METHOD FOR PRODUCING AT LEAST ONE CLOSED REGION ON A CARRIER SURFACE OF A CARRIER

Abstract

The invention relates to a method for producing at least one closed region on a carrier surface of a carrier, wherein the method comprises the following steps: a. adding a first fluid comprising at least one cell and/or at least one particle to the carrier surface and b. adding a second fluid, wherein the second fluid is immiscible with the first fluid and at least partially covers the first fluid and c. acquiring at least one item of cell information and/or of at least one item of particle information and d. producing the closed region on the basis of the at least one acquired item of cell information and/or the at least one acquired item of particle information.

Claims

1. A method for producing at least one closed region (2) on a carrier surface (7) of a carrier (3), wherein the method comprises the following steps: a. adding a first fluid (4) comprising at least one cell (1) and/or at least one particle to the carrier surface (7) and b. adding a second fluid (5), wherein the second fluid (5) is immiscible with the first fluid (4) and at least partially covers the first fluid (4) and c. acquiring at least one item of cell information and/or of at least one item of particle information and d. producing the closed region (2) on the basis of the at least one acquired item of cell information and/or the at least one acquired item of particle information.

2. The method according to claim 1, wherein a. the region (2) is produced in that the first fluid (4) is displaced in such a way that part of the second fluid (15) wets the carrier surface (7) or b. the region (2) is produced in that the first fluid (4) is displaced in such a way that part of the second fluid (15) wets the carrier surface (7) and the part of the second fluid (15) forms a closed line.

3. The method according to claim 1, wherein a. a plurality of regions (2) are produced, wherein the regions (2) have the same or different cross-sections and/or b. a plurality of regions (2) are produced which differ from one another in their formation along a normal to the carrier surface (7).

4. The method according to claim 1, wherein a. on the basis of the item of cell information and/or the item of particle information, it is determined where the at least one region (2) is produced and/or b. the region (2) has at least one cell (1) and/or at least one particle.

5. The method according to claim 1, wherein before the region (2) is produced a. on the basis of the acquired item of cell information and/or item of particle information, the carrier (3) is moved or b. on the basis of the acquired item of cell information and/or item of particle information, the carrier (3) is moved in such a way that the at least one cell (1) and/or the at least one particle is moved into a new position or c. on the basis of the acquired item of cell information, the carrier is shaken or vibrated and/or the first fluid is mixed so that the at least one cell (1) and/or the at least one particle is moved into a new position.

6. The method according to claim 1, wherein a. at least one remaining region (8) having no cell (1) is removed and/or b. a reagent is introduced into the region (2) having in particular a cell or a particle and/or c. the region (2) having in particular a cell or a particle is at least partially sucked out or filled up.

7. The method according to claim 1, wherein the region (2) is divided into at least two sub-regions (9) by displacing the first fluid (4) of the region (2) in such a way that part of the second fluid (5) of the region (2) wets the carrier surface (7).

8. The method according to claim 1, wherein the first fluid (4) and the second fluid (5) are displaced via a displacement agent (6).

9. The method according to claim 8, wherein the displacement agent (6) and/or the carrier (3) are moved in order to displace the first fluid (4) and/or the second fluid (5).

10. The method according to claim 8, wherein a. the displacement agent (6) has a solid body, wherein the first fluid (4) is displaced by the solid body or b. the first fluid (4) is displaced by applying a gas from the displacement agent (6) to the first fluid (4).

11. The method according to claim 8, wherein a. to produce the region (2) the displacement agent (6) is moved exclusively in the direction of the carrier surface (7) or the carrier (3) is moved exclusively in the direction of the displacement agent (6) or b. to produce the region (2) the displacement agent (6) and/or the carrier (3) is moved in at least one direction parallel to the carrier surface (7).

12. A device (10) for carrying out the method according to claim 1.

13. A device (10) for producing at least one closed region (2) on a carrier surface (7) of a carrier (3) for receiving a first fluid (4) and a second fluid (5), wherein the second fluid (5) is immiscible with the first fluid (4) and at least partially covers the first fluid (4), having an optical acquisition device (25) for acquiring at least one item of cell information and/or of at least one item of particle information and a displacement agent (6), by means of which, on the basis of the at least one acquired item of cell information and/or the at least one acquired item of particle information, the closed region (2) can be produced.

14. The device according to claim 13, wherein the displacement agent (6) is designed and intended to displace the first fluid (4) in order to produce the region (2), so that part of the second fluid (15) wets the carrier surface (7).

