Automation of incubation, processing, harvesting and analysis of samples in a multi-cell plate with thin film sample support

Abstract

The present invention relates to the automation of incubation, processing, harvesting and analysis of samples in a multi-cell plate. In particular, a multi-cell plate including a body with a plurality of cells is presented. Furthermore, an automated crystal harvesting and processing system with a cutting unit, a fluid unit and a removing device is presented. The multi-cell plate further includes a sealing film for sealing the cells on a first side of the body and a sample film for sealing the cells on a second side of the body. The sample film is adapted for accommodating a biological material for crystallization. Furthermore, the sample film is of a thickness and composition that makes it compatible with x-rays and also with laser ablation. The design of the multi-cell plate and the automated crystal harvesting and processing system allows for several steps of incubation, processing, harvesting and analysis of the samples to be automated.

Claims

1. Multi-cell plate for automated incubation, processing harvesting and analysis of samples of biological material, the multi-cell plate comprising a body with a plurality of cells; a sealing film for sealing the cells on a first side of the body; a sample film for sealing the cells on a second side of the body and for accommodating a biological material for crystallization; wherein the sample film comprises a cyclic olefin copolymer material and comprises a thickness between 1 μm and 50 μm.

2. Multi-cell plate according to claim 1, wherein the sample film comprises an anti-adhesive material.

3. Multi-cell plate according to claim 1, wherein the body comprises a predetermined pattern of markers, wherein the markers are selected from the following group of markers: recess, protuberance and optical marker.

4. Automated system for incubation, processing, harvesting and analysis of samples of biological material, the system comprising a multi-cell plate according to claim 1; a cutting device for penetrating the sample film.

5. Automated system according to claim 4, wherein the cutting device penetrates the sample film by photo ablation.

6. Automated system according to claim 4, further comprising an identification device for automatically identifying a cell and/or a position of a crystal in a cell of the multi-cell plate based on markers on the multi-cell plate and/or based on image analysis.

7. Automated system according to claim 4, wherein the cutting device is one of a nano second laser, a pico second laser and a femto second laser.

8. Automated system according to claim 4, further comprising a fluid unit for supplying a fluid to a cell and/or to a crystal in the cell of the multi-cell plate and/or for extracting a fluid from a cell of the multi-cell plate; wherein the cutting device provides an opening in the sample film; wherein the fluid unit is connectable via the opening to the cell of the multi-cell plate.

9. Automated system according to claim 8, wherein the cutting device varies the dimensions and/or shape of the opening automatically depending on the amount of the fluid to be supplied or extracted.

10. Automated system according to claim 8, wherein the fluid unit supplies at least one of a cryo protectant and a ligand solution to a cell of the multi-cell plate.

11. Automated system according to claim 8, wherein the fluid unit applies a negative pressure to a cell of the multi-cell plate.

12. Automated system according to claim 4, further comprising a removing device for removing the sample from the multi-cell plate by applying negative pressure to the sample film; wherein the cutting device cuts around a sample on the sample film.

13. Method for operating the automated system according to claim 4, the method comprising the following steps identifying a cell and/or a position of crystals in a cell of the multi-cell plate; providing an opening in the sample film of the identified cell by a cutting device; connecting a fluid unit to the cell via the opening; supplying a fluid to the cell or extracting a fluid from the cell by the fluid unit.

14. Method according to claim 13, further comprising wherein the identifying takes place automatically based on markers on the multi-cell plate and/or based on image analysis by an identification device.

15. Method according to claim 13, wherein a fluid is supplied to the cell, the fluid influencing a crystallization process of a crystal comprised within the cell.

Description

BRIEF DESCRIPTION OF THE DRAWINGS

(1) Exemplary embodiments of the invention will be described in the following with reference to the following drawings.

