Assembly for optical pre-treatment of optically activatable biological samples - Patents.com

Abstract

An assembly is described for optically pre-processing an optically activatable biological sample containing a plurality of cells suspended in a liquid, the assembly comprising a reservoir for storing the sample, from which the sample can be transported by a transport unit through a hollow channel, the cells being transportable one after another along the hollow channel, an illumination unit arranged along the hollow channel for illuminating the cells contained in the sample, the cells flowing through the hollow channel at a flow rate that can be determined by the transport unit as set by controllable illumination intensity and illumination duration, and further comprising a cell analysis and / or sorting device fluidly connected to the hollow channel downstream of the hollow channel.

Description

[Technical field]

[0001] The present invention relates to an assembly for the optical pre-treatment of an optically activatable biological sample. [Background technology]

[0002] For both quantitative measurement and molecular characterization of living cells, so-called flow cytometers are used, which comprise a flow measurement cell, usually in the form of a light-transmitting microchannel cell, through which isolated cells from a stored cell suspension flow continuously. A light source assembly, usually in the form of at least one laser, is arranged along the microchannel cell, from which a light beam illuminates or passes laterally through the individual cells as they pass along the microchannel cell through a specific measurement zone. By means of appropriately positioned light detectors, it is possible to detect both the scattered components of the stimulating laser light and the phenomenon of fluorescence caused by the laser beam, which usually originates from fluorescent labels attached to the cells or cell components, and which is used for the simultaneous analysis of the physical and molecular properties of the individual cells.

[0003] WO 2005 / 017498 discloses a conventional flow cytometer in which light emitting diodes, or LEDs for short, are used instead of the lasers mentioned above, and individual cells are illuminated at different angles of incidence and / or at different wavelengths. In a suitable format, multiple detectors are used to detect scattered light components as well as fluorescent emissions by the cells or emitted from fluorescent labels attached to the cells.

[0004] In order to improve the reproducibility and significance of measurements obtained by optical cell measurements using a flow cytometer, WO 2017 / 036999 describes a device for optically stimulating an optically activatable biological sample. The biological sample is stored in a sample container, which is optically and thermally coupled to a temperature-controlled fluid circuit through a hollow channel, and at least one light source is arranged on or in the hollow channel and thermally coupled thereto, which can illuminate the optically activatable biological sample stored in the sample container in a controlled manner, after which the sample is aspirated from the sample container and fed to a flow cytometer.

[0005] If cell sorting is required instead of or in combination with cell analysis, cells suspended in a translucent liquid, optionally after passing through a flow cytometer, usually enter a microcapillary through which isolated cells flow in succession one after the other. Typically, a laser beam is applied to the cells flowing along the microcapillary, and if necessary, fluorescence is stimulated if a fluorescent label is attached to the cells. The scattered light and, if necessary, fluorescence light generated by each cell is detected by a detector, and the resulting detector signal is the basis for the subsequent sorting mechanism. A transducer attached to the capillary outlet separates the liquid stream into small droplets, which pass through an electrostatic sorting mechanism after leaving the microcapillary, whereby the cells are separated and reach spatially separated collection vessels according to the sorting specifications.

[0006] This system for cell analysis and sorting constitutes a tool for experiments in the field of optogenetics, with which useful information can be obtained about intracellular and extracellular processes. In particular, optogenetics makes it possible to analyze and possibly control highly complex and above all rapid biochemical reactions, but also the transmission of bioelectrical signals in cells. However, the available means for cell analysis and sorting offer only limited possibilities for controlling the optical interaction with the living cells to be analyzed and for controlling the exertion of influence on intracellular and extracellular processes. Summary of the Invention [Problem to be solved by the invention]

[0007] Description of the Invention The aim of the present invention is to take steps that make it possible to significantly expand the scope for obtaining information and for influencing or controlling intracellular and extracellular events, compared to the above-mentioned techniques currently applied and available.

[0008] Due to the complexity of the intracellular structures and the associated biochemical and bioelectrical events occurring both within and between cells, the understanding of cells and cellular processes as a whole remains incomplete. It is currently believed that one of the reasons for this is that the cells fed into the flow cytometer are in a substantially non-specific, i.e. imprecisely defined, state. The same is true for the cell sorting process. [Means for solving the problem]

[0009] The basic idea of ​​the present invention is to controllably illuminate individual cells over time, immediately prior to a cell analysis known per se, using a flow cytometer or cell sorter, preferably based on fluorescence-based cell sorting, with a specific amount of light for a specific period of time, so that specific functional events within the cells can be optically activated or deactivated, as explained below, to obtain specific cell states that can be precisely analyzed or that offer the opportunity to perform specific manipulations or procedures on the cells.

