JP2006094783A - Cell supply, exhaust and trap apparatus, and method for supplying, exhausting and trapping cell - Google Patents

Abstract

<P>PROBLEM TO BE SOLVED: To provide cell supply, exhaust and trap apparatus and a method for supplying, exhausting and trapping cells, so as to retain much cells in a short time and to efficiently recover the retained cells after retaining. <P>SOLUTION: A cell supply mechanism 1 for supplying a cell suspension 2, a liquid transferring mechanism 3 for supplying a liquid 4 containing no cell, a trapping chip 5 having a flow path for flowing the cell suspension 2 and the liquid 4 and a through hole 7 trapping cells in the cell suspension 2, a suction mechanism 8 connected with the trapping tip 5 and for controlling suction amount through the through hole 7, a useless cell recovering mechanism 9 for recovering useless cells not trapped at the through hole 7, and a trapped cell recovering mechanism 10 for recovering trapped cells are at least equipped. <P>COPYRIGHT: (C)2006,JPO&NCIPI

Description

  The present invention relates to a cell supply / discharge device and a cell supply / discharge method, and mainly relates to the introduction of substances into cells or the course of cells in the field of life science, particularly in fields related to regenerative medicine and genomic drug discovery. When observing, a plurality of cells are transported / captured on the flow path, and a cell supply / discharge device and a cell supply / discharge / capture characteristic of the configuration are used to collect uncaptured cells and captured cells separately. It is about the method.

  In recent years, in fields such as regenerative medicine and genomic drug discovery, opportunities to use cells into which genes and drugs have been introduced have increased, and many substance introduction technologies have been used for research purposes. There are different requirements for research purposes.

In particular, the main requirements specific to medical use are that the type of cell and the type of introduced substance are not limited, the introduction efficiency is high, and that a large amount of substance-introduced cells can be supplied. Mass supply of substance-introduced cells is particularly important, and it is said that 10 ⁵ to 10 ⁶ cells are required at a time in regenerative medicine.

  In addition, there are various methods for introducing a substance. Among them, the microinjection method (see, for example, Patent Documents 1 and 2) has a high success rate of substance introduction, and does not select a combination of cells and introduced substances. Therefore, it is considered the most reliable method for introducing substances.

  When producing a substance-introduced cell using such a microinjection method, it is necessary to hold the cell in a predetermined location. For this purpose, various methods have been proposed. For example, an inverted quadrangular pyramid is formed on a silicon substrate. It has been proposed that the holes are arranged in a two-dimensional matrix, and one cell is isolated and held in each inverted quadrangular hole (see, for example, Patent Document 3).

Moreover, in said patent document 3, providing the slit for draining in the bottom part of an inverted square pyramid hole is also proposed.
JP 05-192171 A Japanese Patent Laid-Open No. 06-343478 JP-A 63-129980

  However, when microinjection, cell electrical organization experiments, etc. are performed using the cell holding device shown in Patent Document 3, cells can be held, but one cell is surely inserted into each inverted quadrangular hole. There is a problem that a specific method for efficiently accommodating has not been established.

  In addition, since individual cells are isolated, there is a problem that it is difficult to collect cells when it is necessary to collect and culture after the experiment. There is a problem that it cannot be applied directly to industrial applications such as medicine.

  On the other hand, there are also cell-adhesion fixation methods using poly-L-lysine, lectins, etc., but there are concerns about toxicity to cells, and in addition, it is difficult to detach and recover cells after fixation. There is.

  Therefore, an object of the present invention is to hold a large amount of cells in a short time and to efficiently recover the cells held after the holding.

FIG. 1 is a diagram illustrating the basic configuration of the present invention. Means for solving the problems in the present invention will be described with reference to FIG.
In order to solve the above problems, the present invention provides a cell supply mechanism 1 for supplying a cell suspension 2, a liquid supply mechanism 3 for supplying a liquid 4 not containing cells, A capture chip 5 having a flow path 6 for flowing the cell suspension 2 and the liquid 4 and a through-hole 7 for capturing cells in the cell suspension 2, and a suction amount from the through-hole 7 connected to the capture chip 5 It comprises at least a suction mechanism 8 for controlling, an unnecessary cell recovery mechanism 9 for recovering unnecessary cells that have not been captured by the through-hole 7, and a captured cell recovery mechanism 10 for recovering captured cells.

