CN1545561A - Configurable dynamic three-dimensional array - Google Patents

Abstract

The present invention relates generally to configurable probe arrays for analyzing targets in fluids. These probes are contained in optical traps, which allows for varying the quantity and quality of probes in the selection and reconfiguration array. Furthermore, the array is dynamic in that once configured, the optical traps can allow a given optical trap and contained probe to be repositioned independently.

Description

Configurable Dynamic 3D Array

background of the invention

Throughout the application various publications are referenced in parentheses. In order to more fully describe the state of the art to which the present invention pertains, the entire disclosure content of these published documents is incorporated by reference in this application.

1. Field of Invention

The present invention generally relates to probe arrays. In particular, the present invention relates to systems and methods that use multiple optical traps to form a configurable dynamic array of probes that may or may not be bound to a substrate.

2. Discussion on related technical fields

Arrays of promising reactive probes have a long history of use in analytical 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 of biological or chemical material (called targets). The samples analyzed are often only available in relatively small quantities. This limited supply of some materials has led to the development of microarrays, which are used to provide a relatively high density of probes in a small array for target analysis in small numbers of samples.

Microarrays used in the testing of biological materials are often referred to as biochips. Two major applications of biochips are: the extraction of sequence information about a particular nucleic acid, i.e., whether the nucleic acid corresponds to the entire genome of an organism, a single gene, or a portion of a single gene (US Pat. No. 6,025,136 ); and assessment of gene expression. (see Schena, Ad. et al. "Quantitative monitoring of gene expression patterns with a complimentary DNA microarray", Science 270(5235): 467-70 (Oct.20, 1995); D.J. and Winzeler, E.A., "Genomics, gene expression and DNA arrays", Nature 405(6788): 827-836 (2000) and Ekins, R. and Chu, F. <RTI W., "Microarrays: their origins and applications", Trends in Biotechnology 17: 217-18 (1999) .)

Conventional microarrays consist of either linear or two-dimensional structures of oligonucleotide probes attached to the flat surface of a solid support (substrate). Different types of oligonucleotides are immobilized on the substrate in predetermined positions. Thus, once the microarray is formed, the positions of the probes and thus any targets with which the probes react are always known. Immobilization of probes is achieved either by direct synthesis of oligonucleotides onto the substrate by a process known as in situ photolithography synthesis (US Pat. Nos. 5,837,832 and 5,143,854), or by It is achieved by immobilizing oligonucleotides after synthesis.

A disadvantage of such microarrays is that their linear or two-dimensional structure provides a limited surface area on which probes can be immobilized, thereby setting a limit on the density of probes for the assay target. In the case of DNA hybridization between a target (DNA or DNA fragment) and a probe (an immobilized oligonucleotide), the rate of hybridization is controlled by the rate at which the target can come into contact with the probe. Thus, the higher the density of probes, the greater the rate of hybridization.

A second disadvantage of such microarrays arises from the method of their construction. Once the microarray is prepared, the type and number of probes are fixed.

In another method for analyzing targets in small samples, probes are immobilized on the surface of a bead-like substrate. (Pending patent WO00/61198 to Kambara & Mitsuhashi.) Each bead containing a different probe is labeled with a different label, thus, after the analysis is completed, it is allowed to identify which beads have what label. Each probe and associated target are identified (see WO 00/71243).

By physically moving the beads while the probes are secured in the guides, capillaries, grooves, or wells in the sheet, and then washing the beads with the target, the identity of the beads and probes is maintained. While the non-planar nature of the beads does provide a larger surface area for targets to interact with than microarray probes, the beads still must remain in a predetermined order throughout the assay to maintain which probe is supported by the bead. or beads to which the probe must be attached, and a record of the identity of each bead that is checked after analysis to determine its identity.

Another disadvantage of microarray and bead assays is the requirement for the probes to be physically immobilized on the substrate. In some cases, this immobilization itself and spontaneously alters the probe, or affects the process by which the probe is being used for analysis. In other cases, during or after the initial analysis, if the identity of the probes is known throughout the analysis, and the structure of the array can be easily changed, information can be obtained to make desired changes in the quantity and quality of the probes. However, this modification is not possible regardless of microarray or bead analysis.

In an unrelated technical field, it is known to optically trap particles with multiple simultaneously generated optical tweezers. (See US Patent US 6,055,106 issued to Grier & Dufresne) Optical tweezers use the gradient force of a beam of light to trap particles based on their dielectric constant. To minimize energy, particles with a higher dielectric constant than the surrounding medium move to the region of the optical tweezers, where the electric field strength is highest.

Other types of traps that can be used to optically trap particles include, but are not limited to, optical vortices, optical bottlenecks, optical rotators, and light cages. Optical vortices create a surrounding zero The gradient of the area of the electric field used to control particles with a lower permittivity than the surrounding medium, or reflective particles, or other types of particles that are repelled by optical tweezers. To minimize their energy, the particle moves to the region of lowest electric field strength, the region of zero electric field at the focal point of an appropriately shaped laser beam. The optical vortex provides a region of zero electric field much like a hole in a donut (annular chamber). The optical gradient is radial, with the highest electric field at the circumference of the donut. The optical vortex traps a small particle in the hole of the donut. Retention is achieved by sliding vortices on small particles along zero electric field lines.

An optical bottleneck differs from an optical vortex in that it has a zero electric field only at the focus and non-zero electric fields in all other directions around the focus, i.e. at the ends of the vortex. Optical bottlenecks can be used to trap atoms and nanoclusters that are too small or too absorbing to be captured by optical vortex or optical tweezers. (J.Arlt & MJpadgett. "Gneration of a beam with a dark focus surrounded by regions of higher intensity: The optical bottle beam," Opt. Lett.25, 191-193, 2000.)

The optical rotator is a recently documented optical tool that provides a pattern of helical arms that capture a target. Changing the mode causes the captured target to rotate. (L.paterson, M.P.MacDonald, RArlt, RSibbett, P.E.Bryant, and K.Dholakia, "Controlled rotation of optically trapped microscopic particles," Science 292, 912-914, 2001.) Such tools can be used to control non-spherical particles And drive MEMs devices or nanomechanical devices.

Optical cages (Neal US Pat. No. 5,939,716) broadly speaking, are macroscopically the same family as optical vortices. The photocage forms a time-averaged ring of optical tweezers to surround particles that are too large or reflective to be trapped, with a lower dielectric constant than the surrounding medium. However, unlike eddies, non-zero electric field regions are created. Optical vortices, while similar in use to optical tweezers, operate on the opposite principle.

A need exists for an analytical method and system in which the interaction of a probe with a target can be assessed without immobilizing the probe on a substrate. There also exists a need for methods and systems for forming configurable (and reconfigurable) probe arrays that maintain probe identity throughout an assay regardless of probe location. The present invention fulfills these and other needs, and provides further related advantages.

Summary of the invention

The present invention provides a novel and improved method and system for constructing, configuring and using three-dimensional probe arrays.

