METHOD AND APPARATUS FOR SELECTIVELY TARGETING SPECIFIC CELLS WITHIN A CELL POPULATION

Abstract: This invention provides a method and apparatus for selectively identifying, and targeting with an energy beam, specific cells within a cell population, for the purpose of inducing a response in the targeted cells. Using the present invention, every detectable cell in a population can be identified and affected, without substantially affecting non-targeted cells within the mixture.

Description: METHOD AND APPARATUS FOR SELECTIVELY TARGETING SPECIFIC CELLS WITHIN A CELL POPULATION Background of the Invention Field of the Invention This invention relates to methods and apparatus for selectively targeting specific cells within a population of living cells. In particular, this invention relates to high-speed methods and apparatus for selectively identifying, and individually targeting with an energy beam, specific cells within a cell population to induce a response in the targeted cells. Description of the Related Art The use of cellular therapies is growing rapidly, and is therefore becoming an important therapeutic modality in the practice of medicine. Unlike other therapies, cellular therapies achieve a long-lasting, and often permanent benefit through the use of living cells. Hematopoietic stem cell (HSC) (e.g., bone marrow or mobilized peripheral blood) transplantation is one example of a practiced, insurance-reimbursed cellular therapy. Many other cellular therapies are being developed, including immunotherapy for cancer and infectious diseases, chondrocyte therapy for cartilage defects, neuronal cell therapy for neurodegenerative diseases, and stem cell therapy for numerous indications. Many of these therapies require the removal of unwanted, detrimental cells for full efficacy to be realized. Gene therapy is another active area of developing medicine that can influence the success of cellular therapy. Given the rapid advances in the understanding of the human genome, it is likely that many genes will be available for insertion into cells prior to transplantation into patients. However, obtaining efficient targeted delivery of genes into specific cells of interest has remained a difficult obstacle in the development of these therapies. In the treatment of cancer, it has been found that high-dose chemotherapy and/or radiation therapy can be used to selectively kill rapidly dividing cancer cells in the body. Unfortunately, several other cell types in the body are also rapidly dividing, and in fact, the dose-limiting toxicity for most anti-cancer therapies is the killing of HSCs and progenitor cells in the bone marrow. HSC transplantation was developed as a therapy to rescue the hematopoietic system following anti-cancer treatments. Upon infusion, the HSCs and progenitor cells within the transplant selectively home to the bone marrow and engraft. This process is monitored clinically through daily blood cell counts. Once blood counts return to acceptable levels, usually within 20 to 30 days, the patient is considered engrafted and is released from the hospital. HSC transplants have been traditionally performed with bone marrow, but mobilized peripheral blood (obtained via leukapheresis after growth factor or low-dose chemotherapy administration) has recently become the preferred source because it eliminates the need to harvest approximately one liter of bone marrow from the patient. In addition, HSCs from mobilized peripheral blood result in more rapid engraftment (8 to 15 days), leading to less critical patient care and earlier discharge from the hospital. HSC transplantation has become an established therapy for treating many diseases, such that over 45.000 procedures were performed worldwide in 1997. HSC transplantation may be performed using either donor cells (allogeneic), or patient cells that have been harvested and cryopreserved prior to administration of high- dose anti-cancer therapy (autologous). Autologous transplants are widely used for treating a variety of diseases including breast cancer, Hodgkin's and non-Hodgkin's lymphomas, neuroblastoma, and multiple myeloma. The number of autologous transplants currently outnumbers allogeneic transplants by approximately a 2: 1 ratio. This ratio is increasing further, mainly due to graft-versus-host disease (GVHD) complications associated with allogeneic transplants. One of the most significant problems with autologous transplants is the reintroduction of tumor cells to the patient along with the HSCs. because these tumor cells contribute to relapse of the original disease. As a tumor grows, tumor cells eventually leave the original tumor site and migrate through the bloodstream to other locations in the body. This process, called tumor metastasis, results in the formation and growth of satellite tumors that greatly increase the severity of the disease. The presence of these metastatic tumor cells in the blood and other tissues, often including bone marrow, can create a significant problem for autologous transplantation. In fact, there is a very high probability that metastatic tumor cells will contaminate the harvested HSCs that are to be returned to the patient following anti-cancer therapy. The presence of contaminating tumor cells in autologous bone marrow and mobilized peripheral blood harvests has been confirmed in numerous scientific studies. Recent landmark studies have unambiguously shown that reinfused tumor cells do indeed contribute to disease relapse in humans (Rill et al. 1994). This was proven by genetically marking the harvested cells prior to transplant, and then showing that the marker was detected in resurgent tumor cells in those patients who relapsed with disease. These data have been confirmed by other investigators (Deisseroth et al. 1994), indicating that contaminating tumor cells in HSC transplants represent a real threat to patients undergoing autologous transplantation. Subsequent detailed studies have now shown that the actual number of tumor cells reinfused in the transplant was correlated with the risk of relapse for acute lymphoblastic leukemia (Vervoordeldonk et al. 1997), non-Hodgkin's lymphoma (Sharp et al. 1992; Sharp et al. 1996), mantle cell lymphoma (Andersen et al. 1997), and breast cancer (Brockstein et al. 1996; Fields et al. 1996: Schulze et al. 1997; Vannucchi et al. 1998; Vredenburgh et al. 1997). One of these studies went even further, showing that the number of tumor cells infused was inversely correlated with the elapsed time to relapse (Vredenburgh et al. 1997). These data suggest that reducing the number of tumor cells in the transplant will lead to better outcomes for the patient. Due to the known risk of tumor cell contamination in autologous transplantation, a number of methods have been proposed for removing contaminating tumor cells from harvested HSC populations. The basic principle underlying all purging methods is to remove or kill tumor cells while preserving the HSCs that are needed for hematopoietic reconstitution in the patient. One such method utilized fluorescence-activated cell sorting (FACS) to sort HSCs from tumor cells (Tricot et al. 1995). As is known, flow cytometry sorts cells one at a time and physically separates one population of cells from a mixture of cells based upon cell surface markers and physical characteristics. However, it has been shown that using FACS to separate large cell populations for clinical applications is not advantageous because the process is slow, the cell yields can be very low, and purity greater than ~98% is rarely achieved. Another method utilizing a flow cytometer is described in U.S. Patent No. 4,395,397 to Shapiro. In the Shapiro method, labeled cells are placed in a flow cytometer, and a downstream laser beam is used to kill the labeled cells in the flowing stream after they pass by the detector and are recognized as being labeled by the electronic system. This method suffers from a number of disadvantages. Firstly, once an unwanted cell has passed through the detector/laser region there is no way to check that destruction has been completed successfully. If a tumor cell evades destruction it will inevitably be reintroduced into the patient. Secondly, the focal spot diameter of the laser beam is of necessity greater than the liquid stream cross section. Accordingly, many of the HSCs in the region of an unwanted cell will also be destroyed by the laser beam. Also, as described above, the purity obtained by flow cytometric techniques is not very good due to the random and dynamic nature of a heterogeneous cell mixture that is flowing in a fast-moving (1-20 m/sec) stream of liquid. Another method that utilizes laser technology is described in U.S. Patent No. 4,629,687 to Schindler, et al. In this method, anchorage-dependent cells are grown on a movable surface, and then a small laser beam spot is scanned across the moving surface to illuminate cells one at a time and the information is recorded. The same laser is then switched to a higher lethal power level, and the beam is swept over the surface in all areas except where a cell of interest was recorded during the illumination step. Unfortunately, this method is slow and only will work on cells that can adhere to a surface. A still further method that utilizes laser technology is described in U.S. Patent No. 5,035,693 to Kratzer. In this method, cells are placed on a moving belt and a small laser beam spot is scanned across the surface. When a particular cell radiates in response to the illuminating laser spot, the same laser is quickly switched to high power in order to kill the cell in a near simultaneous manner before the scanner has moved appreciably away from that cell. However, this system has many of the same disadvantages as the Shapiro method. For example, because the scanner is continuously moving during the imaging and killing of cells, the system is highly-dynamic, and therefore less stable and less accurate than a static system. Also, because the cells are moving on a belt past the detector in one direction, the method is not reversible. Thus, if a single tumor cell escapes detection, it will be reintroduced into the patient. Others have used a small laser beam spot to dynamically scan over a surface to illuminate cells. For example, U.S. Patent No. 4,284,897 to Sawamura et al. describes the use of galvanometric mirrors to scan a small laser beam spot in a standard microscope to illuminate fluorescent cells. U.S. Patent No. 5,381,224 to Dixon et al. describes imaging of macroscopic specimens through the use of a laser beam spot that is raster-scanned with galvanometric mirrors through an F-theta scanning lens. In U.S. Patent Nos. 5,646,411, 5,672,880, and 5,719,391 to Kain, scanning of a small laser spot with galvanometers through an F-theta lens is described. All of these imaging methods dynamically illuminate a small point that is moved over the surface to be imaged. In some cases, the surface being scanned is also moving during imaging. Similar methods of scanning a small laser spot have been described for purposes other than imaging of cells. For example, U.S. Patent No. 4,532,402 to Overbeck describes the use of galvanometers to move a small laser beam spot over a semiconductor surface for repair of an integrated circuit. Similarly, U.S. Patent No. 5,690,846 to Okada et al. describes laser processing by moving a small laser spot with mirrors through an F-theta scanning lens. U.S. Patent No. 5,296,963 to Murakami et al. describes the use of galvanometric mirrors to scan a small laser beam spot in a standard inverted microscope to puncture cells for insertion of genetic matter. Yet another method of scanning a biological specimen is described in U.S. Patent No. 5,932,872 to Price. This method uses a plurality of detectors to simultaneously capture images at a plurality of focus planes from a constantly moving surface. The resultant images can be used to choose the best-focus image in real-time, and can be used to generate a three-dimensional volumetric image of a specimen. Most of the methods described above are based on administering a tumor cell- removal or tumor cell-killing strategy to the entire harvested cell population as a whole. In flow cytometry, cells are sorted on a single cell basis to physically separate the unwanted tumor cells from HSCs. While each of these methods has been shown to reduce tumor cell numbers in HSC transplants, none has demonstrated the ability to remove or kill all detectable tumor cells. In fact, the majority of patient transplants still contain detectable tumor cells after these purging techniques are used. Approximately 30 to 30,000 tumor cells per transplant still remain, even after multiple-step purging procedures (Gazitt et al. 1995; Gribben et al. 1991; Mapara et al. 1997; Paulus et al. 1997). Further, all of these methods result in some degree of HSC loss or damage, which can significantly impact the success of the HSC transplant by delaying patient engraftment. In summary, existing purging technologies are inadequate, and there exists a great unmet clinical need for novel approaches that can effectively purge all detectable tumor cells from an HSC