15. The device (10) according to claim 13, wherein a. the acquisition device (25) has an imaging device (11) for producing an image of the carrier surface (7) or b. the acquisition device (25) has an imaging device (11) for producing an image of the carrier surface (7) and an evaluation device (12), which determines the item of cell information and/or item of particle information on the basis of the image.

16. The device (10) according to claim 15, wherein the device (10) has a receptacle (13) for receiving the carrier (3), wherein the imaging device (11) and the displacement agent (6) are mutually opposite one another with respect to the receptacle (13).

17. The device (10) according to claim 13, wherein a. the displacement agent (6) has a hydrophobic solid body and/or b. the displacement agent (6) has a pattern element (27) at its end which can be brought into contact with the first fluid (4) and/or second fluid (15) and/or c. the displacement agent (6) has a gas outlet opening.

18. The device (10) according to claim 13, wherein a. a removal device (14) for removing the remaining region (8) of the first fluid (4) having no cells and/or for removing the region (2) or b. a removal device (14) for at least partially aspirating the remaining region (8) of the first fluid (4) having no cells and/or for at least partially sucking out the region (2).

19. A system having a device (10) according to claim 13 and a carrier (3), which receives the first fluid (4) and the second fluid (5), wherein the second fluid (5) is immiscible with the first fluid (4) and at least partially covers the first fluid (4).

Description

BRIEF DESCRIPTION OF THE DRAWING VIEWS

[0048] The subject of the disclosure is shown schematically in the figures, wherein elements that are the same or have the same effect are mostly provided with the same reference symbols. In the drawings:

[0049] FIG. 1 shows the carrier with a first fluid and a second fluid,

[0050] FIG. 2 shows a side view of part of the device according to the disclosure before a region is produced,

[0051] FIG. 3 shows a side view of part of the device according to the disclosure, in which a region having a cell is produced according to a first operating mode of the displacement agent,

[0052] FIG. 4 shows a side view of part of the device according to the disclosure, in which a region having a cell is produced according to a second operating mode of the displacement agent,

[0053] FIG. 5 shows a side view of part of the device according to the disclosure after a plurality of regions have been produced, each having a cell,

[0054] FIG. 6 shows a plan view of a carrier after a plurality of regions have been produced and

[0055] FIG. 7 shows a side view of part of the device according to the disclosure with a region having a cell.

DETAILED DESCRIPTION

[0056] FIG. 1 shows a carrier 3 which is designed as a container. The carrier 3 contains a first fluid 4 and a second fluid 5 which is immiscible with the first fluid 4. The first fluid 4 contains a plurality of cells 1. Designs are also conceivable in which the first fluid 4 alternatively or additionally contains particles. The first fluid 4 and the second fluid 5 are added to the carrier 3. The first fluid 4 can preferably be added before the second fluid 5. The second fluid 5 is arranged on the first fluid 4 and completely covers this within the carrier 3.

[0057] FIG. 2 shows a side view of part of the device 10 according to the disclosure before a region is produced. As can be seen from FIG. 2, the carrier 3 is arranged on a receptacle 13 of the device 10. The device 10 has a displacement agent 6 which can be moved along the directions x, z, y. In addition, the device 10 has an optical acquisition device 25. The acquisition device has an imaging device 11, by means of which an optical image of the carrier 3, in particular a carrier surface 7, can be produced.

[0058] In addition, the acquisition device 25 has an evaluation device 12. The imaging device 11 is electrically connected to the evaluation device 12. The evaluation device 12 serves to determine items of cell information, such as the position of the cells 1 located in the first fluid 4 on the carrier surface 7. The evaluation device 12 can be electrically connected to a control device, not shown in detail. The imaging device 11 and the displacement agent 6 can lie opposite one another with respect to the receptacle 13 of the device 10 or can be arranged offset from one another.

[0059] The device 10 can have a moving device 26 which is controlled by the control device. The moving device 26 can move the acquisition device 25 and/or the displacement agent 6 and/or the carrier 3 along the directions x, y, z. In particular, the moving device 26 can move the displacement agent 6 and/or the carrier 3 on the basis of the item of cell information determined by the evaluation device 12 in order to produce a region described in more detail below.

[0060] The displacement agent 6 is designed as a hydrophobic pin. The displacement agent 6 has a rounded end at its end facing towards the second fluid 5.