Create light traps in a container. Optical traps are created by directing a beam of light, such as a laser beam, at an optical element that changes the phases of the beams by patterning them to produce sub-beams. This sub-beam is in turn focused by a lens and creates the gradient conditions necessary for optical trapping. Then, each probe with known properties is added to the container. Probes are selected for a given assay, and each probe is then selected by its inclusion in an optical trap.

The number and quality of probes forming an array can be easily reconfigured by using optical traps to add, remove, or replace probes. In an array, the arrangement of the probes is also dynamic relative to each other, since the spatial relationship of the probes to each other can be changed while maintaining the identity of the probes selected to make up the array. Thus, the array and each of its probes are also moveable and positionable in three dimensions as a whole or individually within the container.

When a probe remains contained in an optical trap, regardless of whether it has been relocated in the container or not, and regardless of any change in the "order" of its spatial position in the array, it can be determined by knowing the identity of the optical trap containing the probe. Maintain the identity of the probe. In addition, an optical trap can pass a probe to another optical trap, and so on, while tracking the probe's optical trap custody chain to maintain the identity of the probe contained in the optical trap.

Other features and advantages of the present invention will be set forth in part in the following specification and in the accompanying drawings, which describe and show preferred embodiments of the invention, and in part, after careful study of the following detailed description taken in conjunction with the accompanying drawings , which will be obvious to those skilled in the art, or can also be learned through practice of the present invention. The advantages of the invention may be realized and obtained by means of the instrumentalities and combinations particularly pointed out in the appended claims.

Description of drawings

Figure 1 shows a partial cross-sectional side view of a system for forming a configurable probe array.

Figure 2 shows a free probe contained within an optical trap.

Figure 3 shows a schematic diagram of the system used to form probe arrays.

Figure 4 shows the beam changing element with multiple static regions.

Figure 5A shows the first effective movement of the probe.

Figure 5B shows the second effective movement of the probe.

Figure 6A shows a compositional view of a miniature system for forming optical traps.

Figure 6B shows the inverted microscope of Figure 6A equipped with the miniature system.

Detailed description of the preferred embodiment

To illustrate the principles and operation of the invention, several specific embodiments of the invention are described in some detail below. However, various changes are possible, and the scope of the present invention is not limited by the exemplary embodiments described below. For example, while specific reference is made to biological systems and assays for gene sequences and DNA hybridization, it is understood that the methods and systems are equally useful in the fields described below, such as optical circuit fabrication and testing, nanocomposite construction and testing, optoelectronic devices Fabrication, electronic component testing, assembly and testing of holographic data storage matrices, chemical analysis, genomic analysis, proteomic analysis, facilitation of combinatorial chemistry, facilitation of self-assembly of colloids, and detection of non-biological materials.

Certain technical terms are used in the following description for convenience and by way of reference but not limitation. A short definition is provided below:

A. "Sub-beam" means a sub-beam of light or other energy source produced by directing a beam of light or other energy source, such as a beam produced by the collimated output of a laser or light emitting diode, through a medium, wherein the medium converts it diffracted into two or more sub-beams. An example of a sub-beam would be a higher order laser beam that diffracts off the grating.

B. "Phase profile" means the phase of light or other energy source in a cross-section of a beam or sub-beam.

C. "Phase patterning" means a patterned phase shift imparted to a beam or beamlet that alters the phase profile of the beam or beamlet, including, but not limited to, diffraction, phase modulation, patterning Forming, splitting, converging, dispersing, shaping, and otherwise manipulating beams or sub-beams.

D. "Probe" refers to a biological or other chemical material that selectively binds to or reacts with a target. Probes include, but are not limited to, oligonucleotides, polynucleotides, chemical compounds, proteins, peptides, lipids, polysaccharides, ligands, cells, antibodies, antigens, organelles, lipids, blastomeres, cell aggregates , microorganisms, cDNA, RNA and more.

E. "Target" means a biological or other chemical material whose presence or absence in a sample is detected by binding or reaction of the target to a probe. For example, the existence of a target formed by genetic material is through the reaction of the genetic material of the target and the genetic material of the probe (which has a specific characteristic necessary for hybridization, i.e. complementary structure), such as a hybridization reaction, And was detected. Target materials also include, but are not limited to, oligonucleotides, polynucleotides, chemical compounds, proteins, lipids, polysaccharides, ligands, cells, antibodies, antigens, organelles, lipids, blastomeres, cell aggregates, microorganisms, Peptides, cDNA, RNA and more.

As shown in FIG. 1, the probes 500-504 may be bound or reacted to any suitable substrate by any suitable bonding process or protocol. An important property of a suitable substrate is that it is a material that can be contained or manipulated by light traps. Typical dielectric substrates include beads, irregular small particles, or other regular small particles. Materials comprising suitable substrates include, but are not limited to, control pore glass, ceramics, silica, titanium dioxide, latex, plastics such as polystyrene, methylstyrene, polymethylmethacrylate Esters, paramagnetic materials, thoriosol, graphite, polytetrafluoroethylene, cross-linked dextran such as agarose, cellulose, nylon, cross-linked micelles, liposomes, and vesicles.

As shown in another alternative embodiment shown in FIG. 2, the method of the present invention also includes using one or more optical traps 1005 (one shown) to contain one or more probes that are not bound to the substrate. 505 (display one). It should be understood that a configurable array may contain only bonded probes, non-bonded probes, or a combination of bonded and non-bonded probes. What mixture of bonded and non-bonded probes is chosen, if any, may be influenced in part by the physical characteristics of the probes. In particular, the properties of certain probes, such as skin cells, can be altered so that they do not adhere to the substrate. Conversely, the effects of other probes, such as proteins, can be better exploited by removing the substrate to retain the third structure of the probe/protein.

Figure 1 shows a configurable array 8 of substrate-bonded probes 500-504 for analysis of biological material. Probes are deployed in the subject cell 10 using movable optical traps 1000-1004 constructed from focused sub-beams 2000-2004. The main cell 10 is a vessel constructed of a substantially transparent material that allows the sub-beams to pass through without affecting the formation of light traps.

Figure 3 shows a schematic diagram of a system, generally designated 20, for generating and changing the position of a configurable probe array. By propagating collimated light, preferably laser beam 100 generated by laser 102, into region A' on beam splitter 30, movable optical traps 1000-1004 are created in container 10 (FIG. 1). One of the beams, beam 31, comes from laser 102 and is redirected to proceed from region A' on beam splitter 30 to region A on phase patterning optical element 22. Each sub-beam generated by the phase-patterning optical element 22 then passes through a region B located at the rear aperture 28 of the focusing lens 12 . The sub-beams are converged by the focusing lens 12 . The resulting focused beamlets form optical traps 1000-1004 by creating the gradient conditions necessary to contain and control the probe in three dimensions. For clarity, only five sets of probes, sub-beams and optical traps are shown in Figure 1, but it should be understood that, depending on the nature, scope and other parameters of the assay and the capabilities of the system producing the optical traps, more or less less quantity.