transplant. The method and apparatus described herein fulfills this need. High throughput screening for the action of candidate drug compounds on biological specimens is another area of great importance. Typically, a large number of candidate compounds is applied in parallel to small cell samples placed in wells of a multi- well plate, and each well is examined for some change in a biological indicator. Due to the large number of compounds, speed of screening is an important factor. Such studies are currently limited by capturing a signal from the cell population as a whole, or by laborious manual viewing of individual cells with microscopes. The former precludes the possibility of observing an effect on a cell subpopulation or of observing an effect within only a portion of the cell, whereas the latter approach is too slow to apply to numerous candidate compounds. An apparatus and method that could rapidly measure the effect of candidate compounds on individual cells is in great need. Summary of the Invention This invention provides a high-speed method and apparatus for selectively identifying, and individually targeting with an energy beam, specific cells within a cell population for the purpose of inducing a response in the targeted cells. Using the apparatus of the present invention, every detectable target cell in a cell population can be specifically identified and targeted, without substantially affecting cells that are not being targeted. The cells can be a mixed population or of a homogenous origin. Specific cells are identified with the disclosed invention using several approaches. One embodiment includes a non-destructive labeling method so that all of the cells of a first population are substantially distinguishable from the remaining cells of the cell mixture, the remaining cells comprising the second population. In this embodiment, a labeled antibody can be used to specifically mark each cell of the first population, yet not mark cells of the second population. The labeled cells are then identified within the cell mixture. A narrow energy beam is thereafter focused on the first of the targeted cells to achieve a desired response. The next of the targeted cells is then irradiated, and so on until every targeted cell has been irradiated. In another embodiment, an antibody that selectively binds to cells of the second population, but not cells of the first population, is used to identify cells of the first population. Cells of the first population are identified by the absence of the label, and are thereafter individually targeted with the energy beam. The nature of the response that is induced by the energy beam is dependent upon the nature of the energy beam. The response can be lethal or non-lethal. Thus, examples of responses that can be induced with an energy beam include necrosis, apoptosis, optoporation (to allow entry of a substance that is present in the surrounding medium, including genetic material), cell lysis, cell motion (laser tweezers), cutting of cell components (laser scissors), activation of a photosensitive substance, excitation of a fluorescent reagent, photobleaching, and molecular uncaging. Another aspect of the invention is directed to an apparatus for determining a morphological or physiological characteristic of individual cells in a biological specimen. Thus, the apparatus would directly or indirectly induce a non-lethal response in a population of cells, and measure the response. Such non-lethal responses include optoporation, cell motion (laser tweezers), cutting of cell components (laser scissors), activation of a photosensitive substance and excitation of a fluorescent reagent. Such non- lethal responses may also include the responses of cells targeted by the techniques of photobleaching, such as photobleaching recovery, and molecular uncaging, both internal and external with respect to the cell. An example of such apparatus would comprise: An illumination source for illuminating a frame of cells in said biological specimen; An image capture system that captures an image of said frame of cells; First commands for determining the location of a first individual cell in said biological specimen by reference to said image; An energy source that emits an energy beam sufficient to induce a response in at least one or more individual cells; Second commands for intersecting said first individual cell with an energy beam sufficient to interrogate said first individual cell for the presence of a morphological or physiological characteristic; and A detector for measuring the response of said first individual cell to said interrogation. Alternatively, such an apparatus would comprise: An illumination source for illuminating a frame of cells in said biological specimen; An image capture system that captures an image of said frame of cells; First commands for determining the location of a first targeted cell in said biological specimen by reference to said image; Second commands for intersecting said first targeted cell with an energy beam sufficient to cause a change of a morphological or physiological characteristic in said first targeted cell; and A detector for measuring the response for the change in said morphologic or physiological characteristic of said first targeted cell. Another aspect of the invention is an apparatus an illumination source for illuminating a frame of cells in said biological specimen comprising, an image capture system that captures an image of said frame of cells, first commands for determining the location of an individual cell in said biological specimen by reference to said image, an energy source that emits an energy beam sufficient to induce a response in at least one of the individual cells, second commands for intersecting said first individual cell with an energy beam sufficient to cause a change of a morphological or physiological characteristic in said first individual cell, and a detector for measuring a morphological or physiological characteristic of said first individual cell. Another aspect of the invention is an apparatus for determining a morphological or physiological characteristic of individual cells in a biological specimen comprising: an illumination source for illuminating a frame of cells in said biological specimen, wherein said specimen contains a quantity of caged compounds; an image capture system that captures an image of said frame of cells; first commands for intersecting one or more of said quantity of caged compounds with an energy beam sufficient to uncage said one or more of said quantity of caged compounds; and an energy source that emits an energy beam sufficient to uncage one or more of said quantities of caged compounds; second commands to monitor the progress of the change in said morphological or physiological characteristic in response to said uncaged compounds of said one or more of said quantity of caged compounds. Another such apparatus for determining a morphological or physiological characteristic of individual cells in a biological speciment comprises: an illumination source for illuminating a frame of cells in said biological specimen; an image capture system that captures an image of said frame of cells; and first comands for determining the morphological or physiological characteristic of said individual cell in said frame of cells. The above apparatus can be used for high throughput screening of responses of cells to outside stimuli, in that it provides rapid location and measurement of responses of individual cells to such stimuli. Thus, the illumination source and/or the energy beam could target molecules in the vicinity of the cells, whether or not the molecules are in or on the cells, the energy beam could interact with and activate the molecules and the illumination source could study the response of the cells to the molecules so activated. A large number of commercially important research and clinical applications can be envisioned for such an apparatus, examples of which are presented below. Brief Description of the Drawings FIGURE 1 is a perspective view of one embodiment of a cell treatment apparatus and illustrates the outer design of the housing and display. FIGURE 2 is a perspective view of one embodiment of a cell treatment apparatus with the outer housing removed and the inner components illustrated. FIGURE 3 is a block diagram of the optical subassembly design within one embodiment of a cell treatment apparatus. FIGURE 4 is a perspective view of one embodiment of an optical subassembly within one embodiment of a cell treatment apparatus. FIGURE 5 is a side view of one embodiment of an optical subassembly that illustrates the arrangement of the scanning lens and the movable stage. FIGURE 6 is a bottom perspective view of one embodiment of an optical subassembly. FIGURE 7 is a top perspective view of the movable stage of the cell treatment apparatus. Detailed Description A method and apparatus is described for selectively identifying, and individually targeting with an energy beam, specific cells within a cell population for the purpose of inducing a response in the targeted cells. The population of cells can be a mixed population or homogenous in origin. The responses of any of the embodiments of the methods and apparatuses of the invention can be lethal or non-lethal. Examples of such responses are set forth above and throughout this disclosure. The cells targeted can be labeled as is often the case when the specimen is a mixed population. On the other hand, when the specimen is homogenous, the targeted cells can be those individual cells that are being interrogated or intersected by the illumination source or the energy beam, in order to study the response of the cell. For instance, such responses include the morphological or physiological characteristics of the cell. Generally, the method first employs a label that acts as a marker to identify and locate individual cells of a first population of cells within a cell mixture that is comprised of the first population of cells and a second population of cells. The cells targeted by the apparatus and methods herein are those that are selectively labeled, in the case of a mixed population of cells, or the ones undergoing interrogation or intersection by the illumination source or energy beam. The chosen label can be any that substantially identifies and distinguishes the first population of cells from the second population of cells. For example, monoclonal antibodies that are directly or indirectly tagged with a fluorochrome can be used as specific labels. Other examples of cell surface binding labels include non-antibody proteins, lectins, carbohydrates, or short peptides with selective cell binding capacity. Membrane intercalating dyes, such as PKH-2 and PKH-26, could also serve as a useful distinguishing label indicating mitotic history of a cell. Many membrane-permeable reagents are also available to distinguish living cells from one another based upon selected criteria. For example, phalloidin indicates membrane integrity, tetramethyl rhodamine methyl ester (TMRM) indicates mitochondrial transmembrane potential, monochlorobimane indicates glutathione reductive stage, carboxymethyl fluoresce.n diacetate (CMFDA) indicates thiol activity, carboxyfluorescein diacetate indicates intracellular pH, fura-2 indicates intracellular Ca 2+ level, and 5,5',6,6'-tetrachloro-l,r,3,3'-tetraethylbenzimidazolo carbocyanine iodide (JC-1) indicates membrane potential. Cell viability can be assessed by the use of fluorescent SYTO 13 or YO PRO reagents. Similarly, a fluorescently-tagged genetic probe (DNA or RNA) could be used to label cells which carry a gene of interest, or express a gene of interest. Further, cell cycle status could be assessed through the use of Hoechst 33342 dye to label existing DNA combined with bromodeoxyuridine (BrdU) to label newly synthesized DNA. It should be noted that if no specific label is available for cells of the first population, the method can be implemented in an inverse fashion by utilizing a specific label for cells of the second population. For example, in hematopoietic cell populations, the CD34 or ACC-133 cell markers can be used to label only the primitive hematopoietic cells, but not the other cells within the mixture. In this embodiment, cells of the first population are identified by the absence of the label, and are thereby targeted by the energy beam. After cells of the first population are identified, an energy beam, such as from a laser, collimated or focused non-laser light, RF energy, accelerated particle, focused ultrasonic energy, electron beam, or other radiation beam, is used to deliver a targeted dose of energy that induces the pre-determined response in each of the cells of the first population, without substantially affecting cells of the second population. One such pre-determined response is photobleaching. In photobleaching, a label in the form of a dye, such as rhodamine 123, GFP, fluorescein isothiocyanate (FITC), or phycoerythrin, is added to the specimen before the instant methods are commenced. After the population of cells has time to interact with the dye, the energy beam is used