[0061] FIG. 3 shows a side view of part of the device 10 according to the disclosure, in which a closed region 2 having a cell is produced according to a first operating mode of the displacement agent 6. Before the region 2 is produced, the item of cell information is determined by means of the acquisition device 25. In particular, the regions of the carrier surface 7 in which the cells 1 are arranged are determined.

[0062] The displacement agent 6 is then moved in such a way that it creates the closed region 2 which includes the cell 1. To produce the region 2, the displacement agent 6 is first moved along the direction y to the carrier surface 7. After wetting the carrier surface 7 by a part of the second fluid 5, the displacement agent 6 is moved along the directions z and x in order to produce the closed region 2. When the displacement agent 6 moves along the directions z and x, the first fluid 4 is also displaced by the displacement agent 6, so that the second fluid 5 can wet the carrier surface 7.

[0063] In this operating mode, the displacement agent 6 presses the part of the second fluid 5 in the direction of the carrier surface 7 until the part of the second fluid 5 wets the carrier surface 7. In this operating mode, the displacement agent 7 does not come into direct contact with the carrier surface 7. As can be seen from FIG. 4, the part of the second fluid 5 is arranged between the displacement agent 6 and the carrier surface 7.

[0064] The region 2 is fluidically separated from a remaining section 18 by means of the part of the second fluid 15, wherein the remaining section 18 has the first fluid 4 with the plurality of cells 1. The fluidic separation takes place such that the first fluid 4 located in the region 2 is not fluidically connected to the first fluid 4 located in the remaining section 18.

[0065] FIG. 4 shows a side view of part of the device 10 according to the disclosure, in which a closed region 2 having a cell 1 is produced according to a second operating mode of the displacement agent 6. In this operating mode, the displacement agent 6 is moved in such a way that, when it moves along the direction y, it passes through the second fluid 5 and displaces the first and second fluids 4, 5. The displacement agent 6 comes into direct contact with the carrier surface 7.

[0066] After the displacement agent 6 contacts the carrier surface 7, it is moved in the direction x and/or z in order to produce the closed region 2. When the displacement agent 6 is moved along the x and/or z direction, the first fluid 4 is also displaced. After the displacement agent 6 has moved, part of the second fluid 5 flows into the portion of the first fluid 4 displaced by the displacement agent 6 and thus wets the carrier surface 7.

[0067] FIG. 5 shows a side view of part of the device according to the disclosure after a plurality of regions 2 have been produced, each having a cell 1. In particular, FIG. 5 shows that precisely three regions 2, namely a first region 22, a second region 16 and a third region 17, were produced by means of the device 10. In addition, the carrier 3 contains a remaining region 8 that does not contain any cells.

[0068] The individual regions 2 are fluidically separated from one another and from the remaining region 8 by part of the second fluid 15. In particular, the first region 22 is fluidically separated from the remaining region 8 by a first part of the second fluid 23. The remaining region 8 is also fluidically separated from the second region 16 by means of a second part of the second fluid 19. The second region 16 is further fluidically separated from the third region 17 by means of a third part of the second fluid 20.

[0069] To produce the regions 2, as already described in FIG. 3, after the two fluids 4, 5 have been introduced into the carrier 3 by means of the imaging device 11, an image of the carrier 3, in particular the carrier surface 7, or, by means of image stacking, of the entire fluid volume is produced. The evaluation device 12 determines the item of cell information, in particular the position of the cells 1 located in the first fluid 4, on the basis of the produced image.

[0070] To produce the first region 22, the displacement agent 6 is moved on the basis of the determined positions of the cells 1 into a position relative to the carrier 3 and, as shown in FIG. 3 or 4, is lowered along the direction y in the direction of the carrier surface 7.

[0071] Here, the rounded end of the displacement agent 6 displaces the first fluid 4 in such a way that the first part of the second fluid 23 wets the carrier surface 7. The displacement agent 6 is then moved along the directions z, x, which run parallel to the carrier surface 7, to produce the first region 22.

[0072] The first region 22 is fluidically separated from the rest of the first fluid 4 located within the carrier 3 by means of the first part of the second fluid 23. The first region 22 is thus delimited by the carrier surface 7, a side wall of the carrier 3, the first part of the second fluid 23 wetting the carrier surface 7 and the second fluid 5 wetting the first region 22. After the first region 22 has been produced, the displacement agent 6 is moved away from the carrier surface 7 in the direction y. The displacement agent 6 is then moved into a different position relative to the carrier 3, starting from which the second region 16 shown in FIG. 5 is produced. After completion of the second region 16, the third region 17 is produced.