Any suitable laser may be used as the energy source for laser beam 100 . Available lasers include solid state lasers, diode pump lasers, gas lasers, dye lasers, alexanderite lasers, free electron lasers, VCSEL lasers, diode lasers, Ti-sapphire lasers, doped YAG lasers, doped YLF lasers, diode-pumped YAG lasers, and flashlamp-pumped YAG lasers. Diode pumped Nd:YAG lasers operating between 10 mW and 5W are preferred. Preferred wavelengths of the laser beam 100 for forming arrays for studying biological materials include: infrared, near infrared, visible red (visible red), green, and visible blue (visible blue) wavelengths, with wavelengths from about 400 nm to about 1060 nm. wavelength is most preferred.

The beam splitter 30 is composed of a dichroic mirror, a photonic band gap mirror, an omnidirectional mirror, or other similar devices. The beam splitter 30 selectively reflects wavelengths of light used to form optical traps, and transmits other wavelengths. A portion of the light reflected from the beam splitter region A' then passes through region A of the encoded phase patterning optical elements 22 disposed substantially in the plane of the focusing lens 12 with the rear aperture 28 The plane 24 of the mating.

When the laser beam 100 is directed through the phase-patterning optical element 22, the phase-patterning optical element produces a plurality of sub-beams with altered phase profiles. Depending on the number and type of optical traps desired, such alterations can include diffraction, wavefront shaping, phase shifting, steering, dispersion and convergence. Based on the selected phase profile, phase-patterned optical elements can be used to generate optical traps in the following forms: optical tweezers, optical vortices, optical bottlenecks, optical rotators, optical cages, and combinations of two or more of these forms .

In those embodiments where the phase profile of the sub-beams is less intense at the periphery and more intense in regions inward from the periphery, less than about 15% of the back aperture 28 is overfilled, compared to a back aperture 28 that is not overfilled, with to form optical traps with greater intensity at the periphery.

Suitable phase-patterning optical elements can be characterized as transmissive or dioptric reflective, depending on how they orient the focused light beam or other energy beam. A catadioptric optical element transmits a light beam or other energy beam, while a catadioptric optical element reflects a light beam or other energy beam.

Phase patterning optical elements can also be classified as having static surfaces or having dynamic surfaces. Examples of suitable static phase patterning optical elements include those having one or more fixed surface areas, such as gratings, including scattering, reflection, and transmission gratings, holograms, including polychromatic holograms, templates, light Shaping hologram filters, polychromatic holograms, lenses, mirrors, prisms, waveplates and more. A static transmissive phase-patterning optical element 40 , as shown in FIG. 4 , features a fixed surface 41 . However, in some embodiments the phase patterning optic itself is movable, allowing selection of more than one fixed surface region 42-46 by moving the phase patterning optic relative to the laser beam to select the appropriate region. The static phase patterning optical element may be fixed on a spindle 47 and rotated about a controllable motor (not shown). The static phase patterning optical element in the embodiment shown in Figure 4 has a fixed surface 41 and discrete regions 42-46. In other embodiments of the static phase patterning optical element, whether transmissive or reflective, the fixed surface 41 has a non-uniform surface comprising substantially continuously varying regions, or a combination of discrete and substantially continuously varying regions.

Examples of suitable static phase-patterned optical elements whose function is time-dependent include: computer-generated diffraction patterns, phase-shifting materials, liquid crystal phase-shifting arrays, micromirror arrays, including pistonic micromirror arrays, spatial light modulators, electro-optic deflection devices, acousto-optic modulators, deformable mirrors, reflective MEMS arrays, and more. With dynamic phase-patterning optical elements, media containing phase-patterning optical elements encode holograms that can be altered to impart a patterned phase shift to the focused beam, which results in a corresponding change in the phase profile of the focused beam, e.g. Diffraction, or convergence. Additionally, the medium can be altered to produce changes in the position of the optical traps. The medium can be changed to move each light trap independently, which is an advantage of dynamically phase-patterned optics.

Preferred dynamic optical elements include phase-only spatial light modulators such as the "PAL-SLM Series X7665" manufactured by Hamamatsu, Japan, or the "SLM512N15' and SLM512SA7" manufactured by Boulder Nonlinear Systems of Layafette, Colorado. These phase patterning optical elements are computer controlled to generate beamlets 2000-2004 (FIG. 1) through holograms encoded in a medium that can be altered to generate beamlets and select the form of the beamlets.

In some embodiments, the form and/or position of the optical traps used to form the array are changed, configured and reconfigured accordingly. The form can be changed from its original form to the following: optical tweezers, optical vortex, optical bottleneck, optical rotator or optical cage. Light traps can move in two or three dimensions.

Phase patterning optics can also be used to impart a specific topological mode to the laser, for example, by converting Gaussian modes to Gaussian-Laguerre modes. Thus, one sub-beam can be formed into a Gaussian-Laguerre mode, while the other sub-beam can be formed into a Gaussian mode.

The probe is arranged in the container 10 . The container 10 is a main cell composed of a substantially transparent material, which allows the sub-beams to pass through without affecting the formation of optical traps. In those embodiments, where the substrate is labeled with a wavelength-specific dye, the principal cells should be transparent to that particular wavelength. Furthermore, the host cell should consist of 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 host cell and must not be altered or damaged by the material.

Probes with specific properties necessary to bind and/or react with a target of interest are selected for addition to the containers and inclusion into the configurable array. In some embodiments, where the probes are bonded to the substrate, the substrate is labeled (eg, a wavelength-specific dye) to facilitate probe selection. In a preferred embodiment, all substrates to which probes having the same binding or reactivity properties are bound are labeled with the same type of label. When the substrate is marked with a wavelength-specific label, the selection of probes 500-504 can be accomplished by adding probes to the container 10 that are bonded to the labeled substrate. Then, as shown in Figure 3, spectroscopic measurements of the labeled substrates of the probes can be used to select (or not select) probes for inclusion in the array. In some embodiments (FIG. 2) the probes may not be bound to the substrate and may also be labeled.

In embodiments where unlabeled probes are selected to form all or part of the array, the probes may be added to container 10 sequentially. In such cases, the identity of the probes can be known by the order of loading or the identity of the probes can be known by the time of addition of the probes. Alternatively, probes having different binding properties or reactivity properties from each other may be segregated to different predetermined locations according to the different properties. Probes are then selected based on their position in the container.

As seen in Figure 3, the spectroscopy of a sample of biological material can be accomplished with an imaging illumination source 39 suitable for both spectroscopy and polarized lightback scattering, the former being used to assess chemical identity, The latter are suitable for measuring the dimensions of internal structures, such as atomic nucleus dimensions. In some embodiments, using such spectroscopic methods, cells are interrogated and probe arrays are created from selected interrogated cells. For example, computer 38 can be used to analyze spectroscopic data and to identify suspected cancerous, precancerous and/or noncancerous cell types. A computer can then use the information to guide the light trap to contain the selected cell type. The contained cells can then be used as probes in assays based on the reaction or binding of the contained cells to targets such as other cells, antibodies, antigens, and other biological materials, or pharmaceuticals and other chemicals. Those skilled in the art will recognize that, depending on parameters specific to cancer cells, the methodology for interrogating and concentrating cells may be altered for interrogating and/or isolating blastomeres, cells, or other material without departing from this disclosure.

the scope of the invention.