to bleach a region of individual cells in the population. Such photobleaching studies can be used to study the motility, replenishment, dynamics and the like of cellular components and processes. Another response is internal molecular uncaging. In such a process, the specimen is combined with a caged molecule prior to the commencement of the instant methods. Such caged molecules include the beta-2,6-dinitrobenzyl ester of L-aspartic acid or the l-(2- nitrophenyl)ethyl ether of 8-hydroxypyrene-l,3,6-tris-sulfonic acid. Similarly, caging groups including alpha-carboxy-2-nitrobenzyl (CNB) and 5-carboxymethoxy-2-nitrobenzyl (CMNB) can be linked to biologically active molecules as ethers, thioethers, esters, amines, or similar functional groups. The term "internal molecular uncaging" refers to the fact that the molecular uncaging takes place on the surface or within the cell. Such uncaging experiments study rapid molecular processes such as cell membrane permeability and cellular signaling. Yet another response is external molecular uncaging. This uses approximately the same process as internal molecular caging. However, in external molecular uncaging, the uncaged molecule is not attached to or incorporated into the targeted cells. Instead, the responses of the surrounding targeted cells to the caged and uncaged variants of the molecule are imaged by the instant apparatus and methods. Figure 1 is an illustration of one embodiment of a cell treatment apparatus 10. The cell treatment apparatus 10 includes a housing 15 that stores the inner components of the apparatus. The housing includes laser safety interlocks to ensure safety of the user, and also limits interference by external influences (e.g., ambient light, dust, etc.). Located on the upper portion of the housing 15 is a display unit 20 for displaying captured images of cell populations during treatment. These images are captured by a camera, as will be discussed more specifically below. A keyboard 25 and mouse 30 are used to input data and control the apparatus 10. An access door 35 provides access to a movable stage that holds a specimen container of cells undergoing treatment. An interior view of the apparatus 10 is provided in Figure 2. As illustrated, the apparatus 10 provides an upper tray 200 and lower tray 210 that hold the interior components of the apparatus. The upper tray 200 includes a pair of intake filters 215A,B that filter ambient air being drawn into the interior of the apparatus 10. Below the access door 35 is the optical subassembly (not shown). The optical subassembly is mounted to the upper tray 200 and is discussed in detail with regard to Figures 3-6. On the lower tray 210 is a computer 225 which stores the software programs, commands and instructions that run the apparatus 10. In addition, the computer 225 provides control signals to the treatment apparatus through electrical signal connections for steering the laser to the appropriate spot on the specimen in order to treat the cells. As illustrated, a series of power supplies 230A,B,C provide power to the various electrical components within the apparatus 10. In addition, an uninterruptable power supply 235 is incorporated to allow the apparatus to continue functioning through short external power interruptions. Figure 3 provides a layout of one embodiment of an optical subassembly design 300 within an embodiment of a cell treatment apparatus 10. As illustrated, an illumination laser 305 provides a directed laser output that is used to excite a particular label that is attached to targeted cells within the specimen. In this embodiment, the illumination laser emits light at a wavelength of 532 nm. Once the illumination laser has generated a light beam, the light passes into a shutter 310 which controls the pulse length of the laser light. After the illumination laser light passes through the shutter 310, it enters a ball lens 315 where it is focused into a SMA fiber optic connector 320. After the illumination laser beam has entered the fiber optic connector 320, it is transmitted through a fiber optic cable 325 to an outlet 330. By passing the illumination beam through the fiber optic cable 325, the illumination laser 305 can be positioned anywhere within the treatment apparatus and thus is not limited to only being positioned within a direct light pathway to the optical components. In one embodiment, the fiber optic cable 325 is connected to a vibrating motor 327 for the purpose of mode scrambling and generating a more uniform illumination spot. After the light passes through the outlet 330, it is directed into a series of condensing lenses in order to focus the beam to the proper diameter for illuminating one frame of cells. As used herein, one frame of cells is defined as the portion of the biological specimen that is captured within one frame image captured by the camera. This is described more specifically below. Accordingly, the illumination laser beam passes through a first condenser lens 335. In one embodiment, this first lens has a focal length of 4.6 mm. The light beam then passes through a second condenser lens 340 which, in one embodiment, provides a 100 mm focal length. Finally, the light beam passes into a third condenser lens 345, which preferably provides a 200 mm focal length. While the present invention has been described using specific condenser lenses, it should be apparent that other similar lens configurations that focus the illumination laser beam to an advantageous diameter would function similarly. Thus, this invention is not limited to the specific implementation of any particular condenser lens system. Once the illumination laser beam passes through the third condenser lens 345, it enters a cube beamsplitter 350 that is designed to transmit the 532 nm wavelength of light emanating from the illumination laser. Preferably, the cube beamsplitter 350 is a 25.4 mm square cube (Melles-Griot, Irvine, CA). However, other sizes are anticipated to function similarly. In addition, a number of plate beamsplitters or pellicle beamsplitters could be used in place of the cube beamsplitter 350 with no appreciable change in function. Once the illumination laser light has been transmitted through the cube beamsplitter 350, it reaches a long wave pass mirror 355 that reflects the 532 nm illumination laser light to a set of galvanometer mirrors 360 that steer the illumination laser light under computer control to a scanning lens (Special Optics, Wharton, NJ) 365, which directs the illumination laser light to the specimen (not shown). The galvanometer mirrors are controlled so that the illumination laser light is directed at the proper cell population (i.e. frame of cells) for imaging. The "scanning lens" described in this embodiment of the invention includes a refractive lens. It should be noted that the term "scanning lens" as used in the present invention includes, but is not limited to, a system of one or more refractive or reflective optical elements used alone or in combination. Further, the "scanning lens" may include a system of one or more diffractive elements used in combination with one or more refractive and/or reflective optical elements. One skilled in the art will know how to design a "scanning lens" system in order to illuminate the proper cell population. The light from the illumination laser is of a wavelength that is useful for illuminating the specimen. In this embodiment, energy from a continuous wave 532 nm Nd:YAG frequency-doubled laser (B&W Tek, Newark, DE) reflects off the long wave pass mirror (Custom Scientific, Phoenix, AZ) and excites fluorescent tags in the specimen. In one embodiment, the fluorescent tag is phycoerythrin. Alternatively, Alexa 532 (Molecular Probes, Eugene, OR) can be used. Phycoerythrin and Alexa 532 have emission spectra with peaks near 580 nm, so that the emitted fluorescent light from the specimen is transmitted via the long wave pass mirror to be directed into the camera. The use of the filter in front of the camera blocks light that is not within the wavelength range of interest, thereby reducing the amount of background light entering the camera. It is generally known that many other devices could be used in this manner to illuminate the specimen, including, but not limited to, an arc lamp (e.g., mercury, xenon, etc.) with or without filters, a light-emitting diode (LED), other types of lasers, etc. Advantages of this particular laser include high intensity, relatively efficient use of energy, compact size, and minimal heat generation. It is also generally known that other fluorochromes with different excitation and emission spectra could be used in such an apparatus with the appropriate selection of illumination source, filters, and long and/or short wave pass mirrors. For example, allophycocyanin (APC) could be excited with a 633 nm HeNe illumination laser, and fluoroisothiocyanate (FITC) could be excited with a 488 nm Argon illumination laser. One skilled in the art could propose many other optical layouts with various components in order to achieve the objective of this invention. In addition to the illumination laser 305. an optional treatment laser 400 is present to irradiate the targeted cells once they have been identified by image analysis. Of course, in one embodiment, the treatment induces necrosis of targeted cells within the cell population. As shown, the treatment laser 400 outputs an energy beam of 523 nm that passes through a shutter 410. Although the exemplary laser outputs an energy beam having a 523 nm wavelength, other sources that generate energy at other wavelengths are also within the scope of the present invention. Once the treatment laser energy beam passes through the shutter 410, it enters a beam expander (Special Optics, Wharton, NJ) 415 which adjusts the diameter of the energy beam to an appropriate size at the plane of the specimen. Following the beam expander 415 is a half- wave plate 420 which controls the polarization of the beam. The treatment laser energy beam is then reflected off a mirror 425 and enters the cube beamsplitter 350. The treatment laser energy beam is reflected by 90° in the cube beamsplitter 350, such that it is aligned with the exit pathway of the illumination laser light beam. Thus, the treatment laser energy beam and the illumination laser light beam both exit the cube beamsplitter 350 along the same light path. From the cube beamsplitter 350, the treatment laser beam reflects off the long wave pass mirror 355, is steered by the galvanometers 360, thereafter contacts the scanning lens 365, and finally is focused upon a targeted cell within the specimen. Again, the "scanning lens" described in this embodiment includes a refractive lens. As previously mentioned, the term "scanning lens" includes, but is not limited to, a system of one or more refractive or reflective optical elements used alone or in combination. Further, the "scanning lens" may include one or more diffractive elements used in combination with one or more refractive and/or reflective elements. One skilled in the art will know how to design a "scanning lens" system in order to focus upon the targeted cell within the specimen It should be noted that a small fraction of the illumination laser light beam passes through the long wave pass mirror 355 and enters a power meter sensor (Gentec, Palo Alto, CA) 445. The fraction of the beam entering the power sensor 445 is used to calculate the level of power emanating from the illumination laser 305. In an analogous fashion, a small fraction of the treatment laser energy beam passes through the cube beamsplitter 350 and enters a second power meter sensor (Gentec, Palo Alto, CA) 446. The fraction of the beam entering the power sensor 446 is used to calculate the level of power emanating from the treatment laser 400. The power meter sensors are electrically linked to the computer system so that instructions/commands within the computer system capture the power measurement and determine the amount of energy that was emitted. The energy beam from the treatment laser is of a wavelength that is useful for achieving a response in the cells. In the example shown, a pulsed 523 nm Nd:YLF frequency-doubled laser is used to heat a localized volume containing the targeted cell, such that it is induced to die within a pre-determined period of time. The mechanism of death is dependent upon the actual temperature achieved in the cell, as reviewed by Niemz (Niemz 1996). A Nd:YLF frequency-doubled, solid-state laser (Spectra-Physics. Mountain View, CA) is used because of its stability, high repetition rate of firing, and long time of maintenance-free service. However, most cell culture fluids and cells are relatively transparent to light in this green wavelength, and therefore a very high fluence of energy would be required to achieve cell death. To significantly reduce the amount of energy required, and therefore the cost and size of the treatment laser, a dye is purposefully added to the specimen to efficiently absorb the energy of the treatment laser in the specimen. In the example shown, the non-toxic dye FD&C red #40 (allura red) is used to absorb the 523 nm energy from the