By adopting the above configuration, it becomes possible to separately collect captured cells and uncaptured cells, and improve the efficiency of analysis work after collection.
In addition, since the cells are captured by aspiration, the capturing means itself does not adversely affect the cells, and the overall configuration of the device is compact so that even when the number of samples is small, reliable capture is possible. It can be carried out.

In this case, the trapping chip 5 has an inflow portion for flowing the cell suspension 2 and the liquid 4 through the flow channel 6 at one end of the flow channel 6, and a cell suspension from the flow channel 6 at the other end of the flow channel 6. It is desirable to provide a plurality of through-holes 7 with a diameter that the cells in the cell suspension 2 cannot pass along the flow path 6 while providing the collection unit for collecting the turbid liquid 2 and the liquid 4. A large number of cells can be captured on the capture chip 5 in a short time.
In addition, the cell can be captured without passing through the through-hole 7 by setting the diameter of the through-hole 7 so that the cell cannot pass therethrough.

  In addition, the flow path 6 provided in the capture chip 5 may be a single meandering flow path 6, or a plurality of parallel flow paths 6 and a common inflow portion at one end of each flow path 6. A branch type flow path 6 having a connection flow path connected to a common recovery section at the other end of each flow path 6, and having a single flow path 6. On the other hand, the cells can be collected in order, and on the other hand, when the branched channel 6 is used, the cells can be captured in a short time by processing the cells in parallel.

  In addition, it is desirable to make the inner wall of the flow channel 6 provided in the capture chip 5 more hydrophilic than the base material of the capture chip 5 and make the surface other than the inner wall of the flow channel 6 of the capture chip 5 more hydrophobic than the base material. Thereby, the cell suspension 2 can be flowed only in the channel 6 without sealing the channel 6.

  In this case, the inner wall of the flow channel 6 may be coated with a hydrophilic resin from the base material of the capture chip 5, and the surface other than the inner wall of the flow channel 6 of the capture chip 5 may be coated with a hydrophobic resin from the base material. .

  Moreover, it is desirable that the liquid 4 containing no cells is a liquid 4 having a heavier specific gravity than the cells, so that the cells settled on the bottom of the flow path 6 can be floated and can be easily transported.

  The liquid 4 containing no cells is preferably the liquid 4 having the same components as the medium constituting the cell suspension 2 and having a higher salt concentration than the medium, and the basic components are the same. So it will not adversely affect the captured cells.

  In addition, by providing such a cell supply / discharge device with an injection mechanism for injecting a substance into the captured cell, it is possible to efficiently perform recovery when culturing the substance-introduced cell.

  In addition, when the above-described supply / discharge / capture of cells is performed, the cell suspension 2 is allowed to flow from one end of the flow path 6 having a plurality of through holes 7, and the cells in the cell suspension 2 are passed through the through holes 7. The step of capturing by suction, the step of flowing the liquid 4 having a higher specific gravity than the cells from one end of the flow channel 6 to float the cells that could not be captured and collecting the cells at the other end of the flow channel 6, and injecting the substance into the captured cells The step of performing either injection or follow-up, suction is canceled after the injection or follow-up, and the liquid 4 having a higher specific gravity than the cells is flowed from one end of the flow path 6 to float the cells and collected at the other end of the flow path 6 The steps may be performed sequentially.

  As described above, according to the present invention, cells can be captured at high throughput, and captured cells and uncaptured cells can be collected separately and with high efficiency, thereby enabling regenerative medicine. And microinjection devices in industrial applications such as genome drug discovery.

The present invention begins with
(1) The cell suspension is supplied from the cell supply mechanism to the inflow portion of the capture chip.
The supply method to the inflow part is that the pipette-like tubule is connected to the cell supply mechanism, and the cell supply mechanism is connected to the capture chip via the tube even if the cell suspension is dropped to the inflow part. A suspension may be supplied.
At this time, by making the flow path of the capture chip hydrophilic and making the portion other than the flow path hydrophobic, the cell suspension can be flowed only in the flow path without sealing the flow path.