In other embodiments, labeled or unlabeled probes, eg, unlabeled probes with different binding or reactivity properties, may be placed in a series of daughter cells 16 disposed in container 10 . In Figure 1, only one daughter cell is shown for clarity. However, it should be understood that a plurality of such daughter cells may be provided. In some embodiments, the boundaries of the daughter cells are formed by light traps. A number of optical traps placed in the correct orientation create an optical sub-cell that can perform the same function as the physical sub-cell 16 .

Arrangement of the probes in daughter cells 16 is by any suitable means, including trapping with light, moving through fluidic channels, through microcapillaries, or by other equivalent mechanisms. In each daughter cell, one or more probes with the same binding or reactivity properties are placed. Then, the probes included in the array are selected based on the daughter cells containing the probes.

The selected probes 500-504 are then captured using the optical traps 1000-1004 by including the probes in the optical traps 1000-1004. A set of such contained probes is thus configured to form an array.

The inventive method and system itself provides a semi-automatic or automatic process for tracking the movement and content of each light trap. This movement can be monitored by camera, spectral, or optical data streams, which provide computer-controlled probe selection, and generation of optical trap information for adjusting the type of probe contained in the optical trap, and formation of the array. Components of the probe. In other embodiments, the movement is tracked according to a predetermined movement of each optical trap induced by encoding the phase-patterned optical element. Additionally, in some embodiments, a computer is used to maintain a record of each probe contained in each optical trap.

Returning to the beam splitter 30, the beam splitter 30 also provides a light beam 32 from an imaging illumination source 39 which passes through the main cell 10 to form one or more beams derived from the orientation and position of the probe contained in the optical trap. The optical data stream corresponding to the sub-beam.

The optical data stream may then be observed, converted to a video signal, monitored, or analyzed by visual inspection 34a of an operator 36, using a spectroscopic device 34b, and/or video monitoring 34c. Optical data stream 32 may also be processed by a photodetector monitoring intensity or any suitable device to convert the optical data stream to a digital data stream for use by computer 38 .

To construct the array, the operator 36 and/or computer 38 adjusts the hologram encoded by the phase-patterning optical element 22 to guide the movement of each optical trap to obtain the selected probe and capture it. A plurality of phototrap-forming array components with contained probes, the array can be reconfigured according to the needs of the user as to the composition or location of the probes. Using the optical data stream, the location of one or more captured probes can be identified and their location can be monitored. Based on such information, the surface of the phase-patterning optical element can be altered, in some embodiments independently, to alter the form of one or more optical traps containing the probe.

Additionally, the position of one or more captured probes in the array can be tracked by monitoring the position of the optical trap containing it. Then, using such information, any given probe in the array can be independently relocalized in the principal cell by altering the surface of the phase-patterning optical element, and through the probe-containing optical trap, each probe's The identity remains known regardless of where the phototrap positions the probe.

In a preferred embodiment, computer 38 controls the movement of the optical trap both before and after capture of the probe. In other embodiments, the optical data stream is first converted to a video signal, which is then used to generate an image corresponding to the array, and an operator then observes the image to control the movement of at least one optical trap based on the image.

Referring to Figures 1 and 3, for analysis, a first batch of targets T1-T5 is added to the main cell 10 through the inlet 14, which also contains a fluid medium 3000. Arrays of probes 500-504 are suspended in medium 3000 by optical traps 1000-1004 to their containment. To increase the chance of reactivity with target T1-T5, the probe can be moved around the principal cell corresponding to the movement of the light trap.

For example, in one embodiment, probes 500-504 are rotated through medium 3000 containing targets T1-T5. By containing the probe optically, as opposed to physically, and moving the probe within the host cell 10, the chances of the probe interacting with each target are increased, thus increasing the speed and efficiency of the assay.

An array of moving probes 500-502 is shown in Figures 5A and 5B by sequentially generating sets of optical traps. In the embodiment shown in FIG. 5A, a simple linear movement of the probe array is shown, which is configured along the line P1, where P1 represents the first set of predetermined positions. Movement is accomplished by transferring the probes from the first set of optical traps to the second, third, and then fourth set. With additional reference to FIG. 4 , the first set of optical traps is created by directing a laser beam on the first region 42 of the phase patterning optical element 40 . When the sub-beams emitted by the first region 42 pass through the focusing lens, they form a first set of optical traps at the first position P1 containing the probes 500-503.

To move the probes 500-502 from the first position P1 to the second position P2, the static phase-patterning optical element 40 is rotated about the axis 47 to align the laser beam with the second region 43, at the corresponding second set of predetermined A second set of optical traps is created at position P2. Probes can be transferred from the first set of optical traps to the second set of optical traps by constructing the second set of optical traps in appropriate proximity to the first position P1. By rotating the phase-patterning optical element to align the appropriate regions 42-46 corresponding to the desired positions P1-P5, the probe can be sequentially passed from the second set of predetermined positions P2 to the third set of predetermined positions P3, from the third set of predetermined positions P3 to the third set of predetermined positions P3. The group of predetermined positions P3 to the fourth group of predetermined positions P4, and from the fourth group of predetermined positions P4 to the fifth group of predetermined positions P5. The time interval between the termination of one set of optical traps and the generation of the next should be of some duration to ensure that the probe passes to the next set of optical traps before drifting away.

This movement of the probe can be used to troll the probe through the medium, thereby increasing the chances of the target in the medium interacting with the probe. This type of simple movement can also be used to move a probe from a daughter cell 16 ( FIG. 1 ) to another area of the main cell 10 , or to sequester a probe into a daughter cell 16 .

In the example shown in FIG. 5B , a staggered movement of the probes in close proximity from wide to narrow is shown. The staggered movement of the probes occurs in a manner similar to that described with reference to Figure 5A. However, now the first region 42 generates optical traps with the two probes 500 and 502 arranged along the line P1, while the third probe 501 is arranged at P2, between the above two probes, but with the line P1 Spaced out. As the probes pass from the first set of optical traps to the second and move to the second and subsequent positions, the staggered arrangement of the probes allows the probes to be densely packed without having to be placed too close to two probes at the same time A set of traps that would otherwise result in the probe being included in the wrong light trap.

Once the target has interacted with the probe, the target can be studied using spectroscopic methods. The spectra of those probes with positive results (ie those reacted or bound to the target) can be obtained by using image illumination 39, eg this is suitable for inelastic spectroscopy or polarized light backscattering. Computer 38 can analyze the spectroscopic data to identify desired targets, and direct the phase patterning element to isolate those desired targets. Those skilled in the art will recognize that the methodology for isolating targets based on spectral data may be altered to identify and/or isolate targets based on other information available from the target and/or optical data stream without departing from this disclosure. the scope of the invention.