(2) FIG. 1 shows a perspective view of a multi-cell plate according to an embodiment of the invention

(3) FIG. 2 shows a schematic representation of an automated system for incubation, processing, harvesting and analysis of samples of biological material according to a further embodiment of the invention

(4) FIG. 3 shows a shows a schematic representation of an automated system according to a further embodiment of the invention

(5) FIG. 4 shows a flow diagram of a method for processing the multi-cell plate according to embodiment of the invention

(6) FIG. 5 shows a flow diagram of a method for processing the multi-cell plate according to a further embodiment of the invention

(7) FIG. 6 shows a flow diagram of a method for automated incubation, processing, harvesting and analysis of samples of biological material according to an embodiment of the invention

(8) FIG. 7 shows part of a flow diagram of a method for automated incubation, processing, harvesting and analysis of samples of biological material according to a further embodiment of the invention

(9) FIG. 8a/b show examples of a removing device applicable for a multi cell plate according to an embodiment of the invention

DETAILED DESCRIPTION OF EMBODIMENTS

(10) In FIG. 1 a multi-cell plate 1 is shown in a perspective view. The multi-cell plate 1 is shown in a sitting drop application in which a sample 15 of biological material is incubated on a sample film 11 and the multi-cell plate 1 is oriented in such a way that during crystal growth the sample film 11 is located in the lower part of the multi-cell plate 1. Alternatively, the multi-cell plate 1 may be used in a hanging drop application (not shown in the Figures). In the hanging drop application the multi-cell plate 1 is oriented in such a way that the sample 15 and thus the sample film 11 is located in the upper part of the multi-cell plate 1, i.e. above the sealing film 7.

(11) The multi-cell plate 1 comprises a body 3 with a plurality of cells 5. In the example shown, in each of the cells 5 a precipitate reservoir 19 with a precipitate liquid is located. The precipitate liquid induces a formation of crystals from the sample drop on the sample film 11 by vapor diffusion. The cells 5 are sealed by a sealing film 7 on a first side 9 of the body 3. Furthermore, on the opposing second side 13 of the body 3 the cells 5 are sealed by a sample film 11. The cells may be sealed airtight by the sealing film 7 and by the sample film 11.

(12) The sample film 11 is made of a material compatible with photo ablation. Preferably, the sample film 11 is made of a cyclic olefin copolymer. Furthermore, the sample film 11 may include further polymers such as polyimide. Moreover, the sample film 11 is preferably cutable by laser ablation. The sample film 11 may be coated with a material, such as black carbon, which aids the cutting of the sample film 11. Furthermore, the sample film 30 preferably has a thickness between 1 and 50 μm.

(13) The material and the thickness of the sample film 11 are designed to provide a minimum of X-Ray scattering during an analysis of the sample 15. In addition, the sample film 11 may be coated with an anti-adhesion material such as PTFE for preventing the sample from adhering to the sample film 11. Furthermore, the sample film 11 may be inert to the sample 15, to the sample solution and to the precipitate liquid.

(14) By employing a sample film 11 consisting of cyclic olefin copolymer for the multi-cell plate 1 several steps of processing, harvesting and analysis of samples 15 may be automated and manual interaction with the samples may be avoided. This is due to the properties of the cyclic olefin copolymer material such a compatibility with photo ablation which allows an automation of a harvesting process. Furthermore, the cyclic olefin copolymer allows a small opening to be made in the film by laser ablation. The opening may be used for automatically supplying cryo protectants or ligands to the sample or for removing the sample solution as described below. Moreover, the high transparency and low birefringence of the cyclic olefin copolymer allow an in-plate-data-collection which reduces operator interactions with the sample.

(15) Furthermore, for reducing user interactions the multi-cell plate 1 comprises markers 17 designed e.g. as holes, pins, recesses and optical patterns. The markers 17 are preferably arranged at the body 3 of the multi-cell plate 1 and may be used as holding devices for a robotic arm or for a positioning unit such as a plate support. Based on the locations of the markers the orientation and location of certain cells 5, and particularly, the orientation and location of samples 15, i.e. crystals, in the cells 5 of the multi-cell plate may be determined thus providing further automation of the incubation, processing, harvesting and analyzing of the samples.