Then
(2) A cell is trapped in the through hole in the flow path using a suction mechanism.
The outlet of the through-hole is connected to a suction mechanism, and captures cells by suction pressure from the through-hole. In this case, by setting the diameter of the through-hole to a size that prevents passage of cells, The cell can be captured without passing through the through hole.

Then
(3) Using the liquid feeding mechanism, the cells that have not been captured are removed from the capture chip.
At this time, a liquid having a specific gravity different from that of the cells is supplied from the liquid feeding mechanism to the inflow portion, and the uncaptured cells are floated and conveyed to the outflow portion of the capture chip.
(4) Using an unnecessary cell recovery mechanism, recover the uncaptured cells that have flowed to the outflow part.

Then
(5) Perform operations such as injection and follow-up on the captured cells. After the operation is completed, the suction pressure is released using the suction mechanism to release the captured cells.

Then
(6) Using the liquid feeding mechanism again, the released cells are conveyed to the outflow part of the capture chip. At this time, a liquid having a specific gravity different from that of the cells is supplied from the liquid feeding mechanism to the inflow portion, and the captured cells are floated and conveyed to the outflow portion of the capture chip.
(7) Collect the captured cells that have been transported using the capture cell recovery mechanism.

Here, with reference to FIG.2 and FIG.7, the cell supply-discharge / capture apparatus of Example 1 of this invention is demonstrated.
See Figure 2
FIG. 2 is a conceptual configuration diagram of the cell supply / discharge / capture device according to the first embodiment of the present invention. The cell suspension 13 includes a pipette-like capillary tube 12 and cells 15 are suspended from the capillary tube 12 in a medium 14. Cell supply device 11 for supplying the capture chip 30 to the capture chip 30, and the liquid supply device 16 for supplying the carrier liquid 18 having a heavier specific gravity than the culture medium from the narrow tube 17 to the capture chip 30, and the supplied cell suspension. 13, a trapping chip 30 that sucks and captures the cells 15 by the suction device 23, a container 19 that houses the capture chip and has an opening 20 for suction, holds the container 19, and is connected to the suction device 23. A chip holder 21 having a suction device 23, a suction device 23 for capturing the cells 15, a cell observation device 40 for observing the captured cells 15; The basic configuration of the capture cell collection device 26 unwanted cell harvester 24, the treated cells 15 working injection or the like for collecting and recovering through the capillary 27 through is formed.

  In this case, the cell observation device 40 includes an image processing device 43 including a light source 41 and an objective lens 42. The cell observation device 40 constantly observes the capture chip 30 to detect the capture state of the cells 15 and the cells. Follow up the division.

3 and FIG. 4 are schematic plan views of the capture chip 30, and FIG. 4 is a schematic cross-sectional view of the capture chip 30. The silicon substrate 31 is formed using a normal photolithography process. An inflow portion 32 is formed at one end, an outflow portion 34 is formed at the other end, and a meandering flow path 33 connecting the inflow portion 32 and the outflow portion 34 is formed.

  In addition, a suction through hole 35 is provided at the bottom of the flow path 33, and an opening 36 connected to the through hole 35 is formed on the back surface of the silicon substrate 31, and cells are sucked from the through hole 35 through the opening 36. To do.

In addition, the side wall and the bottom surface of the flow path 33 are coated with a hydrophilic resin 37, for example, HydroCell (trade name, manufactured by Cellseed) to perform hydrophilic treatment, and the surface of the silicon substrate 31 other than the flow path 33 is hydrophobic. The resin 38, for example, an acrylic resin is coated and subjected to hydrophobic treatment.
At this time, care must be taken so that the through hole 35 is not blocked by the resin.

Although the width of the flow path 33 depends on the size of the cells to be captured, when the cell size is about 10 to 15 μm, for example, the width is 50 μm and the depth is 50 μm, and the length of the straight portion is about 500 μm. To do.
Further, the through holes 35 provided at the bottom of the flow path 33 have a diameter of 2 μm, for example, and the through holes 35 are arranged at an interval of 50 μm, for example.

See Figure 5
FIG. 5 is an explanatory diagram of the arrangement configuration of the capture chip or the chip holder. The capture chip 30 is arranged and aligned with, for example, the opening 19 provided in the container 19 with respect to the container 19. Fix it.
In addition, the container 19 in this case is for making the handling easy while preventing the liquid from scattering to the surroundings, and is made of polystyrene having excellent chemical resistance like the petri dish used for culture.