[0073] The second and third regions 16, 17 are produced analogously to the first region 22, in that the displacement agent 6 displaces the first fluid 4 in such a way that the corresponding part of the second fluid 19, 20 wets the carrier surface 7. In addition, the displacement agent 6 is moved parallel to the carrier surface 7. Even when the displacement agent 6 moves parallel to the carrier surface 7, the first fluid 4 is displaced in such a way that the corresponding part of the second fluid 19, 20 wets the carrier surface 7.

[0074] The device 10 also has a removal device 14. The remaining region 8 can be sucked out by means of the removal device 14. In this case, the removal device 14 serves to remove the remaining region 8 which does not have any cells. As an alternative or in addition, the removal device 14 can serve to suck out the regions 2. The suction takes place after a predetermined period of time or inspection, so that cell growth has taken place in the respective regions 2.

[0075] FIG. 6 shows a plan view of a carrier 3 after a plurality of regions 2, namely the first region 22, the second region 16 and the third region 17, have been produced. The arrangement of the cells 1 in the carrier 3 shown in FIG. 6 differs from the arrangement of the cells 1 in the carrier 3 shown in FIGS. 1 to 5.

[0076] As can be seen from FIG. 6, the three regions 2 differ in their cross-section. In addition, the three regions 2 differ from one another in their design and volume. The first region 22 has a circular shape, the second region 16 has a rectangular shape and the third region 17 has a triangular shape. The remaining region 8 was removed by means of the removal device 14.

[0077] The second region 16 can be divided into two sub-regions 9. This can occur by the displacement agent 6 displacing the first fluid 4 of the second region 16. As a result, a fourth part of the second fluid 21 wets the carrier surface 7. The fourth part of the second fluid 21 separating the two sub-regions 9 is shown in FIG. 6 by a dashed line.

[0078] The regions 2 shown in FIG. 6 can have been produced by means of a method shown in FIGS. 2-5. This means that the displacement agent 6, after it has already pressed the respective part of the second fluid 15 against the carrier surface 7 or is itself in direct contact with the carrier surface 7, is moved along the x, y direction. Alternatively, the regions shown in FIG. 6 can be produced according to the method described below in connection with FIG. 7.

[0079] FIG. 7 shows a side view of part of the device 10 according to the disclosure with a region 2 having a cell 1. The device 10 differs from the device 10 shown in FIG. 3 or FIG. 4 in the design of the displacement agent 6. Thus, the displacement agent 6 shown in FIG. 7 has a pattern element 27 at its end facing the carrier surface 7, by means of which, for example, a cross-section circular region 2 can be produced. The pattern element 27 is designed as a hollow cylinder.

[0080] To produce the closed region 2, the moving device 26 moves the displacement agent 6 exclusively in direction y. The displacement agent 6 thus presses the part of the second fluid 15 in the direction of the carrier surface 7 until the same wets the carrier surface 7. Alternatively, the displacement agent 6 can be operated in a similar way to the embodiment shown in FIG. 4, such that the displacement agent 6 displaces the first fluid 4 such that it comes into direct contact with the carrier surface 7. After the displacement agent 6 contacts the carrier surface 7, this is moved in the opposite direction in direction y. A part of the second fluid 5 wets the carrier surface 7, so that a fluidic separation between the region 2 and the remaining section 18 of the first fluid is realised.

[0081] By using the displacement agent 6 shown in FIG. 6, it is not necessary to move the displacement agent 6 in the directions x, z in order to produce the closed region 2. The region can namely be produced solely by moving the displacement agent 6 exclusively in direction y.

LIST OF REFERENCE NUMBERS

[0082] 1 Cell

[0083] 2 Region

[0084] 3 Carrier

[0085] 4 First fluid

[0086] 5 Second fluid

[0087] 6 Displacement agent

[0088] 7 Carrier surface

[0089] 8 Remaining region having no cells

[0090] 9 Sub-region

[0091] 10 Device

[0092] 11 Imaging device

[0093] 12 Evaluation device

[0094] 13 Receptacle

[0095] 14 Removal device

[0096] 15 Part of the second fluid

[0097] 16 Second region

[0098] 17 Third region

[0099] 18 Remaining section

[0100] 19 Second part of the second fluid

[0101] 20 Third part of the second fluid

[0102] 21 Fourth part of the second fluid

[0103] 22 First region

[0104] 23 First part of the second fluid

[0105] 25 Acquisition device

[0106] 26 Moving device

[0107] 27 Pattern element

[0108] x, y, z Direction