(16) In FIG. 2 a schematic view of an automated system 21 for incubation, processing, harvesting and analysis of samples 15 of biological material is presented. The functionality of the system is explained in greater detail with respect to FIGS. 4 to 6.

(17) The automated system 21 comprises a cutting device 23 designed as a mechanical cutter or preferably as a femto second laser. The cutting device 23 may include a scanning optic device e.g. for focusing and directing the beam of the laser onto a selected cell 5. Moreover, the system 21 comprises a positioning device 25. The positioning device 25 may comprise a robotic arm 35 e.g. for transferring the multi-cell plate 1 between different units of the system 21. Furthermore, the positioning device 25 may comprise one or several plate supports 37. The plate support may be e.g. an X-Y-platform movable by drives for positioning and orientating the multi-cell plate 1 within one of the units of the system 21. The positioning device 25 may be employed for receiving the multi-cell plate 1 and for aligning the plurality of cells 5 for rapid and automatic identification of the cells 5 e.g. initially by using the information on the geometry of the plate and more precisely by an identification device 27 and for automatically aligning the crystals under the working area of the laser. For this purpose, crystal positions recorded relative to the reference markers 17 may be used.

(18) Moreover, the system 21 may comprise an identification device 27 for automatically identifying a cell 5 of the multi-cell plate 1 based on markers 17 on the multi-cell plate 1 and/or based on image analysis. The identification device 27 may comprise a microscope or a camera and may be connected to a control device 39 on which the predetermined positions of the markers 17 or an image analysis program are stored.

(19) The automated system 21 may further comprise a fluid unit 29 for supplying a fluid to a cell 5 of the multi-cell plate 1 and/or for extracting a fluid from a cell 5 of the multi-cell plate 1. Therein, the fluid unit 29 may be divided into a solution delivery unit for supplying chemicals to the samples 15 and into an aspiration unit for withdrawing fluid from the cells 5. The fluid unit 29 as well as the cutting device 23 and the positioning device 25 are connected to the control device 39.

(20) The control device 39 is adapted to initiate a cutting of a hole by the cutting device 23 in the sample film 11 of the multi-cell plate 1. Furthermore, the fluid unit 29 is connectable via the opening to a selected cell 5 of the multi-cell plate 1. The operation of the automated system 21 is described in detail below.

(21) As shown in a further example in FIG. 3 the automated system may comprise further units and devices e.g. also connected to the control device 39. For example, the system 21 further comprises a removing device 33 also denoted as harvesting device. The removing device 33 may be designed as a hollow pin and may be adapted for applying negative pressure to the sample film 11 of the multi-cell plate 1. For this purpose the removing device 33 may be connected to the fluid unit 29. Therein, the removing device 33 may remove a sample 15 together with a part of the sample film 11 after the cutting device 23 cuts around the sample 15. The removing device 33 may be movable by the robotic arm 35.

(22) FIGS. 8(a) and (b) show examples of removing devices 33 provided as hollow pins 34. In FIG. 8(a), the removing device 33 comprises a beveled tip with a plane slanted surface 53 adjacent to an end opening 55 of the pin 34. This surface 53 may be placed on top of a sample film 11 having samples 15 on its opposite surface. By applying a weak vacuum to the hollow pin 34, the sample film 11 may be sucked and stick to the pin 34 such that using the removing device 33 the sample film 11 and the samples 15 sticking thereto may be handled and e.g. transported for subsequent analysis.

(23) In FIG. 8(b) an alternative example of a removing device 33 is shown. In this example, the slanted surface 53′ comprises a concave geometry. By pushing the pin 34 onto a flexible sample film 11 the latter may be slightly deformed and may sealingly abut to the end opening 55. By applying a weak vacuum to the hollow pin 34, the sample film 11 may be sucked and stick to the pin 34 such that using the removing device 33 the sample film 11 and the samples 15 sticking thereto may be handled. In such example, the sample film 11 will be slightly bent due to the concave form of the surface 53′ of the pin 34. Due to such bending, the sample 15 sticking to the sample film 11 may come closer to a middle axis 51 of the pin 34, i.e. a distance d2 between the sample 15 and the middle axis 51 may become smaller than the respective distance d1 in the example shown in FIG. 8(a).