  In addition, the container 19 is arranged and fixed with respect to the tip holder 21 so that the opening 20 is aligned with the connection part 22 with the suction device 23. The provided connection member 28 is connected.

See FIG. 6 and FIG.
FIG. 6 is an operation flow diagram of the cell supply / discharge / capture device of Example 1 of the present invention, and FIG. 7 is an explanatory diagram of the cell capture / transport, release / transport state,
A. The cell suspension 13 is supplied from the cell supply device 11 to the inflow portion 32 provided in the capture chip 30.
In this case, the cell supply device 11 controls the amount and the fluid pressure of the cell suspension 13 to drop from the pipette-like capillary 12.

Then
B. Using the suction device 23, the cells 15 are captured in the plurality of through holes 35 provided in the flow path 33.
In this case, the diameter of the through-hole 35 is 2 μm, for example, and is not discharged from the through-hole 35 that is sufficiently small with respect to the cell diameter of 10 to 15 μm.

Then
C. Using the liquid feeding device 16, the carrier liquid 18 is supplied from the thin tube 17 to the inflow part 32 of the capture chip 30, and the uncaptured cells 15 are floated and conveyed to the outflow part 34.
Here, as the carrier liquid 18, a liquid in which the salt concentration of the balanced salt solution (BBS) used as the medium 14 is increased to make it heavier than the specific gravity of the cells 15 is used.

Then
D. The cells 15 conveyed to the outflow part 34 are recovered from the thin tube 25 using the unnecessary cell recovery device 24.

Then
E. After the follow-up of the trapped cells 15 such as the state of cell division, the suction pressure by the suction device 23 is released, and the trapped cells 15 are released.

Then
F. Again, using the liquid feeding device 16, the carrier liquid 18 is supplied from the thin tube 17 to the inflow portion 32 of the capture chip 30, and the released cells 15 are conveyed to the outflow portion 34.

Then
G. By recovering the observed cells 15 transported to the outflow part 34 from the capillary tube 27 using the capture cell recovery device 26, a series of flows is completed.
Thereafter, the collected observed cells 15 are dispensed into, for example, a 96-well plate in which 96 concave portions are provided in a two-dimensional matrix on a polycarbonate substrate and stored and cultured in small amounts.

  As described above, in the first embodiment of the present invention, since the capture chip 30 provided with the plurality of capture through holes 35 along the flow path 33 is used, unnecessary cells and captured cells are collected separately. It can be performed in a short time, and can be applied not only to a laboratory but also to a microinjection apparatus or the like in industrial applications such as regenerative medicine and genome drug discovery.

  In addition, by making the flow path 33 of the capture chip 30 hydrophilic and making the portion other than the flow path 33 hydrophobic, only the flow path 33 is suspended only in the flow path 33 without sealing the flow path 33. The turbid liquid 13 can be flowed, and the usability is improved.

  Moreover, since the flow path 33 provided in the capture chip 30 is constituted by one meandering flow path, the captured cells can be collected one by one in order.

Next, with reference to FIG. 8, the cell supply / discharge / capture device of Example 2 of the present invention will be described. However, only the configuration of the flow path provided in the capture chip is different, and other configurations are the same. Only the capture tip will be described.
See FIG.
FIG. 8 is a schematic plan view of a capture chip 50 used in the cell supply / discharge device of Embodiment 2 of the present invention, and an inflow portion is formed at the center of one end of the silicon substrate 51 using a normal photolithography process. 52, an outflow portion 56 is formed at the center of the other end, and a plurality of parallel flow channels 54 are formed between the inflow portion 52 and the outflow portion 56. In addition to being connected by the branch flow path 53, the plurality of flow paths 54 and the outflow portion 56 are connected by the merge flow path 55.

  In addition, a suction through-hole 57 is provided at the bottom of the channel 54, and an opening 58 connected to the through-hole 57 is formed on the back surface of the silicon substrate 51, and cells are sucked from the through-hole 57 through the opening 58. To do.