[0010] The object of the invention is achieved as defined in claim 1. Further features which advantageously develop the inventive concept can also be taken from the following description which forms the subject matter of the dependent claims and which is given with reference to exemplary embodiments. Claim 17 defines the use of the assembly according to the invention for the positive selection of certain biological cells from a cell suspension which contains a variety of biological cells.

[0011] According to the solution, an assembly for optical pre-treatment of an optically activatable biological sample comprising a plurality of cells suspended in a liquid is provided, the assembly comprising a reservoir for storing the sample, from which the sample can be transported by a transport unit through a hollow channel, along which the cells can be transported successively, i.e. preferably individually, one after the other, along which an illumination unit is arranged for illuminating the cells contained in the sample, the cells flowing through the hollow channel at a flow rate that is set by a controllable illumination intensity and illumination duration and that can be determined by the transport unit. Downstream of the hollow channel, a cell analysis and / or sorting device is attached in fluid communication with the hollow channel.

[0012] An assembly according to the invention, which may also be configured and used as a modular illumination unit or illumination attachment fixed upstream in the flow direction of a known flow cytometer or cell sorter, will now be described in more detail with reference to the drawings.

[0013] Although the control of the illumination of the whole cell sample in relation to known flow cytometers is known for cell analysis, this known illumination form is not compatible with sorting flow cytometers, abbreviated as "FACS devices" (fluorescence activated cell sorting) (FACS is a registered trademark of Becton, Dickinson and Company). The sample chamber of these devices is small, integrated inside the respective device and is also pressurized. This makes it almost impossible to use an illumination attachment with this type of fluorescence activated cell sorting device. In addition, in this type of sorting flow cytometer, the whole cell sample is illuminated and then taken into the cytometer. This is not sufficient for cell sorting procedures, especially since the time delay occurring between illumination and the measurement / sorting of the individual cells must be identical. Of course, it is possible to envisage the illumination of the cells along the existing capillary system of the FACS equipment, but it is not possible to fully control the illumination duration, the illumination intensity or the temperature of the cell sample.

[0014] The solution described herein avoids the pre-integrated sample chamber of known FACS machines and preferably uses an external transport unit that allows control of the sample flow and thus manages the illumination and temperature control of the cell sample along the capillary. The most important aspect in this respect is that the time interval between illumination and measurement / sorting is the same for each individual cell, thus resulting in the same sorting time for each individual cell across the cell sample.

[0015] The invention will now be described by way of example with reference to the drawings using exemplary embodiments, without limiting the general inventive concept. [Brief description of the drawings]

[0016] [Figure 1] FIG. 1 shows an assembly according to the invention having a number of individual light sources. [Diagram 2] FIG. 1 shows an assembly according to the invention having at least one, and preferably several, light guides for illuminating cells. [Diagram 3] FIG. 1 shows an assembly for positively selecting specific cells from a cell suspension. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0017] Modes for carrying out the invention, industrial applicability 1 shows a block diagram of an assembly according to the invention, which comprises the following components: A reservoir 1 stores an optically activatable biological sample 2 in which a plurality of biological cells 3 are suspended in a translucent, i.e. light-transmitting, liquid 4. The temperature of the biological sample 2 in the reservoir 1 can be controlled by means of a thermal unit 5, preferably at an identifiable temperature.

[0018] By means of a fluid supply pump 6, the discharge or supply rate v of which can be controlled by a control and / or regulating device 7, the biological sample 2 stored in the reservoir 1 enters the capillary 9 via a fluid line 8. The capillary 9 comprises a hollow channel 10 with a capillary diameter 11, the dimension of which is preferably not larger than the sum of the diameters of two cells 3 contained in the sample 2. That is to say, the cells 3 flowing along the capillary 9 preferably flow through the capillary 9 one by one, i.e. one after the other in succession. Nevertheless, the dimension of the capillary diameter can also be larger, for example up to 200 μm, depending on the cells present in the suspension, and more than two cells, for example from 3 to preferably up to 20 cells, can flow along the capillary next to each other. What is important here is that the cell illumination for the individual cells is identical at a defined time point, so that each cell can be brought into a predefined optically excited state. The capillary 9 has a capillary wall 12 that is transparent to light.