(24) Moreover, the automated system 21 as shown in FIG. 3 further comprises an analyzing device 41 such as an X-ray device and an imaging device 43 such as an X-ray detector for data collection. These devices 41, 43 may be adapted for in-plate-data-collection, particularly when both the sample film 11 and the sealing film 7 of the multi-cell plate 1 are designed to be compatible with X-ray data collection. Furthermore, devices 41, 43 may be adapted for analyzing single harvested samples 15.

(25) The automated system 21 further comprises a freezing unit 45 also denoted as freezing station. The freezing unit provides a cryogenic treatment to single samples 15. Furthermore, it is also possible to treat a multi-cell plate 1 in the freezing unit 45. However, for in-plate-data-collection the multi-cell plates 1 may be preferably transferred to directly to the analyzing device 41 without freezing.

(26) Furthermore, the automated system 21 further comprises a storage unit 47 also denoted as storage station. In the storage unit 47 single harvested samples 15 and multi-cell plates 1 may be stored before analysis.

(27) In FIG. 4 a method for post processing or processing the samples 15 directly in the multi-cell plate 1 is described. In particular, in FIG. 4 a method for supplying a fluid to the samples 15 is described. The fluid may contain chemicals such as possible ligands or cryo protectants. This method may be executed with the automated system shown in FIG. 2 or FIG. 3.

(28) After a crystallisation of the biological material takes place in the cells 5 of the multi-cell plate 1, at least the position of one crystal 15 within one cell 5 is identified/registered in step S08 by the identification device 27 of the automated system 21. The identification may take place automatically e.g. based on markers 17 or on image analysis by a control device 39. Alternatively, a user may identify a cell and the position of the crystals within the cell 5 based on images provided by the identification device 27. This later step can also be performed remotely, for example through a web-based software interface.

(29) In step S12 a depressurization opening could be provided in the sample film 11. In step S13 an opening is provided in the sample film 11 by the cutting device 23 e.g. by photo ablation. In step S18 the fluid unit 29 is connected to the opening. This may e.g. take place by displacing the multi-cell plate 1 to the fluid unit 29. Furthermore, in a step S19 a fluid in a selected amount and composition is supplied at the opening by the fluid unit 29. Then, the solutions may be aspirated through the opening to remove excess liquid. Then, the film area may be cut and the film may be mounted on or attached to the pin by the removing device 33. Then, the crystal may be frozen. Then, the crystal may be stored or directly exposed to x-rays. As shown in FIG. 6 further steps may be included in this method.

(30) This method may be particularly advantageously applied in macromolecular crystallography which often requires treatment of crystals with chemicals. According to known systems and methods this treatment may be a manual and time consuming process requiring skilled operators. The method and the corresponding system 21 of the invention allow full automation of the process of delivering chemicals to crystals. The system 21 and also the method described allow crystals grown on the sample film 11 inside a closed cell 5 to be put in contact with the chemical through a small opening made in the sample film 11. Samples 15 and in particular crystals treated in this way may be directly exposed to x-rays in its original support, i.e. in the multi-cell plate 1. Alternatively, they may be recovered and exposed to x-rays after cryo-cooling for structural analysis.

(31) In FIG. 5 a further method for processing the samples 15 directly in the multi-cell plate 1 is described. In particular, the method in FIG. 5 is a method for extracting a fluid from the samples 15. The methods shown in FIG. 4 and FIG. 5 may be combined as described in FIG. 6.