treatment laser, but one skilled in the art could identify other laser/dye combinations that would result in efficient absorption of energy by the specimen. For example, a 633 nm HeNe laser ' s energy would be efficiently absorbed by FD&C green #3 (fast green FCF), a 488 nm Argon laser's energy would be efficiently absorbed by FD&C yellow #5 (sunset yellow FCF), and a 1064 nm Nd:YAG laser's energy would be efficiently absorbed by Filtron (Gentex, Zeeland, MI) infrared absorbing dye. Through the use of an energy absorbing dye, the amount of energy required to kill a targeted cell can be reduced since more of the treatment laser energy is absorbed in the presence of such a dye. Another method of achieving thermal killing of cells without the addition of a dye involves the use of an ultraviolet laser. Energy from a 355 nm Nd:YAG frequency-tripled laser will be absorbed by nucleic acids and proteins within the cell, resulting in thermal heating and death. Yet another method of achieving thermal killing of cells without the addition of a dye involves the use of a near-infrared laser. Energy from a 2100 nm Ho:YAG laser or a 2940 nm Er:YAG laser will be absorbed by water within the cell, resulting in thermal heating and death. Although this embodiment describes the killing of cells via thermal heating by the energy beam, one skilled in the art would recognize that other responses can also be induced in the cells by an energy beam, including photomechanical disruption, photodissociation, photoablation, and photochemical reactions, as reviewed by Niemz (Niemz 1996). For example, a photosensitive substance (e.g., hemtoporphyrin derivative, tin-etiopurpurin, lutetuim texaphyrin) (Oleinick and Evans 1998) within the cell mixture could be specifically activated in targeted cells by irradiation. Additionally, a small, transient pore could be made in the cell membrane (Palumbo et al. 1996) to allow the entry of genetic or other material. Further, specific molecules in or on the cell, such as proteins or genetic material, could be inactivated by the directed energy beam (Grate and Wilson 1999; Jay 1988). Also, photobleaching can be utilized to measure intracellular movements such as the diffusion of proteins in membranes and the movements of microtubules during mitosis (Ladha et al., 1997 J. Cell Sci 1997 110(9): 1041; Centonze and Borisy, 1991 J. Cell Sci 100 (pt 1):205; White and Stelzer, Trends Cell Biol 1999 Feb; 9(2):61-5; Meyvis, et al., Pharm Res 1999 Aug; 16(8): 1 153-62). Further, photolysis or uncaging, including multiphoton uncaging, of caged compounds can be utilized to control the release, with temporal and spacial resolution, of biologically active products or other products of interest (Theriot and Mitchison, 1992 J. Cell Biol. 1 19:367; Denk, 1994 PNAS 91(14):6629). These mechanisms of inducing a response in a targeted cell via the use of electromagnetic radiation directed at specific targeted cells are also intended to be incorporated into the present invention. In addition to the illumination laser 305 and treatment laser 400, the apparatus includes a camera 450 that captures images (i.e. frames) of the cell populations. As illustrated in Figure 3, the camera 450 is focused through a lens 455 and filter 460 in order to accurately record an image of the cells without capturing stray background images. A stop 462 is positioned between the filter 460 and mirror 355 in order to eliminate light that may enter the camera from angles not associated with the image from the specimen. The filter 460 is chosen to only allow passage of light within a certain wavelength range. This wavelength range includes light that is emitted from the targeted cells upon excitation by the illumination laser 305, as well as light from a back-light source 475. The back-light source 475 is located above the specimen to provide back- illumination of the specimen at a wavelength different from that provided by the illumination laser 305. This LED generates light at 590 nm, such that it can be transmitted through the long wave pass mirror to be directed into the camera. This back-illumination is useful for imaging cells when there are no fluorescent targets within the frame being imaged. An example of the utility of this back-light is its use in attaining proper focus of the system, even when there are only unstained, non-fluorescent cells in the frame. In one embodiment, the back-light is mounted on the underside of the access door 35 (Figure 2). Thus, as discussed above, the only light returned to the camera is from wavelengths that are of interest in the specimen. Other wavelengths of light do not pass through the filter 460, and thus do not become recorded by the camera 450. This provides a more reliable mechanism for capturing images of only those cells of interest. It is readily apparent to one skilled in the art that the single filter 460 could be replaced by a movable filter wheel that would allow different filters to be moved in and out of the optical pathway. In such an embodiment, images of different wavelengths of light could be captured at different times during cell processing, allowing the use of multiple cell labels. It should be noted that in this embodiment, the camera is a charge-coupled device (CCD) and transmits images back to the computer system for processing. As will be described below, the computer system determines the coordinates of the targeted cells in the specimen by reference to the image captured by the CCD camera. Referring now to Figure 4, a perspective view of an embodiment of an optical subassembly is illustrated. As illustrated, the illumination laser 305 sends a light beam through the shutter 310 and ball lens 315 to the SMA fiber optic connector 320. The light passes through the fiber optic cable 325 and through the output 330 into the condenser lenses 335, 340 and 345. The light then enters the cube beamsplitter 350 and is transmitted to the long wave pass mirror 355. From the long wave pass mirror 355, the light beam enters the computer-controlled galvanometers 360 and is then steered to the proper frame of cells in the specimen from the scanning lens 365. As also illustrated in the perspective drawing of Figure 4, the treatment laser 400 transmits energy through the shutter 410 and into the beam expander 415. Energy from the treatment laser 400 passes through the beam expander 415 and passes through the half- wave plate 420 before hitting the fold mirror 425, entering the cube beamsplitter 350 where it is reflected 90° to the long wave pass mirror 355, from which it is reflected into the computer controlled galvanometer mirrors 360. After being steered by the galvanometer mirrors 360 to the scanning lens 365, the laser energy beam strikes the proper location within the cell population in order to induce a response in a particular targeted cell. In order to accommodate a very large surface area of specimen to treat, the apparatus includes a movable stage that mechanically moves the specimen container with respect to the scanning lens. Thus, once a specific sub-population (i.e. field) of cells within the scanning lens field-of-view has been treated, the movable stage brings another sub- population of cells within the scanning lens field-of-view. As illustrated in Figure 5, a computer-controlled movable stage 500 holds a specimen container (not shown) to be processed. The movable stage 500 is moved by computer-controlled servo motors along two axes so that the specimen container can be moved relative to the optical components of the instrument. The stage movement along a defined path is coordinated with other operations of the apparatus. In addition, specific coordinates can be saved and recalled to allow return of the movable stage to positions of interest. Encoders on the x and y movement provide closed-loop feedback control on stage position. The flat-field (F-theta) scanning lens 365 is mounted below the movable stage. The scanning lens field-of-view comprises the portion of the specimen that is presently positioned above the scanning lens by the movable stage 500. The lens 365 is mounted to a stepper motor that allows the lens 365 to be automatically raised and lowered (along the z- axis) for the purpose of focusing the system. As illustrated in Figures 4-6, below the scanning lens 365 are the galvanometer- controlled steering mirrors 360 that deflect electromagnetic energy along two perpendicular axes. Behind the steering mirrors is the long wave pass mirror 355 that reflects electromagnetic energy of a wavelength shorter than 545 nm. Wavelengths longer than 545 nm are passed through the long wave pass mirror, directed through the filter 460, coupling lens 455, and into the CCD camera, thereby producing an image of the appropriate size on the CCD sensor of the camera 450 (See Figures 3 and 4). The magnification defined by the combination of the scanning lens 365 and coupling lens 455 is chosen to reliably detect single cells while maximizing the area viewed in one frame by the camera. Although a CCD camera (DVC, Austin, TX) is illustrated in this embodiment, the camera can be any type of detector or image gathering equipment known to those skilled in the art. The optical subassembly of the apparatus is preferably mounted on a vibration-isolated platform to provide stability during operation as illustrated in Figures 2 and 5. Referring now to Figure 7, a top view of the movable stage 500 is illustrated. As shown, a specimen container is mounted in the movable stage 500. The specimen container 505 rests on an upper axis nest plate 510 that is designed to move in the forward/backward direction with respect to the movable stage 500. A stepper motor (not shown) is connected to the upper axis nest plate 510 and computer system so that commands from the computer cause forward/backward movement of the specimen container 505. The movable stage 500 is also connected to a timing belt 515 that provides side-to- side movement of the movable stage 500 along a pair of bearing tracks 525A,B. The timing belt 515 attaches to a pulley (not shown) housed under a pulley cover 530. The pulley is connected to a stepper motor 535 that drives the timing belt 515 to result in side- to-side movement of the movable stage 500. The stepper motor 535 is electrically connected to the computer system so that commands within the computer system result in side-to-side movement of the movable stage 500. A travel limit sensor 540 connects to the computer system and causes an alert if the movable stage travels beyond a predetermined lateral distance. A pair of accelerometers 545A,B is preferably incorporated on this platform to register any excessive bumps or vibrations that may interfere with the apparatus operation. In addition, a two-axis inclinometer 550 is preferably incorporated on the movable stage to ensure that the specimen container is level, thereby reducing the possibility of gravity- induced motion in the specimen container. The specimen chamber has a fan with ductwork to eliminate condensation on the specimen container, and a thermocouple to determine whether the specimen chamber is within an acceptable temperature range. Additional fans are provided to expel the heat generated by the electronic components, and appropriate filters are used on the air intakes 215A.B. The computer system 225 controls the operation and synchronization of the various pieces of electronic hardware described above. The computer system can be any commercially available computer that can interface with the hardware. One example of such a computer system is an Intel Pentium II. Ill or IV-based computer running the Microsoft Windows ® NT operating system. Software is used to communicate with the various devices, and control the operation in the manner that is described below. When the apparatus is first initialized, the computer loads files from the hard drive into RAM for proper initialization of the apparatus. A number of built-in tests are automatically performed to ensure the apparatus is operating properly, and calibration routines are executed to calibrate the apparatus. Upon successful completion of these routines, the user is prompted to enter information via the keyboard and mouse regarding the procedure that is to be performed. Once the required information is entered, the user is prompted to open the access door 35 and load a specimen onto the movable stage. Once a specimen is in place on the movable stage and the door is closed, the computer passes a signal to the stage to move into a home position. The fan is initialized to begin warming and defogging of the specimen. During this time, cells within the specimen are allowed to settle to the bottom surface. In addition, during this time, the apparatus may run commands that ensure that the specimen is properly loaded, and is within the focal range of the system optics. For example, specific markings on the specimen container can be located and focused on by the system to ensure that the scanning lens has been properly focused on the bottom of the specimen container. Such markings could also be used by the instrument to identify the container, its contents, and even the procedure