When the analysis is complete, via the computer 38 and/or the operator 36, a choice is made as to which probes to discard and which probes to collect. The reconfigurable nature of the array allows the selective movement of a given optical trap and contained probe. In some cases, the media 3000 and non-adhered targets are expelled or washed from the host cell 10 via the outlet 18, and the analysis is then completed. In other cases, at least some of the probes still contained in the optical traps are reused for further analysis, along with additional targets. Depending on the parameters of the analysis, this technique can be used in the case of probes that are determined to be positive or negative. In still other cases, as the array of probes is reconfigured with respect to the number and identity of probes forming the array, optical traps can be used to exclude non-adhesive probes and obtain additional probes for further experiments.

In some embodiments, it is not necessary to generate sub-beams from each region of the static beam modifying optical element 40, or to move the beam modifying optical element 40 in a prescribed direction. Alternatively, changing the order of the regions changes the position of the set of light traps.

Figure 6A shows a perspective view of a compact system, generally designated 50, for forming optical traps. Phase patterning optical element 51 is a dynamic optical element, having a reflective, dynamic surface, which is also a phase-only spatial light modulator such as the "PAL-SLM Series X7665" manufactured by Hamamatsu, Japan, or by Nonlinear Systems of Boulder, Colorado. "SLM512N15' and SLM512SA7" manufactured by Lafayette. These dynamic optical elements feature coded reflective surfaces where a computer can control the holograms formed within them.

Figure 6A shows a compact system for forming optical traps with an optical element 51 aligned or attached to a housing 52 through which a first optical channel 53a is provided. One end 53b of the first optical channel is adjacent to the optical element 51, and the other end 53c of the first optical channel interacts with and communicates with the second optical channel 53d perpendicular thereto. The second optical channel is formed in the base 54a of the microscope lens mounted on a turret or "nosepiece" 54b. The nosepiece 54b is adapted to fit into a Nixon TE 200 series microscope (not shown). The second optical channel communicates with a third optical channel 55a which is also orthogonal to the second optical channel. The third optical channel 55a traverses from the top surface of the nosepiece 54b through the nosepiece 54a and is parallel to the objective focusing lens 56 . The focusing lens has a top and a bottom forming a rear aperture 57 . Inserted for the third optical channel between the second optical channel and the rear aperture 57 of the focusing lens is a dichroic mirror beam splitter 58 . Other components used to form the optical trap 50 in a compact system include a first mirror M1 that reflects the sub-beams emitted from the phase-patterned optical element through the first optical channel in which the first set of transmission The optical device TO1, which is collimated to receive the sub-beam reflected by the first mirror M1, the second group of transfer optical device TO2 arranged in the first optical channel, which is collimated to receive the sub-beam passed through the first group of transfer lenses TO1 The sub-beams, and the second mirror M2 located at the intersection of the first and second optical channels, are collimated to reflect the sub-beams passing through the second set of transfer optics TO2 and the third optical channel 55a.

To create optical traps, a laser beam (not shown) is directed through optical fiber 150 out of collimator end 151 and reflected off dynamic surface 59 of optical element 51 . The light beam (not shown) output from the collimator end 151 of the optical fiber 150 is diffracted by the dynamic surface 59 of the optical element 51 into a plurality of sub-beams (not shown). The numbering type and direction of each sub-beam can be controlled and changed by altering the hologram encoded in the dynamic surface medium 59 . The sub-beams then reflect off the first mirror M1, pass through the first set of transfer optics TO1, pass through the second set of transfer optics TO2 along the first optical channel 53a to the second mirror M2; are then directed on the beam splitter 58 until The rear aperture 57 of the objective lens 56 is converged by the objective lens 56 to create the optical gradient conditions necessary to form the optical traps. For imaging, the part of the light split by the dichroic mirror 58 passes through the lower part of the third optical channel 55b to form an optical data stream (not shown).

In those embodiments in which the phase profile of the sub-beams is less intense at the periphery and more intense in regions inward from the periphery, less than about 15% of the rear aperture 57 is overfilled compared to a rear aperture 57 that is not overfilled. , for the formation of light traps with greater intensity around them.

Figure 6B shows a perspective view of a Nixon TE 200 series microscope equipped with a small system, generally designated 60, for forming optical traps 50. The nosepiece 54 with the housing 52 attached thereto is mounted directly in the microscope by a base supporting the nosepieces 54a and 54b. The housing and its contents and associated optics 51 are secured to nosepieces 54a and 54b, requiring little or no changes or modifications to the rest of the microscope. For imaging, an illumination source may be provided above the objective lens 56 .

The first and second sets of transfer optics TO1 and TO2 are shown each containing two lens elements. Lenses can be convex or concave. Different and varying types and numbers of lenses, such as symmetrical air spaced singlets, symmetrical air spaced doublets and/or additional lenses or lens groups, may be selected to achieve the image Transfer from the first mirror M1 to the second mirror M2. In some embodiments, the first and second sets of transfer optics are symmetrical air-gap doublets that are spaced apart and combined as a telephoto lens.

Since certain changes can be made in the above system devices and methods without departing from the scope of the present invention, all the content contained in the above description, as shown in the drawings and specification, should be interpreted as illustrative rather than meaning of restriction.

Claims (157)

1, the method for a kind of configuration and tracking probe array comprises:

In container, produce at least two movably light trappings;

At least two probes are provided in container;

Select at least two probes to be used for the inclusion of the probe array that light trapping inside comprised;

With catching each selected probe one of in the light trapping, be contained in the probe array of light trapping inside with configuration packet; And,

Comprise the position of the light trapping of probe, the position of at least one captive probe in the tracking array by monitoring.

2, the method in the claim 1 further comprises by moving the light trapping that comprises tracked probe and changes the position of at least one tracked probe.

3, the method in the claim 1, wherein light trapping is by two or more light tweezers, the optics whirlpool, the optics bottleneck, optical rotator or light cage constitute.

4, the method in the claim 2, wherein each light trapping can move independently.

5, the method in the claim 2, wherein each light trapping moves by computer control.

6, the method in the claim 4, wherein each light trapping moves by computer control.

7, the method in the claim 4, wherein at least one probe combines with the substrate that indicates the wavelength specific markers, and this at least one probe is by utilizing this mark of spectral measurement and utilizing spectral measurement to select this at least one bonded probe.

8, the method in the claim 4, wherein at least two probes have different combinations or response characteristic each other, and by selecting at least one probe, by traveling probe preposition and use the position of segregate probe to select this probe in the container by isolating probe according to the different combination of probe or response characteristic.

9, the method in the claim 8, wherein preposition is the daughter cell of physics.

10, the method in the claim 8, wherein preposition is the optical daughter cell.