[0019] In the exemplary embodiment shown in Fig. 1, outside the hollow channel 9, i.e. outside the capillary 9, a number of individual light sources 13 are arranged at least partially along the hollow channel 10, next to each other in line axially with respect to the hollow channel, and can be controlled individually or in groups (131, 132, 133) by a control and / or regulating device 7. Preferred light sources are individual light emitters or hybrid light emitters such as LEDs, laser diodes, halogen lamps, gas discharge lamps, LCD, LED or OLED display units, projectors or quantum dots.

[0020] The individual light sources 13 are preferably arranged next to each other not only in the axial direction but also in the circumferential direction of the hollow channel 10. In this way, the cells 3 flowing in the hollow channel 10 near the light sources 13 are uniformly illuminated from all sides.

[0021] In order to make the light input to the individual cells 3 as they pass through the hollow channel 10 or capillary 9 as individual and diverse as possible, the individual light sources 13 are collected into groups 131, 132, 133. The light sources of each associated group 131, 132, 133 each emit a specific wavelength λ of specifically definable light intensity. 131 , λ 132 , λ 133 It emits light with wavelength λ 131 , λ 132 , λ 133 and the associated light intensities are preferably different from one another. In principle, the individual light source groups 131, 132, 133 etc. can be selected such that each light source group 131, 132, 133 etc. comprises at least one light source 13.

[0022] A large number of light sources 13 arranged not only axially but also circumferentially around the hollow channel 10 allows the light input to each individual cell 3 to be individually determined in terms of irradiance or intensity, and also in terms of wavelength, using the control and adjustment unit 7.

[0023] The optically pre-treated cells 3 leaving the capillary 9 enter, via a further fluid line 14, a cell analysis and / or sorting device 15 known per se.

[0024] Advantageously, the capillary 9 is thermally coupled to a heat exchanger 16 that allows a determinable temperature to be obtained for the biological sample 2 in the capillary 9. The heat exchanger 16 may be a Peltier element or, as can be seen in FIG. 1, is configured in the form of a temperature control unit T connected to a fluid circuit 17 along which a heat transfer fluid is supplied, at least part of which passes through an annular channel 18 defined radially outwardly by a hollow cylinder H that radially surrounds the hollow channel 10 or the capillary 9. The heat transfer fluid flowing through the annular channel 18 is thermally coupled to the hollow channel wall or capillary wall associated with the hollow channel, making it possible to control the temperature of the biological sample 2 flowing in the hollow channel 10. Alternatively or in combination, the temperature control unit T may be coupled to the heat exchanger, for example in the form of an air / liquid heat exchanger or by means of a so-called heat pipe.

[0025] In a further preferred embodiment, the individual light sources 13 are at least partially arranged in annular channel 18, whereby a heat transfer fluid can flow around the individual light sources 13 and maintain the individual light sources 13 at a uniform, identifiable temperature level, thus preventing local overheating that may be caused by the individual light sources 13.

[0026] Due to the determineable flow rate v obtained by means of the feed pump 6, the assembly according to the invention makes it possible to precisely determine the residence time and thus the illumination period for individual cells 3 in the capillary 9. Due to the plurality of individual light sources 13, which can be operated individually or in groups in a wavelength-selective and radiation intensity-controlled manner by means of the control and regulation unit 7, the amount or intensity of light and the wavelength or spectrum of wavelengths of light applied to the individual cells 3 can be determined individually. Thus, the biological cells 3 can be optically treated in an determineable manner, for example immediately before a cell analysis known per se with a flow cytometer or before cell sorting.

[0027] In Fig. 2 an alternative embodiment for carrying out an assembly according to the invention for optically pre-treating an optically activatable biological sample 2 is shown. In contrast to the assembly shown in Fig. 1, which envisages a number of individual light sources 13 for the purpose of illuminating or controlling the irradiation of the cells 3 flowing through the capillary 9, the embodiment according to Fig. 2 has at least one light guide 19, for example in the form of a glass fibre, arranged outside the hollow channel 10 along the capillary 9. The light guide 19 is connected to a light source 20. In addition, the light guide 19 is provided with at least one light exit zone 21, transversely to its longitudinal extension, directed towards the hollow channel 10, through which light can enter the hollow channel 10. The at least one light exit zone 21 can for example be produced by local roughening of the light guide 19. The roughening allows scattered light components to exit laterally from the light guide 19. The axial length l of the light exit zone 21 allows the illumination or illumination period of the cells 3 passing through the capillary 9 at a particular flow rate to be determined.