(32) The first steps S08, S13 and S18 of the method shown in FIG. 5 may be similar to the steps shown in FIG. 4. However, instead of adding a fluid with chemicals to the cell 5 a fluid is extracted e.g. aspirated from the identified cell 5 by the fluid unit 29. The opening provided in step S13 gives access to the solution in which the sample 15 and in particular the crystal is located. During this process the sample 15 remains in its original support, i.e. in the multi-cell plate 1 and the user interaction is minimized because the crystal does not have to be extracted from the multi-cell plate 1. The crystal may then be directly exposed to x-rays in its original support. Alternatively, the crystal may be recovered by excising the sample film area containing the crystal and presented to a cryo stream or to other cryogenic systems.

(33) FIG. 6 shows a flow diagram of a method which combines several automated steps. Some of the steps shown in FIG. 6 may optionally require user interaction. Moreover, several steps of the method in FIG. 6 are executed only optionally. The steps described in connection with FIG. 4 and FIG. 5 are also included in the method shown in FIG. 6.

(34) In a first step S01 biological material 15 is set on a sample film 11 of a multi-cell plate 1. In a second step S02 the cells 5 of the multi-cell plate 1 are sealed by a sealing film 7. In step S03 crystals are grown in the cells 5 e.g. by vapour diffusion crystallization. In step S05 images of the cells 5 of the multi-cell plate 1 are recorded by the identifying device 27 of the system 21. Images can also be taken in an imaging device external to the system and the images may be imported into the system through the software component. Therein, images of all the cells 5 may be acquired. Alternatively, only images of certain areas of the multi-cell plate 1 and thus only images of certain cells 5 may be acquired.

(35) In step S07 the images are taken by the control unit 39 of the system 21 and/or by an operator. The control unit 39 or the operator identifies in step S08 locations of one or multiple crystals in a cell 5. Preferably, steps S07 and S08 can be performed through an external system and recorded positions and images may be provided to the control unit of the system. In case of the identification by the control unit 39 this step may be conducted automatically. e.g. based on markers 17 or on image analysis. After the identification in step S08 an operator or the control unit 27 may select how to process the samples 15 of the multi-plate and which samples 15 are to be processed. These operations may include steps S08b to S08f which are shown in FIG. 7 and which may be executed e.g. after identifying the positions of the crystals in step S08 and before transferring the multi-cell plate 1 in step S09. Alternatively, steps S08b to S08f may be performed after transferring the multi-cell plate 1 in step S09 by a positioning device. Therein, these steps may include specifying a cutting area (S08b) specifying a chemical treatment and which solutions and which volumes are to be used for this treatment (S08c). Furthermore, specifying a position to make an opening (S08d) and specifying whether liquid is to be aspirated (S08e). Moreover, specifying whether crystals are to be analyzed in plates or harvested with the laser and mounted on a pin and whether they are going to be frozen or analyzed without freezing (S08f). At least steps S07, S08 and S08b to S08f may be performed remotely for example through an internet-based software or may be performed earlier and recorded so that all operations are performed at a later time in the system.

(36) As shown in FIG. 6 after the crystal growth the multi-cell plate 1 may be transferred to the system (S09) and operations S08 to S08f executed through the software component of the system. Alternatively, steps S08 to S08f may be introduced (either through the system software or through other software for example a web-based software for remote operation) and recorded. At a later point this information is transferred to the system (S17) and the multi-cell plate is placed in the system (S09) so that the processing may start.

(37) Once the plate is in the system and according to the protocol selected through steps S08 to S08f the sample 11 may be processed in different ways. For example, the plate is placed in the system and the crystal is aligned into the working area (S09), then an opening is made (S13), then solution is aspirated (S23), then the sample may be harvested by a removing device 33 in step S25. For this purpose the film area around the crystal is excised with the laser system (S25a), then the film area containing the crystal is mounted on a pin (S25b) and the harvested crystal is either frozen (S27) in the freezing unit or analyzed directly in the X-ray analysis system (S33). Therein, steps S25a and steps S25b are not shown in the Figures. To perform step S33 precise crystal positions are transferred from the system to the x-ray analysis device in step S31b (not shown in the Figures). If the harvested crystals (S09, S13, S13, S15b, S25a) are frozen (S27) in the freezing unit they may be either analyzed directly with X-rays (S31b, S33), or stored (S29) in the storage unit and analyzed later (S31b, S33).