11, the method in the claim 1 further comprises, introduces at least one target in the container, and determines that each captive probe and each target are with or without reaction therein.

12, the method in the claim 11, wherein captive probe is a biomaterial.

13, the method in the claim 11, wherein captive probe is a chemical compound.

14, the method in the claim 12, it hits is biomaterial.

15, the method in the claim 12, it hits is chemical compound.

16, the method in the claim 13, it hits is biomaterial.

17, the method in the claim 13, it hits is chemical compound.

18, the method in the claim 12, wherein captive probe is an oligonucleotide, polynucleotide, protein, polysaccharide, ligand, cell, antibody, antigen, honeycomb cell organelle, fat, blastomere, cell aggregation, microorganism, peptide, cDNA, RNA or their combination.

19, the method in the claim 14, it hits is oligonucleotide, polynucleotide, protein, polysaccharide, ligand, cell, antibody, antigen, honeycomb cell organelle, fat, blastomere, cell aggregation, microorganism, peptide, cDNA, RNA or their combination.

20, the method in the claim 16, it hits and is selected from by oligonucleotide, polynucleotide, protein, polysaccharide, ligand, cell, antibody, antigen, honeycomb cell organelle, fat, blastomere, cell aggregation, microorganism, peptide, cDNA, in the group that RNA or they constitute one or more.

21, the method for claim 1 further comprises, probe all combines with substrate.

22, the method for claim 1 comprises that further probe all directly caught by light trapping.

23, the method for claim 1 comprises that further at least some probes combine with substrate and at least some probes do not combine with substrate.

24, the method for claim 21 further comprises, by moving the position of at least two tracked probes in the light trapping change array that comprises probe.

25, the method for claim 1 further comprises the optical data stream that produces with the corresponding data of identity and position of at least one light trapping.

26, the method for claim 24, wherein each light trapping can move independently.

27, the method for claim 24, wherein each light trapping mobile be subjected to computer-controlled.

28, the method for claim 25 further comprises with computer receiving optical data stream.

29, the method for claim 28 further comprises and uses the Computer Analysis optical data stream.

30, the method for claim 29, wherein computer moving according at least one light trapping of analysis and guidance of optical data stream.

31, the method for claim 25 further comprises optical data stream is converted to vision signal.

32, the method for claim 31 further comprises and uses the computer receiving video signals.

33, the method for claim 32 further comprises and uses the Computer Analysis vision signal.

34, the method for claim 33 comprises that further the analysis and utilization computer according to vision signal instructs moving of one or more light trappings.

35, the method for claim 31, wherein vision signal is used to produce image.

36, the method for claim 35 comprises that further the operator observes image, and instructs moving of one or more light trappings based on image observation.

37, the method for claim 25, wherein data are spectroscopic datas.

38, the method for claim 37 comprises that further the analysis and utilization computer based on spectroscopic data instructs moving of one or more light trappings.

39, the method for claim 24, wherein light trapping is by two or more light tweezers, the optics whirlpool, the optics bottleneck, optical rotator or light cage form.

40, the method for claim 26, wherein each light trapping moves by computer control.

41, the method for claim 24 is wherein by spectral measurement mark and utilize results of spectral measurements to select at least one probe.

42, the method for claim 24, wherein at least two probes have combination or the response characteristic that is different from each other, and select at least one probe by isolating this probe, by traveling probe preposition and use the position of segregate probe to select this probe in the container according to the different combination of probe or response characteristic.

43, the method in the claim 42, wherein preposition is the daughter cell of physics.

44, the method in the claim 42, wherein preposition is the optical daughter cell.

45, the method in the claim 169, wherein captive probe is a biomaterial.

46, the method in the claim 169, wherein captive probe is a chemical compound.

47, the method in the claim 46, it hits is biomaterial.

48, the method in the claim 46, it hits is chemical compound.

49, the method in the claim 45, it hits is biomaterial.

50, the method in the claim 45, it hits is chemical compound.

51, the method in the claim 45, wherein captive probe is an oligonucleotide, polynucleotide, protein, polysaccharide, ligand, cell, antibody, antigen, honeycomb cell organelle, fat, blastomere, cell aggregation, microorganism, peptide, cDNA, RNA or its combination.

52, the method in the claim 47, it hits is oligonucleotide, polynucleotide, protein, polysaccharide, ligand, cell, antibody, antigen, honeycomb cell organelle, fat, blastomere, cell aggregation, microorganism, peptide, cDNA, RNA or its combination.

53, the method in the claim 49, it hits and is selected from by oligonucleotide, polynucleotide, protein, polysaccharide, ligand, cell, antibody, antigen, honeycomb cell organelle, fat, blastomere, cell aggregation, microorganism, peptide, cDNA, in the group that RNA or its constitute one or more.

54, the method for claim 24, its middle probe all combines with substrate.

55, the method for claim 24, its middle probe does not all combine with substrate.

56, the method for claim 24, wherein at least some probes combine with substrate and at least some probes do not combine with substrate.

57, a kind of method of analysis of biological material comprises:

Produce at least two movably light trappings at internal tank;

Fluid medium is provided in container;

At least two probes that are used for biomaterial are provided in fluid medium;

Select at least two probes to be used for the inclusion of array;

Follow the tracks of each selecteed probe with one of light trapping;

At least one comprises the target of biomaterial in the introducing container; With,

Determine each captive probe and each target response or not reaction.

58, the method for claim 57 further comprises the position that comprises the light trapping of probe by monitoring, follows the tracks of the position of at least one captive probe.

59, the method for claim 57, wherein captive probe comprises biomaterial.

60, the method for claim 57, wherein captive probe comprises chemical compound.

61, the method for claim 59, wherein captive probe is an oligonucleotide, polynucleotide, protein, polysaccharide, ligand, cell, antibody, antigen, honeycomb cell organelle, fat, blastomere, cell aggregation, microorganism, peptide, cDNA, RNA or its combination.

62, the method for claim 57, it hits is oligonucleotide, polynucleotide, protein, polysaccharide, ligand, cell, antibody, antigen, honeycomb cell organelle, fat, blastomere, cell aggregation, microorganism, peptide, cDNA, RNA or its combination.

63, the method for claim 57 further comprises, the optical data stream of the corresponding data of identity and position of generation and at least one light trapping.

64, the method for claim 63 further comprises, by moving the position of at least one captive probe in the light trapping change array that comprises probe.

65, the method for claim 64, wherein each light trapping can move independently.

66, the method for claim 64, wherein each light trapping mobile be subjected to computer-controlled.

67, the method for claim 63 further comprises with computer receiving optical data stream.

68, the method for claim 67 further comprises and uses the Computer Analysis optical data stream.

69, the method for claim 68 further comprises and utilizes computer based moving in the one or more light trappings of analysis and guidance of optical data stream.

70, the method for claim 63 further comprises optical data stream is converted to vision signal.

71, the method for claim 70 further comprises and uses the computer receiving video signals.

72, the method for claim 71 further comprises and uses the Computer Analysis vision signal.