to be performed. After a suitable time, the computer turns off the fan to prevent excess vibrations during treatment, and cell processing begins. First, the computer instructs the movable stage to be positioned over the scanning lens so that the first area (i.e. field) of the specimen to be treated is directly in the scanning lens field-of-view. The galvanometer mirrors are instructed to move such that the center frame within the field-of-view is imaged in the camera. As discussed below, the field imaged by the scanning lens is separated into a plurality of frames. Each frame is the proper size so that the cells within the frame are effectively imaged by the camera. The back-light 475 is then activated in order to illuminate the field-of-view so that it can be brought into focus by the scanning lens. Once the scanning lens has been properly focused upon the specimen, the computer system divides the field-of-view into a plurality of frames so that each frame is analyzed separately by the camera. This methodology allows the apparatus to process a plurality of frames within a large field-of-view without moving the mechanical stage. Because the galvanometers can move from one frame to the next very rapidly compared to the mechanical steps involved in moving the stage, this method results is an extremely fast and efficient apparatus. Other means of ensuring that the specimen is in focus are also available. For example, a laser proximeter (Cooke Corp., Auburn. MI) could rapidly determine the distance between the scanning lens and the sample, and adjust the scanning lens position accordingly. Ultrasonic proximeters are also available, and would achieve the same objective. One skilled in the art could propose other means of ensuring that the specimen is in focus above the scanning lens. In one preferred embodiment, the apparatus described herein processes at least 1 , 2, 3, 4, 5, 6, 7, or 14 square centimeters of a biological specimen per minute. In another embodiment, the apparatus described herein processes at least 0.25, 0.5, 1, 2, 3, 4 or 8 million cells of a biological specimen per minute. In one other embodiment, the apparatus can preferably induce a response in targeted cells at a rate of 50, 100, 150, 200, 250, 300, 350, 400 or 800 cells per second. Initially, an image of the frame at the center of the field-of-view is captured by the camera and stored to a memory in the computer. Instructions in the computer analyze the focus of the specimen by looking at the size of, number of, and other object features in the image. If necessary, the computer instructs the z-axis motor attached to the scanning lens to raise or lower in order to achieve the best focus. The apparatus may iteratively analyze the image at several z-positions until the best focus is achieved. The galvanometer- controlled mirrors are then instructed to image a first frame, within the field-of-view, in the camera. For example, the entire field-of-view might be divided into 4, 9, 12, 18, 24 or more separate frames that will be individually captured by the camera. Once the galvanometer mirrors are pointed to the first frame in the field-of-view, the shutter in front of the illumination laser is opened to illuminate the first frame through the galvanometer mirrors and scanning lens. The camera captures an image of any fluorescent emission from the specimen in the first frame of cells. Once the image has been acquired, the shutter in front of the illumination laser is closed and a software program (Epic. Buffalo Grove, IL) within the computer processes the image. The power sensor 445 discussed above detects the level of light that was emitted by the illumination laser, thereby allowing the computer to calculate if it was adequate to illuminate the frame of cells. If not, another illumination and image capture sequence is performed. Repeated failure to sufficiently illuminate the specimen will result in an error condition that is communicated to the operator. Shuttering of illumination light reduces undesirable heating and photobleaching of the specimen and provides a more repeatable fluorescent signal. An image analysis algorithm is run to locate the x-y centroid coordinates of all targeted cells in the frame by reference to features in the captured image. If there are targets in the image, the computer calculates the two-dimensional coordinates of all target locations in relation to the movable stage position and field-of-view, and then positions the galvanometer-controlled mirrors to point to the location of the first target in the first frame of cells. It should be noted that only a single frame of cells within the field-of-view has been captured and analyzed at this point. Thus, there should be a relatively small number of identified targets within this sub- population of the specimen. Moreover, because the camera is pointed to a smaller population of cells, a higher magnification is used so that each target is imaged by many pixels within the CCD camera. Once the computer system has positioned the galvanometer controlled mirrors to point to the location of the first targeted cell within the first frame of cells, the treatment laser is fired for a brief interval so that the first targeted cell is given an appropriate dose of energy. The power sensor 446 discussed above detects the level of energy that was emitted by the treatment laser, thereby allowing the computer to calculate if it was adequate to induce a response in the targeted cell. If not sufficient, the treatment laser is fired at the same target again. If repeated shots do not deliver the required energy dose, an error condition is communicated to the operator. These targeting, firing, and sensing steps are repeated by the computer for all targets identified in the captured frame. Once all of the targets have been irradiated with the treatment laser in the first frame of cells, the mirrors are then positioned to the second frame of cells in the field-of-view, and the processing repeats at the point of frame illumination and camera imaging. This processing continues for all frames within the field-of-view above the scanning lens. When all of these frames have been processed, the computer instructs the movable stage to move to the next field-of-view in the specimen, and the process repeats at the back-light illumination and auto-focus step. Frames and fields-of-view are appropriately overlapped to reduce the possibility of inadvertently missing areas of the specimen. Once the specimen has been fully processed, the operator is signaled to remove the specimen, and the apparatus is immediately ready for the next specimen. Although the text above describes the analysis of fluorescent images for locating targets, one can easily imagine that the non-fluorescent back-light LED illumination images will be useful for locating other types of targets as well, even if they are unlabeled. The advantage of using the galvanometer mirrors to control the imaging of successive frames and the irradiation of successive targets is significant. One brand of galvanometer is the Cambridge Technology, Inc. model number 6860 (Cambridge, MA). This galvanometer can reposition very accurately within a few milliseconds, making the processing of large areas and many targets possible within a reasonable amount of time. In contrast, the movable stage is relatively slow, and is therefore used only to move specified areas of the specimen into the scanning lens field-of-view. Error signals continuously generated by the galvanometer control boards are monitored by the computer to ensure that the mirrors are in position and stable before an image is captured, or before a target is fired upon, in a closed-loop fashion. In the context of the present invention, the term "specimen" has a broad meaning. It is intended to encompass any type of biological sample placed within the apparatus. The specimen may be enclosed by, or associated with, a container to maintain the sterility and viability of the cells. Further, the specimen may incorporate, or be associated with, a cooling apparatus to keep it above or below ambient temperature during operation of the methods described herein. The specimen container, if one is used, must be compatible with the use of the illumination laser, back-light illuminator, and treatment laser, such that it transmits adequate energy without being substantially damaged itself. Of course, many variations of the above-described embodiment are possible, including alternative methods for illuminating, imaging, and targeting the cells. For example, movement of the specimen relative to the scanning lens could be achieved by keeping the specimen substantially stationary while the scanning lens is moved. Steering of the illumination beam, images, and energy beam could be achieved through any controllable reflective or diffractive device, including prisms, piezo-electric tilt platforms, or acousto-optic deflectors. Additionally, the apparatus can image/process from either below or above the specimen. Because the apparatus is focused through a movable scanning lens, the illumination and energy beams can be directed to different focal planes along the z-axis. Thus, portions of the specimen that are located at different vertical heights can be specifically imaged and processed by the apparatus in a three-dimensional manner. The sequence of the steps could also be altered without changing the process. For example, one might locate and store the coordinates of all targets in the specimen, and then return to the targets to irradiate them with energy one or more times over a period of time. To optimally process the specimen, it should be placed on a substantially flat surface so that a large portion of the specimen appears within a narrow range of focus, thereby reducing the need for repeated auto-focus steps. The density of cells on this surface can, in principle, be at any value. However, the cell density should be as high as possible to minimize the total surface area required for the procedure. The following examples illustrate the use of the described method and apparatus in different applications. Example 1 : Autologous HSC Transplantation A patient with a B cell-derived metastatic tumor in need of an autologous HSC transplant is identified by a physician. As a first step in the treatment, the patient undergoes a standard HSC harvest procedure, resulting in collection of approximately 1 x 10 10 hematopoietic cells with an unknown number of contaminating tumor cells. The harvested cells are enriched for HSC by a commercial immunoaffinity column (Isolex ® 300, Nexell Therapeutics, Irvine, CA) that selects for cells bearing the CD34 surface antigen, resulting in a population of approximately 3 x 10 s hematopoietic cells, with an unknown number of tumor cells. The mixed population is thereafter contacted with anti-B cell antibodies (directed against CD20 and CD22) that are conjugated to phycoerythrin. The labeled antibodies specifically bind to the B cell-derived tumor cells. The mixed cell population is then placed in a sterile specimen container on a substantially flat surface near confluence, at approximately 500,000 cells per square centimeter. The specimen is placed on the movable stage of the apparatus described above, and all detectable tumor cells are identified by reference to phycoerythrin and targeted with a lethal dose of energy from a treatment laser. The design of the apparatus allows the processing of a clinical-scale transplant specimen in under 4 hours. The cells are recovered from the specimen container, washed, and then cryopreserved. Before the cells are reinfused, the patient is given high-dose chemotherapy to destroy the tumor cells in the patient's body. Following this treatment, the processed cells are thawed at 37 °C and are given to the patient intravenously. The patient subsequently recovers with no remission of the original cancer. Example 2: Allogeneic HSC Transplantation In another embodiment, the significant risk and severity of graft- versus-host disease in the allogeneic HSC transplant setting can be combated. A patient is selected for an allogeneic transplant once a suitable donor is found. Cells are harvested from the selected donor as described in the above example. In this case, the cell mixture is contacted with phycoerythrin-labeled anti-CD3 T-cell antibodies. Alternatively, specific allo-reactive T- cell subsets could be labeled using an activated T-cell marker (e.g. CD69) in the presence of allo-antigen. The cell population is processed by the apparatus described herein, thereby precisely defining and controlling the number of T-cells given to the patient. This type of control is advantageous, because administration of too many T-cells increases the risk of graft- versus-host disease, whereas too few T-cells increases the risk of graft failure and the risk of losing of the known beneficial graft- versus-leukemia effect. The present invention and methods are capable of precisely controlling the number of T-cells in an allogeneic transplant. Example 3: Tissue Engineering In another application, the present apparatus is used to remove contaminating cells in inocula for tissue