(38) Multiple variations to this protocol are possible, as indicated in FIG. 7. For example, after making an opening, chemicals may be delivered to crystals (S18) with the solution delivery unit of the fluid unit, then liquids may be aspirated (S23) and the process proceed like shown in FIG. 6 and described above. Solution delivery in step S18 and aspiration in step S23 may be repeated any number of times, for example to deliver multiple solutions of the same or different compositions sequentially before proceeding to steps S25 and further.

(39) In another example, crystals can be harvested without any treatment. This would involve S09, S25a, S25b, S27 (optionally), S29 (optionally) and S33 (optionally, also including (S31b) in this case). In another example, crystals may be directly analyzed in the multi-cell plates. This involves identification of the crytal positions (S08), transfer of the crystal positions relative to the markers 17 to an x-ray analysis device (S31b) and transfer of multi-cell plates to x-ray analysis device and analysis by X-rays (S31). The results of the in-plate-data collection are enhanced by the use of a sample film 11 as described above. It is also possible to remove the liquid without removing the crystal and analyse the crystal inside the plate (S08, S09, S13, S18, S31b, S31). Other possible crystal processing and analysis protocols are indicated in FIG. 7.

(40) In more detail, with reference to FIG. 6 again, after the crystal growth the multi-cell plate 1 may be transferred to a freezing unit 45 and frozen, i.e. cryo processed in step S27. Furthermore, the multi-cell plate 1 may be transferred to a storage unit 47 and stored in step S29. These steps are optional. After cryo treatment and storage or instead of these steps the multi-cell plate 1 may be transferred directly to an analyzing device 41 such as an X-ray device in step S31. In this case, in-plate-data collection takes place by an imaging device 43 in step S33. For example, the sample 15 of a certain cell 5 is analyzed by X-ray diffraction.

(41) Alternatively, a selected sample 15 may be harvested by a removing device 33 in step S25 before analysis. In a further alternative (not shown in the Figures) the sample 15 may be harvested before freezing S27 and before storing S29. For harvesting the multi-cell plate 1 may be brought into position by a positioning plate 37 e.g. above a cutting device 23. Therein, the removing device 33 may be designed as a hollow pin as shown in FIG. 3. While the cutting device 23 cuts the sample film 11 around the selected sample 15 a negative pressure is applied to the sample film 11 below the sample 15. In addition to the negative pressure an adhesive e.g. a glue may be applied at the tip of the removing device 33 for ensuring a secure attachment of the sample 15 to the removing device 33. After harvesting the sample 15 may be stored, frozen and/or analyzed.

(42) After crystal growth and before the steps S25 to S33 the samples 15 may be processed in their original support, i.e. in the multi-cell plate 1. In step S09 the multi-cell plate 1 may be transferred by the positioning device 25 to different units and devices of the system 21. For example, the positioning device 25 may bring the multi-cell plate 1 into correct position and orientation above the cutting device 23 or transfer the multi-cell plate 1 to the fluid unit 29. Furthermore, steps similar to the steps described in relation to FIG. 4 and FIG. 5 may follow step S09.

(43) In step S11 a position, shape and size of an opening to be provided by the cutting device 23 at an identified cell 5 is selected. The selection may take place automatically be the control device 39 or by an operator. In step S13 the opening in the sample film 11 is provided by the cutting device 23 e.g. below the sample 25. Through the opening chemicals may be administered to the sample 15. Furthermore, a fluid such as the sample solution may be extracted through the opening. In step S15 the composition of the fluid to be supplied to the a certain cell 5 is selected. In step S17 the selected information is transferred to the fluid unit 29. Furthermore, in step S18 the fluid unit 29 is connected to the opening and a fluid in a selected amount and composition is supplied at the opening by the fluid unit 29 in step S19. This process, i.e. steps S09 to S19 may be repeated in step S21 with different chemicals such as different potential ligands and finally with a cryo protectant.