73, the method for claim 72 further comprises and utilizes computer based moving in the one or more light trappings of analysis and guidance of vision signal.

74, the method for claim 70, wherein vision signal is used to produce image.

75, the method for claim 74 comprises that further the operator observes image, and instructs moving of one or more light trappings based on image observation.

76, the method for claim 63, wherein data are spectroscopic datas.

77, the method for claim 76 further comprises and utilizes computer based moving in the one or more light trappings of analysis and guidance of spectroscopic data.

78, the method for claim 63, wherein light trapping is by two or more light tweezers, the optics whirlpool, the optics bottleneck, optical rotator or light cage form.

79, the method for claim 63, wherein at least one probe combines with substrate.

80, the method for claim 63, wherein at least one probe does not combine with substrate.

81, the method for claim 79, what wherein all had identical combination or a response characteristic indicates identical mark with substrate bonded probe.

82, the method for claim 81, wherein at least one mark is the wavelength particular dye.

83, the method for claim 82, wherein for selecting at least one probe, the spectral response by measuring the wavelength particular dye and utilize results of spectral measurements to select at least one and substrate bonded probe.

84, the method for claim 63, wherein at least two probes have different combinations or response characteristic each other, and isolate this probe according to the different combination of probe or response characteristic and select at least one probe, by traveling probe preposition and utilize the position of segregate probe to select this probe in the container.

85, the method in the claim 63, wherein preposition is the daughter cell of physics.

86, the method in the claim 84, wherein preposition is the optical daughter cell.

87, a kind of method that disposes probe array comprises:

Produce at least two movably light trappings at internal tank;

Provide at least two probes at internal tank; With,

By selecting each probe to dispose the array of at least two probes with one of light trapping.

88, a kind of method that disposes and dispose probe array comprises:

A directed focused beam on the phase pattern optical element to form a plurality of beamlets of locating the phase pattern optical element;

Directed these a plurality of beamlets pass through condenser lens to transmit beamlet, and assemble the beamlet that sends from condenser lens, to produce movably light trapping in container on the condenser lens back aperture;

A large amount of probes are provided in container;

Select at least two probes to be used for being included in the content of the probe array in the light trapping;

Catch each chosen probe with one of light trapping, be contained in probe array in the light trapping with configuration packet; With

By moving the light trapping comprise probe, change the position that at least one is included in the interior probe of light trapping, be contained in probe array in the light trapping with configuration packet again.

89, the method for claim 90, wherein the phase pattern optical element has static surface.

90, the method for claim 91, wherein Jing Tai surface comprises two or more discrete zones.

91, the method for claim 90, wherein at least one position that is contained in the probe in the light trapping is changed by the zone of dispersion of the static surface that changes light beam and be directed to.

92, the method for claim 89, wherein static surface is basic continually varying.

93, the method for claim 89, wherein the position of at least one light trapping is by the zone of dispersion change of the static surface that changes light beam and be directed to.

94, the method for claim 89, wherein light beam change optical element is a grating, hologram, masterplate, polishing shape holographic filter, lens, speculum, prism, or wave plate.

95, the method for claim 90, wherein each zone of dispersion is a grating, hologram, masterplate, polishing shape holographic filter, lens, speculum, prism, or wave plate.

96, the method for claim 88, wherein the phase pattern optical element is dynamic.

97, the method for claim 96, wherein at least one position that is contained in the probe in the light trapping is changed by changing dynamic phasing medelling optical element.

98, the method for claim 97, wherein the form of at least one light trapping is changed by changing dynamic phasing medelling optical element.

99, the method for claim 97, wherein the light trapping of Gai Bianing is the light tweezer, optics whirlpool, optics bottleneck, optical rotator or light cage.

100, the method for claim 91, wherein the form of at least one light trapping changes by moving discrete static surface.

101, the method for claim 100, wherein reformed light trapping is the light tweezer, optics whirlpool, optics bottleneck, optical rotator or light cage.

102, the method for claim 97, wherein the change of dynamic phasing medelling optical element is the variation in the hologram of encoding in its surface.

103, a kind of system that is used to form and follows the tracks of the light trapping that comprises probe comprises:

Be used to produce the light source of focused beam;

The container of substantially transparent;

Be used for producing the image light source of light beam of the inclusion of illumination container;

Be used for directed beam splitter;

The phase pattern optical element, it is used to receive the focused beam from light source, and it is diffracted at least two beamlets, the phase pattern optical element has a surface and is used for the back aperture of directed each beamlet at condenser lens, and this surface is the phase section and/or the orientation that can change to change at least one beamlet;

Be used to assemble the condenser lens of each beamlet with the light trapping that is formed for comprising probe; With

Be used for receiving the light beam of the inclusion of illumination container, and follow the tracks of moving and the monitor of inclusion of at least one light trapping.

104, the system of claim 103 further comprises, includes the container of an inlet.

105, the system of claim 103 further comprises, includes the container of an outlet.

106, the method for claim 8, its middle probe are to utilize by light trapping, flow passage or microscopic capillary mobile and segregate.

107, the method for claim 42, its middle probe are to utilize by light trapping, flow passage or microscopic capillary mobile and segregate.

108, the method for claim 84, its middle probe are to utilize by light trapping, flow passage or microscopic capillary mobile and segregate.

109, the method for claim 63, it hits and is selected from by oligonucleotide, polynucleotide, protein, polysaccharide, ligand, cell, antibody, antigen, honeycomb cell organelle, fat, blastomere, cell aggregation, microorganism, peptide, in the group that cDNA and RNA constitute one or more.

110, the method for claim 9, its middle probe are to utilize by light trapping, flow passage or microscopic capillary mobile and segregate.

111, the system of claim 103, wherein the phase pattern optical element is dynamic, and it further comprises:

First computer is to control the diffraction that is caused by the phase pattern optical element; With,

Second computer is to keep being contained in the record of each probe in the light trapping.

112, the method for claim 2 wherein is based on the predetermined mobile of caused each light trapping of encoding phase medelling optical element, follows the tracks of moving of captive probe.

113, a kind of system that is used to form and follows the tracks of the light trapping that comprises the probe that is incorporated into target comprises:

With a plurality of probes of target bonded;

Be used to produce the light source of focused beam;

The container of substantially transparent;

The image light source is used for producing the light beam of the inclusion of illumination container;

Beam splitter is used for the light beam that orientation results from the inclusion of the focused beam of light source and the container that throws light on;

The phase pattern optical element, it is used to receive the light beam from light source, and it is diffracted at least two beamlets, the phase pattern optical element has a surface and is used for the back aperture of directed each beamlet at condenser lens, and this surface is changeable phase section and/or orientation to change at least one beamlet;

Condenser lens is used to assemble each beamlet and comprises light trapping with target bonded probe with formation; With

Monitor is used for receiving the light beam of the inclusion of illumination container, and follows the tracks of moving and inclusion of at least one light trapping.