  Although illustration is omitted, in this case as well, the side wall and the bottom surface of the flow path 44 are coated with a hydrophilic resin, for example, HydroCell (trade name, manufactured by Cellseed) to perform hydrophilic treatment, and silicon other than the flow path 33 is used. The surface of the substrate 31 is coated with a hydrophobic resin, for example, an acrylic resin and subjected to a hydrophobic treatment.

Also in this case, the width of the flow path 54 depends on the size of the cells to be captured, but when the cell size is about 10 to 15 μm, for example, the width is 50 μm and the depth is 50 μm. The thickness is about 500 μm.
The through holes 57 provided in the bottom of the flow channel 54 have a diameter of, for example, 2 μm, and the through holes 57 are arranged at an interval of, for example, 50 μm.

  Thus, in Example 2 of the present invention, since the flow path 54 is branched into a plurality of channels, the transport of the cells 15 can be processed in parallel, and the recovery efficiency can be improved.

  Next, referring to FIG. 9, a microinjection device using the cell supply / discharge / capture device of Example 1 or 2 will be described, except that an injection device 60 is provided at a position facing the capture chip. This is the same configuration as the cell supply / discharge / capture device of Example 1 or 2.

FIG. 9 is a conceptual configuration diagram of the microinjection apparatus according to the third embodiment of the present invention, which includes a pipe suspension 12 and a cell suspension 13 in which cells 15 are suspended from a capillary 12 in a medium 14. Cell supply device 11 for supplying the trapping chip 30 and a pipette-like capillary tube 17 and supplying a carrier liquid 18 having a heavier specific gravity than the culture medium from the capillary tube 17 to the capture chip 30, and the supplied cell suspension. A capture chip 30 that sucks and captures the cells 15 in the liquid 13 by the suction device 23, a container 19 that contains the capture chip and has an opening 20 for suction, holds the container 19, and is connected to the suction device 22 A chip holder 21, a suction device 23 for capturing the cells 15, a cell observation device 40 for observing the captured cells 15, and a captured cell 15 An injection device 60 for injecting a drug solution or a gene, an unnecessary cell recovery device 24 for recovering uncaptured cells 15 through a thin tube 25, and a substance-injected cell subjected to injection work are recovered through a thin tube 27. A basic configuration is configured from the captured cell recovery device 26.

  The injection device 60 in this case includes a substance introduction needle 61 and a substance introduction needle control device 62 that controls the substance introduction needle 61, and the cells 15 captured in the through holes provided in the capture chip 30. On the other hand, substances such as genes or chemicals are sequentially introduced by the substance introduction needle 51.

  Thus, in Example 3 of the present invention, since the microinjection device is configured using the cell supply / discharge device of Example 1 or 2, the collection of the substance-introduced cells is used.

  Although the embodiments of the present invention have been described above, the present invention is not limited to the conditions and configurations described in the embodiments, and various modifications are possible. For example, the capture described in the embodiments The shape, width, depth, and length of the channel provided in the chip are arbitrary, and may be appropriately determined according to the size of the cell together with the diameter of the through hole.

  Further, in each of the above embodiments, an acrylic resin is used for the hydrophobic treatment. However, the resin is not limited to an acrylic resin, and a normal resin has a hydrophobic property. It may be used after confirming whether or not it has hydrophobicity. Furthermore, since silicon itself has hydrophobicity, hydrophobic treatment is not essential.

  In each of the above embodiments, the side walls and bottom surface of the flow path are made hydrophilic so that the cell suspension or liquid can easily flow only in the flow path in the open state, and the surface of the silicon substrate other than the flow path is made hydrophobic. However, such hydrophilic treatment / hydrophobic treatment is not essential.

  For example, the side wall and bottom surface of the flow path may be made hydrophilic and the surface of the silicon substrate other than the flow path may be left as it is, or the surface of the silicon substrate other than the flow path may be made hydrophobic, The side wall and the bottom surface of the flow path may be left as they are, and further, no treatment may be performed on any one of them.

  In each of the above embodiments, the capture chip is a silicon substrate that has been established with microfabrication technology in the field of semiconductor integrated circuits and micromachine technology, but is not limited to a silicon substrate. An acrylic resin may be used.