[0028] The light guide 19 shown in FIG. 2 has a total of three light exit zones 21 which are axially separated from one another and have the same dimensions.

[0029] Optionally, at least one second light guide 20 may be disposed along the capillary 9, into which light from a light source 22 is also coupled. The light sources 20 and 22 may be identical or may provide different wavelengths. In addition, the number, arrangement and length of the light exit zones 23 along the light guide 20 may differ from the light exit zones 21 of the light guide 19. Obviously, light guides of this type may be disposed in almost any number along and around the capillary 9.

[0030] 2 also has a heat exchanger 16 with an associated fluid circuit 17 passing through an annular channel 18 arranged at least in part alongside the capillary 9. Optionally, light guides 19, 20 are arranged within the annular channel 18 through which the heat transfer fluid flows, thus providing uniform temperature control.

[0031] By the controlled optical exposure according to the invention of the biological cells 3 flowing one after the other in succession through the capillary 9, and optionally by means of dyes or fluorescent substances or light-modulating particles attached or bound to the cells 3, the cells are transformed into a defined state which forms the basis for reproducible cell analysis, cell manipulation and / or cell sorting. Due to the temporally and spatially defined sequence of optical cell illumination and subsequent cell analysis and / or cell sorting, precise optogenetic experiments and procedures can be performed. Instead of cell analysis using a flow cytometer, the use of analytical devices, for example mass spectrometers or magnetic purification systems using magnetic beads etc. is also possible.

[0032] In this way, with this device it is possible to verify the so-called Kinetic Proof Reading Model (KPR), which shows that T cells distinguish between endogenous and exogenous ligands by differences in the half-life of the ligand binding to the T cell receptor (TCR). For this reason, the plant photoreceptor phytochrome B can be used to selectively investigate the kinetics of ligand binding to the TCR by controlling the illumination. The reproducible optical pre-treatment of the investigated T cells, which can be obtained with the device according to the invention, allows scientifically relevant measurements for the determination of the half-life of the ligand-TCR interaction, which is a decisive factor for the activation of downstream TCR signaling.

[0033] A further preferred embodiment of the assembly according to the invention is shown in Fig. 3, which makes it possible to carry out a positive selection of cells from a mixture of cells in the form of a cell suspension, as is the case for example with blood. This possibility of positive cell selection is particularly advantageous compared to conventional methods in tumor research and cancer diagnosis.

[0034] In particular, immune cells are activated by the binding of antibodies to their surface receptors, and die as a result of activation.This is why, mainly, immune cell subtypes can only be negatively selected, that is, a cocktail of antibodies is required and must be made first, which labels all cells except the target cells to be selected.This procedure is very costly in terms of time, procedures and techniques, and rarely leads to the isolation of a truly pure cell population.

[0035] Using the assembly and procedure shown in FIG. 3, cells to be positively selected are bound to the light-modulating particles for a very short period of time without being activated thereby, with no lethal consequences.

[0036] A cell suspension, for example in the form of a blood sample containing various cells 3, for example immune cells, so-called T-cells, is present in a reservoir 1 configured similarly to the reservoir of figures 1 and 2. The cells 3 are mixed with optically activatable particles 24, for example in the form of photo-controllable binding molecules, photo-controllable antibodies, photo-controllable single domain antibodies (nanobodies) or photo-controllable adnectins (monobodies).

[0037] The optically activatable particles 24 are transformed from a first particle state into specific cells 3 of the cell suspension to be separated by a first optical activation in the form of a conformational change. * A second optical activation and associated required second conformational change causes the optically activated particle to resume an unbound state and bind to a specific cell 3. * The particle can then transition again to the first particle state, which can then be released from the

[0038] The cell suspension stored in the reservoir 1 is transferred by the transport unit 6 along the capillary 9 into an assembly 25 for optical pre-treatment.

[0039] The transport unit 6 transports the cell suspension at a specifiable flow rate, as set by a controllable illumination intensity and duration, through a hollow channel or capillary 9, along which the cell suspension is illuminated by an illumination unit 26 arranged in an assembly 25 for optical pre-treatment. This first optical activation causes the optically activatable particles 24 to adopt a second particle state and to become specific cells 3. * All remaining cells 3' in the cell suspension retain their original morphology.