(44) Alternatively, after providing the opening in the sample film 11 in step S13 a fluid may be extracted from the cell 5 by applying suction via the fluid unit 29 in step S23 as explained in FIG. 5. As shown in FIG. 6 step S23 may optionally be executed after chemicals are supplied to the sample 15 in step S19. Subsequently to supplying fluids to the cell 5 and/or extracting fluids from the cell 5 steps S25 to S33 may be executed.

(45) It has to be noted that embodiments of the invention are described with reference to different subject matters. In particular, some embodiments are described with reference to method type claims whereas other embodiments are described with reference to the device or system type claims. However, a person skilled in the art will gather from the above and the following description that, unless otherwise notified, in addition to any combination of features belonging to one type of subject matter also any combination between features relating to different subject matters is considered to be disclosed with this application. However, all features can be combined providing synergetic effects that are more than the simple summation of the features.

(46) While the invention has been illustrated and described in detail in the drawings and foregoing description, such illustration and description are to be considered illustrative or exemplary and not restrictive. The invention is not limited to the disclosed embodiments. Other variations to the disclosed embodiments can be understood and effected by those skilled in the art in practicing a claimed invention, from a study of the drawings, the disclosure, and the dependent claims.

(47) Furthermore, the term “comprising” does not exclude other elements or steps, and the indefinite article “a” or “an” does not exclude a plurality. The mere fact that certain measures are re-cited in mutually different dependent claims does not indicate that a combination of these measures cannot be used to advantage. Any reference signs in the claims should not be construed as limiting the scope.

LIST OF REFERENCE SIGNS

(48) 1 multi-cell plate 3 body 5 cell 7 sealing film 9 first side of body 11 sample film 13 second side of body 15 sample/biological material for crystallization 17 marker 19 precipitate reservoir 21 automated system 23 cutting device 25 positioning device 27 identification device 29 fluid unit 31 fluid with chemicals 33 removing device/mounting unit 34 pin 35 robotic arm 37 plate support 39 control device 41 analyzing device 43 imaging device 45 freezing unit 47 storage unit 49 axis through sample 51 middle axis of pin S01 Incubating biological material on a sample film of a multi-cell plate S02 Sealing the cells of the multi-cell plate by a sealing film S03 Growing crystals S05 Recording images of the cells of the multi-cell plate by the identification device S07 Providing the images to a control unit and/or to an operator S08 Identifying positions of crystals in a cell e.g. based on markers or on image analysis by the identification device S08b Specifying an area to be cut by the cutting device, particularly by a laser S08c Specifying whether chemicals are to be delivered (with the solution delivery unit), which ones, and in which quantities S08d Specifying a position of the opening S08e Specifying whether liquid is to be aspirated S08f Specifying whether crystals are to be analyzed in plates or harvested with the laser and mounted on a pin and whether they are going to be frozen or analyzed without freezing S09 Transferring the multi-cell plate by a positioning device S11 Selecting a position, shape and size of the opening to be provided by the cutting device at a first cell (if not already done in S08d) S12 providing a depressurization opening in the sample film S13 Providing the opening in the sample film by the cutting device S15 Selecting the composition of the fluid to be supplied to the first cell (if not already done in S08c) S17 Transmitting the selected information to the system S18 Connecting the fluid unit to the opening S19 Supplying a fluid in a selected amount and composition at the opening by the fluid unit S21 Repeating steps S09 to S19 S23 Extracting fluid from the cell by applying suction via the aspiration unit of the fluid unit S25 Harvesting the sample by a removing device S25a Cutting a sample film area around a crystal or around a group of crystals S25b Mounting the film area onto a pin with the removing device S27 Transferring the multi-cell plate or the mounted crystal to a freezing unit and freezing S29 Transferring the multi-cell plate or the mounted crystal to a storing unit and storing S31 Transferring the multi-cell plate to an analysis device S31b Transferring crystal positions to an x-ray analysis device S33 Data collection/Analyzing the sample by an imaging device