engineering applications. Cell contamination problems exist in the establishment of primary cell cultures required for implementation of tissue engineering applications, as described by Langer and Vacanti (Langer and Vacanti 1999). In particular, chondrocyte therapies for cartilage defects are hampered by impurities in the cell populations derived from cartilage biopsies. Accordingly, the present invention is used to specifically remove these types of cells from the inocula. For example, a cartilage biopsy is taken from a patient in need of cartilage replacement. The specimen is then grown under conventional conditions (Brittberg et al. 1994). The culture is then stained with a specific label for any contaminating cells, such as fast-growing fibroblasts. The cell mixture is then placed within the apparatus described and the labeled, contaminating cells are targeted by the treatment laser, thereby allowing the slower growing chondrocytes to fully develop in culture. Example 4: Stem Cell Therapy Yet another embodiment involves the use of embryonic stem cells to treat a wide variety of diseases. Since embryonic stem cells are undifferentiated, they can be used to generate many types of tissue that would find use in transplantation, such as cardiomyocytes and neurons. However, undifferentiated embryonic stem cells that are implanted can also lead to a jumble of cell types which form a type of tumor known as a teratoma (Pedersen 1999). Therefore, therapeutic use of tissues derived from embryonic stem cells must include rigorous purification of cells to ensure that only sufficiently differentiated cells are implanted. The apparatus described herein is used to eliminate undifferentiated stem cells prior to implantation of embryonic stem cell-derived tissue in the patient. Example 5: Generation of Human Tumor Cell Cultures In another embodiment, a tumor biopsy is removed from a cancer patient for the purpose of initiating a culture of human tumor cells. However, the in vitro establishment of primary human tumor cell cultures from many tumor types is complicated by the presence of contaminating primary cell populations that have superior in vitro growth characteristics over tumor cells. For example, contaminating fibroblasts represent a major challenge in establishing many cancer cell cultures. The disclosed apparatus is used to particularly label and destroy the contaminating cells, while leaving the biopsied tumor cells intact. Accordingly, the more aggressive primary cells will not overtake and destroy the cancer cell line. Example 6: Generation of a Specific mRNA Expression Library The specific expression pattern of genes within different cell populations is of great interest to many researchers, and many studies have been performed to isolate and create libraries of expressed genes for different cell types. For example, knowing which genes are expressed in tumor cells versus normal cells is of great potential value (Cossman et al. 1999). Due to the amplification methods used to generate such libraries (e.g. PCR), even a small number of contaminating cells will result in an inaccurate expression library (Cossman et al. 1999; Schutze and Lahr 1998). One approach to overcome this problem is the use of laser capture microdissection (LCM), in which a single cell is used to provide the starting genetic material for amplification (Schutze, Lahr 1998). Unfortunately, gene expression in single cells is somewhat stochastic, and may be biased by the specific state of that individual cell at the time of analysis (Cossman et al. 1999). Therefore, accurate purification of a significant cell number prior to extraction of mRNA would enable the generation of a highly accurate expression library, one that is representative of the cell population being studied, without biases due to single cell expression or expression by contaminating cells. The methods and apparatus described in this invention can be used to purify cell populations so that no contaminating cells are present during an RNA extraction procedure. Example 7: Transfection of a Specific Cell Population Many research and clinical gene therapy applications are hampered by the inability to transfect an adequate number of a desired cell type without transfecting other cells that are present. The method of the present invention would allow selective targeting of cells to be transfected within a mixture of cells. By generating a photomechanical shock wave at or near a cell membrane with a targeted energy source, a transient pore can be formed, through which genetic (or other) material can enter the cell. This method of gene transfer has been called optoporation (Palumbo et al. 1996). The apparatus described above can achieve selective optoporation on only the cells of interest in a rapid, automated, targeted manner. For example, white blood cells are plated in a specimen container having a solution containing DNA to be transfected. Fluorescently-labeled antibodies having specificity for stem cells are added into the medium and bind to the stem cells. The specimen container is placed within the cell processing apparatus and a treatment laser is targeted to any cells that become fluorescent under the illumination laser light. The treatment laser facilitates transfection of DNA specifically into the targeted cells. Example 8: Selection of Desirable Clones in a Biotechnology Application In many biotechnology processes where cell lines are used to generate a valuable product, it is desirable to derive clones that are very efficient in producing the product. This selection of clones is often carried out manually, by inspecting a large number of clones that have been isolated in some manner. The present invention would allow rapid, automated inspection and selection of desirable clones for production of a particular product. For example, hybridoma cells that are producing the greatest amounts of antibody can be identified by a fluorescent label directed against the F c region. Cells with no or dim fluorescent labeling are targeted by the treatment laser for killing, leaving behind the best producing clones for use in antibody production. Example 9: Automated Monitoring of Cellular Responses Automated monitoring of cellular responses to specific stimuli is of great interest in high-throughput drug screening. Often, a cell population in one well of a well-plate is exposed to a stimulus, and a fluorescent signal is then captured over time from the cell population as a whole. Using the methods and apparatus described herein, more detailed monitoring could be done at the single cell level. For example, a cell population can be labeled to identify a characteristic of a subpopulation of cells that are of interest. This label is then excited by the illumination laser to identify those cells. Thereafter, the treatment laser is targeted at the individual cells identified by the first label, for the purpose of exciting a second label, thereby providing information about each cell's response. Since the cells are substantially stationary on a surface, each cell could be evaluated multiple times, thereby providing temporal information about the kinetics of each cell's response. Also, through the use of the large area scanning lens and galvanometer mirrors, a relatively large number of wells could be quickly monitored over a short period of time. As a specific example, consider the case of alloreactive T-cells as presented in Example 2 above. In the presence of allo-antigen, activated donor T-cells could be identified by CD69. Instead of using the treatment laser to target and kill these cells, the treatment laser could be used to examine the intracellular pH of every activated T-cell through the excitation and emitted fluorescence of carboxyfluorescein diacetate. The targeted laser allows the examination of only cells that are activated, whereas most screening methods evaluate the response of an entire cell population. If a series of such wells are being monitored in parallel, various agents could be added to individual wells, and the specific activated T-cell response to each agent could be monitored over time. Such an apparatus would provide a high-throughput screening method for agents that ameliorate the alloreactive T-cell response in graft-versus-host disease. Based on this example, one skilled in the art could imagine many other examples in which a cellular response to a stimulus is monitored on an individual cell basis, focusing only on cells of interest identified by the first label. Example 10: Photobleaching Studies Photobleaching, and/or photobleach recovery, of a specific area of a fluorescently- stained biological sample is a common method that is used to assess various biological processes. For example, a cell suspension is labeled with rhodamine 123, which fluorescently stains mitochondria within the cells. Using the instant illumination laser, the mitochondria within one or more cells are visualized due to rhodamine 123 fluorescence. The treatment laser is then used to deliver a focused beam of light that results in photobleaching of the rhodamine 123 in a small area within one or more cells. The photobleached area(s) then appear dark immediately thereafter, whereas adjacent areas are unaffected. A series of images are then taken using the illumination laser, providing a time-lapse series of images that document the migration of unbleached mitochondria into the area that was photobleached with the treatment laser. This approach can be used to assess the motion, turnover, or replenishment of many biological structures within cells. Thus, in cultured rat neurites, the photobleach recovery of mitochondria is a measure of the size of the mobile pool of mitochondria within each cell (Chute, et al. 1995). The rate of photobleach recovery in these cells is dependent on intracellular calcium and magnesium concentrations, energy status, and microtubule integrity. Neurotoxic substances, such as taxol or vinblastine, will affect the rate of photobleach recovery. Therefore, an assay for neurotoxic substances could be based on the measurement of photobleach recovery of mitochondria within a statistically significant number of neurites that had been exposed to various agents in the wells of a multi-well plate. In such an application, the apparatus described herein and used as described above, would provide a rapid automated method to assess neurotoxicity of many substances on a large number of cells. Based on this example, one skilled in the art could imagine many other examples in which photobleaching is induced and photobleach recovery is monitored in order to obtain useful information from a biological specimen. Example 1 1 : Uncaging Studies Use of caged compounds to study rapid biological processes involves the binding (i.e. caging) of a biologically relevant substance in an inactive state, allowing the caged substance to diffuse into the biological specimen (a relatively slow process), and then using a laser to induce a photolysis reaction (a relatively fast process) which liberates (i.e. uncages) the substance in situ over microsecond time scales. The biological specimen is then observed in short time-lapse microscopy in order to determine the effect of the uncaged substance on some biological process. Cages for many important substances have been described, including Dioxygen, cyclic ADP-ribose (cADPR), nicotinic acid adenine dinucleotide phosphate (NAADP), nitric oxide (NO), calcium, L-aspartate, and adenosine triphosphate (ATP). Chemotaxis is one example of a physiological characteristic that can be studied by uncaging compounds. Uncaging studies involve the irradiation of a portion of a biological specimen with laser light followed by examination of the specimen with time-lapse microscopy. The apparatus of the current invention has clear utility in such studies. As a specific example, consider the study of E. coli chemotaxis towards L-aspartate (Jasuja, et al. 1999). The beta- 2,6-dinitrobenzyl ester of L-aspartic acid and the 1 -(2-nitrophenyl)ethyl ether of 8- hydroxypyrene-l,3,6-tris-sulfonic acid are added to the wells of a well plate containing E. coli. Upon irradiation with the treatment laser, a localized uncaging of L-aspartate and the fluorophore 8-hydroxypyrene-l,3,6-tris-sulfonic acid (pyranine) is induced. The L- aspartate acts as a chemoattractant for E. coli., and in subsequent fluorescent images (using the illumination laser) the pyranine fluorophore acts as an indicator of the degree of uncaging that has occurred in the local area of irradiation. Time-lapse images of the E. coli. in the vicinity illuminated by visible wavelength light, such as from the back-light, of the uncaging event are used to measure the chemotactic response of the microorganisms to the locally uncaged L-aspartate. Due to the nature of the present invention, a large number of wells, each with a potential anti-microbial agent added, are screened in rapid order to determine the chemotactic response of microorganisms. Based on this example, one skilled in the art could imagine many other examples in which uncaging is induced by the treatment laser, followed by time-lapse microscopy in order to obtain useful information on a large number of samples in an automated fashion. Although aspects of the present invention have been described by particular embodiments exemplified herein, the present invention is not so limited. The present invention is only limited by the claims appended below. 