114, the system of claim 113, its middle probe is a biomaterial.

115, the system of claim 113, its middle probe is a chemical compound.

116, the system in the claim 114, it hits is biomaterial.

117, the system in the claim 114, it hits is chemical compound.

118, the system in the claim 115, it hits is biomaterial.

119, the system in the claim 115, it hits is chemical compound.

120, the system in the claim 114, its middle probe is selected from by oligonucleotide, polynucleotide, protein, peptide, in the group that cDNA and RNA constitute one or more.

121, the system in the claim 116, it hits and is selected from by oligonucleotide, polynucleotide, protein, polysaccharide, ligand, cell, antibody, antigen, honeycomb cell organelle, fat, blastomere, cell aggregation, microorganism, peptide, in the group that cDNA and RNA constitute one or more.

122, the system in the claim 118, it hits and is selected from by oligonucleotide, polynucleotide, protein, polysaccharide, ligand, cell, antibody, antigen, honeycomb cell organelle, fat, blastomere, cell aggregation, microorganism, peptide, in the group that cDNA and RNA constitute one or more.

123, the method in the claim 2, wherein the mobile of at least one light trapping is selected from by the rotation in the fixed position, the rotation in the on-fixed position, in two dimension move and the group of the mobile formation in three-dimensional in one or more.

124, the method in the claim 2 further comprises by changing the surface of phase pattern optical element, moves the light trapping that comprises tracked probe.

125, the system in the claim 103, wherein the phase pattern optical element has static surface.

126, the system in the claim 125, wherein static surface comprises two or more zone of dispersions.

127, the system in the claim 126, wherein static surface is movably, so that focused beam is aimed at the chosen zone of static surface.

128, the method in the claim 2, wherein the phase pattern optical element has static surface, this static surface has two or more discrete zones, and the position of at least one light trapping is changed by the zone of dispersion of the static surface that changes light beam and be directed to.

129, the system in the claim 103, wherein the phase pattern optical element has basic continually varying static surface.

130, the system in the claim 127, wherein the phase pattern optical element is selected from and comprises grating, hologram, masterplate, light finishing hologram wave filter, lens, speculum, prism, or the group of wave plate.

131, the system in the claim 126, wherein each discrete zone is selected from and comprises grating, hologram, masterplate, light finishing hologram wave filter, lens, speculum, prism, or the group of wave plate.

132, the system in the claim 103, wherein the phase pattern optical element is dynamic.

133, the method in the claim 2, wherein the phase pattern dynamic element is dynamic, and changes the position that the phase pattern optical element changes at least one light trapping.

134, the method in the claim 4, wherein the phase pattern dynamic element is dynamic, and changes the position that the phase pattern optical element changes at least one light trapping.

135, the method in the claim 2, wherein the phase pattern dynamic element is dynamic, and to change the form that the phase pattern optical element changes at least one light trapping be the light tweezer, optics whirlpool, optics bottleneck, optical rotator or light cage.

136, the method in the claim 2, wherein the phase pattern optical element has static surface, and this static surface comprises two or more zone of dispersions, and the form of at least one light trapping is changed by mobile static surface.

137, the method in the claim 136, wherein the form of the light trapping of Gai Bianing is selected from by the light tweezer, optics whirlpool, optics bottleneck, the group that optical rotator or light cage constitute.

138, the system in the claim 132, wherein the phase pattern optical element is selected from least one in the group, this group comprises that computer generates diffraction pattern, phase shift material, liquid crystal phase shift array, micro mirror array, spatial light modulator, electro-optic deflector, acousto-optic modulator, distorting lens, reflection MEMS array.

139, the system in the claim 132 comprises that further a computer is with control dynamic phasing medelling optical element.

140, the system in the claim 103 comprises that further daughter cell is used to isolate the light trapping that at least one comprises probe in container.

141, the system in the claim 140, wherein daughter cell is the daughter cell of physics.

142, the system in the claim 150 further comprises a computer, forms the pattern optical element with change bit and changes the orientation of at least one beamlet and move corresponding light trapping to comprise probe.

143, the system in the claim 103, wherein light source is a laser, it is used to produce the focused beam with green spectral wavelength.

144, the system in the claim 103, wherein light source is a laser, it is used to produce the focused beam with visible light blue color spectrum wavelength.

System in 145 claims 103, wherein light source is a laser, it is used to produce the focused beam with visible light red spectrum wavelength.

146, the system in the claim 103, wherein light source produces and has the focused beam of about 400nm to the wavelength of about 1060nm scope.

147, the system in the claim 103, wherein light source is a laser beam.

148, the system in the claim 103 further comprises the computer that is used to receive optical data stream.

149, a kind of device that is used to form the light trapping array comprises:

Be used to produce the light source of light beam;

Condenser lens with top and bottom, back aperture is formed on described bottom;

The phase pattern optical element, it is used for the light beam of collectiong focusing, and it is diffracted at least two beamlets, and the phase pattern optical element has the back aperture that a surface is used for each beamlet is oriented in condenser lens;

First optical channel with first and second ends, first end communicates with the phase pattern optical element;

Second optical channel with first and second ends, first end intersects with second end of first optical channel;

The 3rd optical channel with first and second ends, first end communicates with second end of second optical channel;

First speculum, it is used to reflect the beamlet that sends from the phase pattern optical element and passes through first optical channel;

First group is transmitted Optical devices, and it is arranged in first optical channel, and it is collimated to receive the beamlet that first speculum is reflected;

Second group is transmitted Optical devices, and it is arranged in first optical channel, and it is collimated to receive the beamlet by first group of relay len;

Second speculum, it is positioned at the intersection point of first optical channel and second optical channel, and it is collimated with the beamlet of reflection by second group of transmission Optical devices and the 3rd optical channel; With

The 3rd speculum, its its be arranged within the 3rd optical channel, be used to reflect beamlet by the 3rd optical channel to the back aperture of condenser lens, thereby form the light trapping array.

150, the device of claim 149 further comprises, is used to produce the light source of illuminating bundle, and it is arranged on the top of proximity focusing lens.

151, the device of claim 150, wherein the 3rd speculum is a dichroic beam splitters, is used to guide the focused beam of light source generation and the light beam that light source produces.

152, the device of claim 149, wherein every group of transmission Optical devices are selected from single eyeglass in symmetrical clearance and symmetrical clearance doublet.

153, the device of claim 149, wherein every group is transmitted Optical devices and comprises the lens that are selected from the group that convex lens and concavees lens form.

154, the device of claim 149, wherein first and second groups of transmission Optical devices are symmetrical clearances, and keep at a certain distance away with combination as long shot.

155, the method for claim 25 further comprises and introduces at least one target of container, and determines that each captive probe and each target are with or without reaction therein.

156, the method consistent with claim 1 further comprises, by transmit probe from a light trapping to another light trapping, move at least one captive probe.

157, the method consistent with claim 1 further comprises, by transmit probe from first group of light trapping to second group of light trapping, move at least three captive probes.

Images