[0040] Inside the sorting unit 15 downstream of the optical pre-treatment assembly 25, the optically activated cell suspension is subjected to optically and / or magnetically induced sorting to separate the cells 3 contained in the cell suspension. * , 3' are sorted and separated, preferably based on the amount of fluorescence emitted and / or the spectral color analysis and / or magnetic properties. In this way, the specific cells 3 to which the optically activated particles 24 are bound are identified. * The optically activated particles 24 are preferably coupled to a dye, and therefore may emit more fluorescence than all other cells 3'. Alternatively, the use of optically activatable particles 24 coupled to magnetic particles, so-called magnetobeads, may be considered. In this case, the specific cells 3' to which the magnetobeads are coupled via the optically activated particles may be detected. * can be separated by the sorting unit 15 based on magnetic force.

[0041] Downstream of the sorting unit 15 are at least two sample collection vessels 27, 28. The optically activated particles 24 are bound to the specific cells 3 to be positively selected. * All of the above enters the sample collection container 28. A further illumination unit 29, which is arranged between the sorting unit 15 and the sample collection container 28 or in or on the sample collection container 28, serves for a second optical activation, whereby specific cells 3 * The particle 24 bound to the specific cell 3 then transitions back to the first particle state through a conformational change. *All remaining cells 3' enter another sample collection vessel.

[0042] Subsequent separation 30 isolates the optically activatable particles 24 from specific cells 3, e.g., using centrifugation, decanting, filtration, magnetic separation, etc. * As a result, specific cells are isolated from the * A pure population of 31 is obtained. [Explanation of symbols]

[0043] 1 Reservoir 2. Biological samples 3 cells 3 * Specific cells 3' Remaining cells 4. Translucent liquid 5 Thermal Unit 6 Fluid supply pump 7 Control and regulating devices 8 Fluid Lines 9 Capillary 10 Hollow Channel 11 Capillary diameter 12 Capillary Wall 13 Light source 131~133 Light source group 14 Fluid Lines 15 Cell analysis or cell sorting devices 16 Heat Exchange Unit 17 Fluid circuit 18 Annular Channel 19 Light Guide 20 light source 21 Light Exit Zone 22 Light source 23 Light Exit Zone 24 Optically Activatable Particles 25 Optical Pre-Processing Assembly 26 Lighting Unit 27 Sample collection container 28 Sample collection container 29 Further Lighting Units 30 separation 31 Pure populations of specific cells l Length of the optical exit zone T Temperature Control Unit H Hollow cylinder v Feed rate

Claims

1. 1. An assembly for optically pre-processing an optically activatable biological sample comprising a plurality of cells suspended in a liquid, the assembly comprising a reservoir for storing said sample, The assembly, characterized in that the sample can be transported through a hollow channel by a transport unit outside the reservoir, the hollow channel is dimensioned to transport the cells along the hollow channel one after the other and up to 20 of the cells next to each other, an illumination unit is arranged along the hollow channel to illuminate the cells contained in the sample, the cells flow through the hollow channel at a flow rate that is set by a controllable illumination intensity and illumination duration and can be determined by the transport unit, and further comprising a cell analysis and / or sorting device in fluid communication with the hollow channel downstream of the hollow channel.

2. 2. The assembly described in claim 1, characterized in that the hollow channel is configured in the form of a capillary that is transparent to light, the capillary diameter having a dimension not greater than the sum of the two diameters of the cells contained in the sample.

3. 3. The assembly according to claim 1 or 2, characterized in that the lighting unit comprises a first plurality of individual light sources arranged outside the hollow channel, the first plurality of individual light sources being arranged next to each other in an axial direction relative to the hollow channel, at least partially along the hollow channel, and can be controlled individually or in groups.

4. 4. The assembly according to claim 3, characterized in that the lighting unit comprises at least one second plurality of individual light sources arranged outside the hollow channel, the second plurality of individual light sources being offset at least in the circumferential direction of the hollow channel with respect to at least the first plurality of light sources, arranged at least partially along the hollow channel and axially aligned with respect to the hollow channel, and can be controlled individually or in groups.

5. 5. The assembly according to claim 3 or 4, characterized in that the light sources consist of individual light emitters or hybrid light emitters of LEDs, laser diodes, halogen lamps, gas discharge lamps, LCD, LED or OLED display units, projectors or quantum dots.

