to receive the beamlets passing through the first set of transport lenses. Set and
A second mirror positioned at the intersection of the first light channel and the second light channel to reflect beamlets passing through the second set of transport optics through the third light channel. A second mirror characterized by being aligned;
A third mirror disposed within the third light channel to reflect a beamlet passing through the third light channel to the back aperture of the focusing lens and to form an array of light traps; An apparatus having 149. The apparatus of claim 149, further comprising an illumination source disposed adjacent the top of the focusing lens for generating a beam of illumination light. 160. The apparatus of claim 150, wherein the third mirror is a dichroic beam splitter for directing a beam of focused light emanating from the light source and a beam of light emanating from the illumination source. 150. The apparatus of claim 149, wherein each set of the transport optics is selected from the group consisting of a symmetric single line separated by air, and a symmetric double line separated by air. 149. The apparatus of claim 149, wherein each set of transport optics is selected from the group consisting of a convex lens and a concave lens. The first and second sets of transport optics are air-spaced symmetrical singlets, and are spaced apart at a distance to operate as a telephoto lens in combination. 149. The apparatus of claim 149. 26. The method of claim 25, further comprising incorporating at least one target into the container and determining a response or deficiency of each of the trapped probes comprising each of the targets. The method of claim 1, further comprising moving at least one of the trapped probes by transporting the probe from one optical trap to another. The method of claim 1, further comprising moving at least three of said probes wrapped therewith by transporting said probes from a first set of light traps to a second set of light traps.

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