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Claims: WE CLAIM: 1. An apparatus for selectively inducing a response in one or more targeted cells within a biological specimen, comprising: an illumination source for illuminating a frame of cells in said biological specimen; an image capture system that captures an image of said frame of cells; first commands for determining the locations of said one or more targeted cells within said image; an energy source that emits an energy beam sufficient to induce a response in at least one of said one or more targeted cells within said frame of cells; and second commands for steering said energy beam to said locations of said one or more targeted cells. 2. The apparatus of Claim 1. wherein said one or more targeted cells are identified by the presence of, or absence of, a label. 3. The apparatus of Claim 2. wherein said label is directed at the cell surface. 4. The apparatus of Claim 3, wherein said label comprises an antibody. 5. The apparatus of Claim 2, wherein said label comprises a membrane permeable reagent. 6. The apparatus of Claim 1, wherein said illumination source comprises a diffused laser light. 7. The apparatus of Claim 1 , wherein said illumination source comprises an arc lamp. 8. The apparatus of Claim 1, wherein said illumination source comprises a light-emitting diode. 9. The apparatus of Claim 1, wherein said illumination source comprises an incandescent lamp. 10. The apparatus of Claim 1, wherein said illumination source comprises a fluorescent lamp. 11. The apparatus of Claim 1 , wherein said illumination source only illuminates a sub-population of cells in said biological specimen. 12. The apparatus of Claim 1, wherein said one or more targeted cells comprises tumor cells. 13. The apparatus of Claim 1, wherein said one or more targeted cells comprises fibroblasts. 14. The apparatus of Claim 1, wherein said one or more targeted cells comprises T-cells. 15. The apparatus of Claim 1, wherein said one or more targeted cells comprises teratoma-forming cells. 16. The apparatus of Claim 1 , wherein said one or more targeted cells comprises non-tumor cells. 17. The apparatus of Claim 1, wherein said image capture system comprises a camera. 18. The apparatus of Claim 17, wherein said camera comprises a charge-coupled device. 19. The apparatus of Claim 1 , wherein said energy source comprises a laser. 20. The method of Claim 19, wherein the wavelength of energy emitted from said laser is approximately between 100 nanometers and 30 micrometers. 21. The apparatus of Claim 1, wherein said first commands comprise commands for determining the two-dimensional locations of said one or more targeted cells within said biological specimen. 22. The apparatus of Claim 1, wherein said first commands comprise commands for determining the three-dimensional locations of said one or more targeted cells within said biological specimen. 23. The apparatus of Claim 1, further comprising one or more energy beam steering devices that are controlled by said second commands. 24. The apparatus of Claim 23, wherein said energy beam steering devices comprise one or more elements that reflect the energy beam. 25. The apparatus of Claim 23, wherein said energy beam steering devices comprise one or more elements that refract the energy beam. 26. The apparatus of Claim 23, wherein said energy beam steering devices comprise one or more elements that diffract the energy beam. 27. The apparatus of Claim 23, wherein said energy beam steering devices comprise one or more elements that diffract in combination with one or more elements that refract or reflect the energy beam. 28. The apparatus of Claim 23. wherein said energy beam steering devices comprise one or more elements that refract in combination with one or more elements that reflect the energy beam. 29. The apparatus of Claim 23. wherein said energy beam steering devices comprise one or more elements that refract in combination with one or more elements that reflect and one or more elements that diffract the energy beam. 30. The apparatus of Claim 23. wherein said energy beam steering devices comprise galvanometeric mirrors. 31. The apparatus of Claim 23. wherein said energy beam steering devices comprises a piezo-electric tilt platform. 32. The apparatus of Claim 23, wherein said energy beam steering devices comprise acousto-optical deflectors. 33. The apparatus of Claim 1, wherein said first commands are executed under the control of a computer. 34. The apparatus of Claim 1 , wherein said second commands are executed under the control of a computer. 35. The apparatus of Claim 1, wherein said apparatus processes at least 1, 2, 3, 4, 5, 6, 7, or 14 square centimeters of biological specimen area per minute. 36. The apparatus of Claim 1, wherein said apparatus images at least 0.25, 0.5, 1, 2, 3, 4 or 8 million cells of said biological specimen per minute. 37. The apparatus of Claim 1. wherein said apparatus can induce a response in at least 50, 100, 150, 200, 250, 300, 350, 400, or 800 targeted cells per second. 38. The apparatus of Claim 1, wherein said response induced in said one or more targeted cells is cell death. 39. The apparatus of Claim 1, wherein said response is non lethal. 40. The apparatus of Claim 1 , wherein said response induced in said one or more targeted cells comprises activation of a photosensitive agent. 41. The apparatus of Claim 1, wherein said response induced in said one or more targeted cells comprises optoporation to allow entry of a substance. 42. The apparatus of Claim 41 , wherein said substance is genetic material. 43. The apparatus of Claim 1, wherein said response induced in said one or more targeted cells comprises inactivation of a cell component. 44. The apparatus of Claim 1 , wherein said response induced in said one or more targeted cells comprises controlled movement of said one or more targeted cells. 45. The apparatus of Claim 1, wherein said response is excitation of fluorescent dye within said one or more targeted cells. 46. The apparatus of Claim 1, further comprising an energy absorbing dye in contact with said one or more targeted cells. 47. The apparatus of Claim 1 , wherein said response is photobleaching recovery. 48. The apparatus of Claim 1 , wherein said response is to the effects of internal molecular uncaging. 49. The apparatus of Claim 1, further comprising third commands that control an autofocus mechanism linked to said image capture system. 50. The apparatus of Claim 49, wherein said third commands are executed under the control of a computer. 51. An apparatus for selectively inducing a response in one or more targeted cells within a biological specimen, comprising: a scanning lens which focuses on a field-of-view within said biological specimen, said field-of-view comprising a plurality of frames of cells; an illumination source that illuminates one or more frames of cells; an image capture system that captures an image of a first frame of cells in said field-of-view; first commands for determining the locations of said one or more targeted cells in said first frame of cells; an energy source that emits an energy beam sufficient to induce a response in at least one of said one or more targeted cells; and second commands for steering said energy beam to said locations of said one or more targeted cells within said first frame of cells. 52. The apparatus of Claim 51 , further comprising: means for directing the output of said illumination source to a second frame of cells in said field-of-view; means for capturing an image of said second frame of cells; third commands for determining the locations of said one or more targeted cells in said second frame of cells; and fourth commands for steering said energy beam to said locations of said one or more targeted cells within said second frame of cells. 53. The apparatus of Claim 52, wherein said means for directing said illumination source is one or more reflective elements. 54. The apparatus of Claim 52, wherein said means for directing said illumination source is one or more refractive elements. 55. The apparatus of Claim 51, further comprising means for moving the position of said biological specimen with respect to said scanning lens. 56. The apparatus of Claim 51, wherein said scanning lens comprises one or more refractive elements. 57. The apparatus of Claim 51, wherein said scanning lens is of F-theta design with a diameter of at least 8, 10, 12, 14, 16, or 18 mm. 58. The apparatus of Claim 51, wherein said scanning lens comprises one or more reflective elements. 59. The apparatus of Claim 51 , wherein said scanning lens comprises one or more diffractive elements. 60. The apparatus of Claim 51, wherein said scanning lens comprises one or more refractive elements in combination with one or more reflective elements. 61. The apparatus of Claim 51, wherein said scanning lens comprises one or more diffractive elements in combination with one or more refractive or reflective elements. 62. The apparatus of Claim 51, wherein said scanning lens comprises one or more refractive elements in combination with one or more reflective elements and one or more diffractive elements. 63. The apparatus of Claim 51, further comprising a proximeter to adjust the focus of the scanning lens. 64. The apparatus of Claim 63, wherein the proximeter is a laser proximeter. 65. The apparatus of Claim 51 , wherein said response is selected from the group consisting of cell death, optoporation, activation of photosensitive agent, inactivation of a cell component, controlled movement, excitation of a fluorescent reagent, photobleaching and internal molecular uncaging. 66. The apparatus of Claim 51, wherein said response is non-lethal. 67. The apparatus of Claim 66, wherein said response is selected from the group consisting of optoporation, activation of photosensitive agent, inactivation of a cell component, controlled movement, excitation of a fluorescent reagent, photobleaching and internal molecular uncaging. 68. An apparatus for identifying the cells of a first population of labeled cells within a mixture of said first population and a second population of non-labeled cells, comprising: an illumination source for illuminating a frame of cells in said mixture; an image capture system that captures an image of said frame of cells; and first commands for determining the locations of at least one cell in said first population of labeled cells by reference to said image. 69. The apparatus of Claim 68, further comprising a memory for storing said locations of said at least one cell in said first population of labeled cells. 70. The apparatus of Claim 68, further comprising a memory for storing the quantity of cells in said first population of labeled cells. 71. The apparatus of Claim 68, further comprising an energy source that emits an energy beam sufficient to induce a response in at least one cell in said first population of labeled cells. 72. An apparatus for determining a morphological or physiological characteristic of individual cells in a biological specimen, comprising: an illumination source for illuminating a frame of cells in said biological specimen; an image capture system that captures ε.n image of said frame of cells; first commands for determining the location of a first individual cell in said biological specimen by reference to said image; an energy source that emits an energy beam sufficient to induce a response in at least one of said one or more individual cells; second commands for intersecting said first individual cell with an energy beam sufficient to interrogate said first individual cell for the presence of a morphological or physiological characteristic; and a detector for measuring the response of said first individual cell to said interrogation. 73. The apparatus of Claim 72, wherein said first individual cell is identified by the presence or absence of a first label. 74. The apparatus of Claim 73, wherein said first label is a fluorescently-tagged antibody. 75. The apparatus of Claim 73, wherein said first individual cell is labeled with a second label, wherein the first label is activated by said illumination source and the second label is activated by said energy beam. 76. The apparatus of Claim 72, wherein said morphological or physiological characteristic is calcium flux. 77. The apparatus of Claim 72, wherein said morphological or physiological characteristic is cell cycle status. 78. The apparatus of Claim 72, wherein said morphological or physiological characteristic is cell mitotic history. 79. The apparatus of Claim 72, wherein said morphological or physiological characteristic is viability. 80. The apparatus of Claim 72, wherein said morphological or physiological characteristic is membrane integrity. 81. The apparatus of Claim 72, wherein said morphological or physiological characteristic is intracellular pH. 82. The apparatus of Claim 72, wherein said morphological or physiological characteristic is transmembrane potential. 