6. The lighting unit comprises at least one light guide arranged along the hollow channel, the light guide having at least one light exit zone directed towards the hollow channel transversely to the longitudinal extension of the light guide, The light guide is optically coupled to a light source for coupling light into the light guide.

6. An assembly according to any one of claims 1 to 5, characterized in that

7. 7. An assembly according to claim 1, wherein the hollow channel is thermally coupled to a heat exchanger.

8. 8. The assembly according to claim 7, characterized in that the heat exchanger is configured in the form of a hollow cylinder radially surrounding the hollow channel and encloses an annular channel with a hollow channel wall associated with the hollow channel, through which flows a temperature-controlled liquid that is thermally coupled to the hollow channel wall and is also thermally coupled to the sample in the hollow channel.

9. 9. The assembly of claim 8, wherein at least a portion of the lighting unit is disposed within the annular channel and is thermally coupled to the temperature-controlled liquid.

10. 10. The assembly according to claim 1, characterized in that a control and / or adjustment device is provided for controlling or adjusting the transport unit and / or the illumination unit according to a time-defined, identifiable period of time so as to irradiate the cells passing successively through the hollow channel with light of an identifiable constant light intensity and of wavelengths of an identifiable wavelength spectrum.

11. Assembly according to any one of claims 7 to 9 or claim 10 relying on claims 7 to 9, characterized in that the control and / or regulating device monitors the heat exchanger in order to control an identifiable temperature.

12. the cell analysis device is a flow cytometer; The cell sorting device is a fluorescence activated cell sorter Assembly according to any one of claims 1 to 11, characterized in that

13. 13. The assembly of claim 12, wherein the flow cytometer comprises at least one pressure source that functions as a drive source for the transport unit.

14. Assembly according to any one of claims 1 to 13, characterized in that the conveying unit is a liquid pump in the form of a membrane pump or a syringe pump.

15. the cell sorting device includes a sorting mechanism having at least two downstream sample collection vessels for respectively receiving components of the sample; At least one further illumination unit is provided for illuminating the sample collected in at least one sample collection vessel. Assembly according to any one of claims 1 to 14, characterized in that

16. 16. The assembly according to claim 15, characterized in that the at least one further lighting unit is arranged between the sorting mechanism and the sample collection vessel, within the sample collection vessel or on the sample collection vessel.

17. 15. Use of the assembly according to any one of claims 1 to 14 for positively selecting a specific biological cell from a cell suspension containing different biological cells, wherein the cell sorting device has a sorting mechanism with at least two downstream sample collection vessels for respectively receiving components of the sample, and at least one further illumination unit is provided for illuminating the sample collected in at least one of the sample collection vessels.

18. 18. Use of an assembly according to claim 17, characterized in that the at least one further lighting unit is arranged between the sorting mechanism and the sample collection vessel, within the sample collection vessel or on the sample collection vessel.

19. The cell suspension is stored in a reservoir as a sample together with optically activatable particles, which can be transferred by a first optical activation in the form of a conformational change from a first particle state to a second particle state in which the optically activated particles bind to specific cells of the cell suspension, and can be transferred by a second optical activation in the form of a second conformational change back to the first particle state in which the optically activatable particles adopt a non-bound state and are released from the specific cells, the sample is transported from the reservoir by the transport unit through the hollow channel at a specifiable flow rate, along which the sample is illuminated by the illumination unit with a controllable illumination intensity and duration set so that the optically activatable particles adopt the second particle state and bind to the specific cells; a sorting device disposed sequentially along the hollow channel separating the specific cells having at least one optically activated particle bound thereto into a sample collection vessel; A further illumination unit disposed downstream of the sorting device causes the particles bound to the specific cells to be reverted to the first particle state by the second optical activation and released from the specific cells. Use according to claim 17 or claim 18, characterized in that

20. 20. The use according to claim 19, characterized in that the optically activatable particles are in the form of photocontrollable binding molecules, photocontrollable antibodies, photocontrollable single domain antibodies (nanobodies) or photocontrollable adnectins (monobodies).

21. 21. Use according to claim 19 or claim 20, characterised in that the optically activatable particles are distinguished by at least one characteristic colour and / or by fluorescent properties and / or by magnetic properties.

22. Use according to any one of claims 17 to 21, characterized in that the cell suspension is blood and the specific cells are immune cells, T cells.

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