83. The apparatus of Claim 72, wherein said morphological or physiological characteristic is glutathione reductive stage. 84. The apparatus of Claim 72, wherein said morphological or physiological characteristic is thiol activity. 85. The apparatus of Claim 72, wherein said morphological or physiological characteristic is expression of a gene. 86. The apparatus of Claim 72, wherein said morphological or physiological characteristic is photobleaching recovery. 87. The apparatus of Claim 72, wherein said morphological or physiological characteristic is the response to internal molecular uncaging. 88. The apparatus of Claim 72, wherein said response is non-lethal. 89. An apparatus for selectively inducing a response in one or more targeted cells within a biological specimen, comprising: an illumination source for illuminating a frame of cells in said biological specimen; an image capture system that captures an image of said frame of cells; first commands for determining the locations of said one or more targeted cells within said image; an energy source that emits an energy beam sufficient to induce a response in at least one of said one or more targeted cells within said frame of cells; and a sensor that detects the amount of energy emitted by said energy source. 90. The method of Claim 89, wherein said response is selected from the group consisting of cell death, optoporation, activation of a photosensitive agent, inactivation of a cell component, controlled movement, excitation of a fluorescent reagent, photobleaching and internal molecular uncaging. 91. The apparatus of Claim 89, wherein said response is non-lethal. 92. The apparatus of Claim 91, wherein said response is selected from the group consisting of optoporation, activation of a photosensitive agent, inactivation of a cell component, controlled movement, excitation of a fluorescent reagent, photobleaching and internal molecular uncaging. 93. A method for selectively inducing a response in one or more targeted cells within a biological specimen, comprising: illuminating a frame of cells in said biological specimen; capturing an image of said frame of cells; determining the locations of said one or more targeted cells within said image; and steering an energy beam to the locations of said one or more targeted cells, wherein said energy beam is sufficient to induce a response in at least one of said one or more targeted cells. 94. The method of Claim 93, wherein determining the locations of said one or more targeted cells comprises determining the presence of, or absence of, a label attached to said one or more targeted cells. 95. The method of Claim 93, wherein capturing an image of said frame of cells comprises capturing an image of labeled cells. 96. The method of Claim 93, wherein illuminating said frame of cells comprises illuminating said frame of cells with an excitation laser light source. 97. The method of Claim 93, wherein said response is selected from the group consisting of cell death, optoporation, activation of a photosensitive agent, inactivation of a cell component, controlled movement, excitation of a fluorescent reagent, photobleaching and internal molecular uncaging. 98. The method of Claim 93, wherein said response is non-lethal. 99. The method of Claim 98, wherein said response is selected from the group consisting of optoporation, activation of a photosensitive agent, inactivation of a cell component, controlled movement, and excitation of a fluorescent reagent, photobleaching and internal molecular uncaging. 100. A method for rapidly inducing a response in individual cells within a cell population, said cell population being held in a container, comprising: illuminating a first frame of cells in a first field-of-view in said cell population; capturing an image of said first frame of cells within said first field-of-view; determining the locations of first individual cells within said first frame of cells by reference to said image; and steering an energy beam to the locations of said first individual cells, wherein said energy beam is sufficient to induce a response in at least one of said first individual cells. 101. The method of Claim 100, further comprising: capturing an image of a second frame of cells within said first field-of-view; determining the locations of second individual cells within said second frame of cells by reference to said image; and steering an energy beam to the locations of said second individual cells within said second frame of cells, wherein said energy beam is sufficient to induce a response in at least one of said second individual cells. 102. The method of Claim 100, further comprising: illuminating a first frame of cells in a second field-of-view in said cell population; capturing an image of said first frame of cells within said second field-of- view; determining the locations of third individual cells within said first frame of cells within said second field-of-view by reference to said image; and steering an energy beam to the locations of said third individual cells, wherein said energy beam is sufficient to induce a response in at least one of said third individual cells. 103. The method of Claim 102, wherein illuminating said first frame of cells in said second field-of-view comprises moving said container and illuminating said first frame of cells. 104. The method of Claim 102, wherein said response is selected from the group consisting of cell death, optoporation, activation of a photosensitive agent, inactivation of a cell component, controlled movement, excitation of a fluorescent reagent, photobleaching and internal molecular uncaging. 105. The method of Claim 102, wherein said response is non-lethal. 106. The method of Claim 105, wherein said response is selected from the group consisting of optoporation, activation of a photosensitive agent, inactivation of a cell component, controlled movement, excitation of a fluorescent reagent, photobleaching and internal molecular uncaging. 107. A method for selectively inducing a response in one or more targeted cells within a biological specimen, comprising: focusing a scanning lens on a field-of-view within said biological specimen, said field-of-view comprising a plurality of frames of cells; illuminating one or more frames of cells within said field-of-view; capturing an image of a first frame of cells in said field-of-view; determining the locations of said one or more targeted cells in said first frame of cells; emitting an energy beam sufficient to induce a response in at least one of said one or more targeted cells; and steering said energy beam to the locations of said one or more targeted cells within said first frame of cells. 108. The method of Claim 107, wherein said response is selected from the group consisting of cell death, optoporation, activation of a photosensitive agent, inactivation of a cell component, controlled movement, and excitation of a fluorescent reagent. 109. The method of Claim 107, wherein said response is non-lethal. 110. The method of Claim 109, wherein said response is selected from the group consisting of optoporation. activation of a photosensitive agent, inactivation of a cell component, controlled movement, excitation of a fluorescent reagent, photobleaching and internal molecular uncaging. 111. The method of Claim 107, wherein capturing an image of said first frame of cells comprises capturing an image of labeled cells. 112. The method of Claim 107, wherein said illuminating one or more frames of cells comprises illuminating said one or more frames of cells with an excitation laser light source. 1 13. A method for inducing a response in one or more targeted cells within a cell population, comprising: illuminating cells in said cell population; capturing an image of the illuminated cells; determining the locations of said one or more targeted cells within said image; emitting an energy beam sufficient to induce a response in at least one of said one or more targeted cells; and measuring the amount of energy contained in said energy beam; and steering said energy beam to the locations of said one or more targeted cells. 114. The method of Claim 113, comprising measuring the amount of energy contained in said energy beam prior to steering said energy beam. 1 15. The method of Claim 1 13, wherein said response is selected from the group consisting of cell death, optoporation, activation of a photosensitive agent, inactivation of a cell component, controlled movement, excitation of a fluorescent reagent, photobleaching and internal molecular uncaging. 1 16. The method of Claim 1 13, wherein said response is non-lethal. 117. The method of Claim 116, wherein said response is selected from the group consisting of optoporation, activation of a photosensitive agent, inactivation of a cell component, controlled movement, excitation of a fluorescent reagent, photobleaching and internal molecular uncaging. 118. An apparatus for determining a morphological or physiological characteristic of individual cells in a biological specimen, comprising: an illumination source for illuminating a frame of cells in said biological specimen; an image capture system that captures an image of said frame of cells; first commands for determining the location of an individual cell in said biological specimen by reference to said image; an energy source that emits an energy beam sufficient to induce a response in at least one of the individual cells; second commands for intersecting said first individual cell with an energy beam sufficient to cause a change of a morphological or physiological characteristic of said first individual cell; and a detector for measuring a morphological or physiological characteristic of said first individual cell. 119. The apparatus of Claim 1 18, wherein said response is non-lethal. 120. The apparatus of claim 1 19, wherein said response is selected from the group consisting of optoporation. activation of a photosensitive agent, inactivation of a cell component, controlled movement, excitation of a fluorescent reagent, photobleaching and molecular internal molecular uncaging. 121. An apparatus for determining a morphological or physiological characteristic of individual cells in a biological specimen comprising: an illumination source for illuminating a frame of cells in said biological specimen, wherein said specimen contains a quantity of caged compounds; an image capture system that captures an image of said frame of cells; first commands for intersecting one or more of said quantity of caged compounds with an energy beam sufficient to uncage said one or more of said quantity of caged compounds; an energy source that emits an energy beam sufficient to uncage one or more of said quantities of caged compounds; and second commands to monitor the progress of the change in said morphological or physiological characteristic in response to the uncaged compounds of one or more of said quantity of caged compounds. 122. An apparatus for determining a morphological or physiological characteristic of individual cells in a biological specimen, comprising: an illumination source for illuminating a frame of cells in said biological specimen; an image capture system that captures an image of said frame of cells; and first comands for determining the morphological or physiological characteristic of said individual cell in said frame of cells. 123. The apparatus of Claim 122, wherein said morphological or physiological characteristic is calcium flux. 124. The apparatus of Claim 122, wherein said morphological or physiological characteristic is cell cycle status. 125. The apparatus of Claim 122, wherein said morphological or physiological characteristic is cell mitotic history. 126. The apparatus of Claim 122, wherein said moφhological or physiological characteristic is viability. 127. The apparatus of Claim 122, wherein said moφhological or physiological characteristic is membrane integrity. 128. The apparatus of Claim 122, wherein said moφhological or physiological characteristic is intracellular pH. 129. The apparatus of Claim 122, wherein said moφhological or physiological characteristic is transmembrane potential. 130. The apparatus of Claim 122, wherein said moφhological or physiological characteristic is glutathione reductive stage. 131. The apparatus of Claim 122, wherein said morphological or physiological characteristic is thiol activity. 132. The apparatus of Claim 122, wherein said moφhological or physiological characteristic is expression of a gene. 133. The apparatus of Claim 122, wherein said moφhological or physiological characteristic is photobleaching recovery. 134. The apparatus of Claim 122, wherein said moφhological or physiological characteristic is the response to the internal molecular uncaging.

Inventors: EISFELD TIMOTHY M, KOLLER MANFRED R, O'NEAL ROBERT A, PALSSON BERNHARD O, EISFELD, TIMOTHY, M., KOLLER, MANFRED, R., O'NEAL, ROBERT, A., PALSSON, BERNHARD, O.
Applicants: ONCOSIS
Application year: 2000
Main classification: C12N13
IPC codes: C12M1, C12N13, C12N15, C12Q1, G01N33