  Further, in Example 3 described above, a single substance introduction needle 61 is provided in the injection device 60 to inject a gene or a chemical solution or the like sequentially into the cell. The number of substance introduction needles corresponding to the number of the substance introduction needles may be provided in a one-dimensional array, whereby the injection efficiency can be increased.

Further, in each of the above embodiments, a pipette-like thin tube is connected to the cell supply device, and the cell suspension is dropped into the inflow portion. Alternatively, the cell suspension may be connected to the capture chip via the cell supply device.
Such a configuration is the same for the solution supply device in the liquid delivery device.

  In each of the above embodiments, the container for storing the capture chip is made of polystyrene of the same material as the petri dish, but is not limited to polystyrene, and is a chemical resistant solution such as polycarbonate, acrylic resin, or glass. What is necessary is just to comprise with a highly specific substance.

  In Example 1 described above, cells are captured by observing cell division with a through-hole provided in the flow path. However, the present invention is not limited to such a case. The present invention is also applied to the case of observing a chemical reaction of a cell by flowing a chemical solution through a flow path in a state where the provided cell is captured.

The detailed features of the present invention will be described again with reference to FIG. 1 again.
See Figure 1 again
(Supplementary note 1) A cell supply mechanism 1 for supplying a cell suspension 2, a liquid supply mechanism 3 for supplying a liquid 4 not containing cells, a flow path 6 for flowing the cell suspension 2 and the liquid 4, and the cell suspension Capture chip 5 having a through-hole 7 that captures cells in the liquid 2, a suction mechanism 8 that is connected to the capture chip 5 and controls the amount of suction from the through-hole 7, and was not captured by the through-hole 7 A cell supply / drainage / capture apparatus comprising at least an unnecessary cell recovery mechanism 9 for recovering unnecessary cells and a capture cell recovery mechanism 10 for recovering captured cells.
(Additional remark 2) The said capture | acquisition chip | tip 5 has an inflow part which flows the said cell suspension 2 and the liquid 4 to the said flow path 6 in the one end part of the said flow path 6, and the other end part of the said flow path 6. A plurality of through holes 7 having a diameter through which the cells in the cell suspension 2 cannot pass. The cell supply / discharge / capture device according to appendix 1, wherein the device is provided along the flow path 6.
(Supplementary note 3) The cell supply / discharge / capture device according to Supplementary note 2, wherein the flow passage 6 comprises a single meandering flow passage 6.
(Additional remark 4) While the said flow path 6 consists of the several flow path 6 arrange | positioned in parallel, it has the connection flow path 6 connected to a common inflow part at the end of each flow path 6, and each said flow path 6 The cell supply / discharge / capture device according to appendix 2, characterized in that it has a connection channel 6 connected to a common recovery section at the other end of the cell.
(Additional remark 5) While making the inner wall of the said flow path 6 provided in the said capture chip 5 more hydrophilic than the base material of the said capture chip 5, surfaces other than the inner wall of the flow path 6 of the said capture chip 5 are made from the said base material. 5. The cell supply / discharge / capture device according to any one of appendices 1 to 4, wherein the device is hydrophobic.
(Appendix 6) The inner wall of the flow channel 6 is coated with a hydrophilic resin from the base material of the capture chip 5, and the surface other than the inner wall of the flow path 6 of the capture chip 5 is more hydrophobic than the base material. 6. The cell supply / discharge / capture device according to appendix 5, which is coated with
(Supplementary note 7) The cell supply / discharge / capture device according to any one of supplementary notes 1 to 6, wherein the liquid 4 containing no cells is a liquid 4 having a higher specific gravity than the cells.
(Supplementary note 8) The supplementary note 8, wherein the liquid 4 not containing cells is a liquid 4 having the same components as the medium constituting the cell suspension 2 and having a higher salt concentration than the medium. Cell supply / capture device.
(Supplementary note 9) The cell supply / discharge / capture device according to any one of supplementary notes 1 to 8, further comprising an injection mechanism for injecting a substance into the captured cells.
(Supplementary Note 10) A step of flowing the cell suspension 2 from one end of a flow path 6 having a plurality of through-holes 7 and capturing the cells in the cell suspension 2 by suction through the through-holes 7, the flow path 6 Either a step of flowing a liquid 4 having a higher specific gravity than the cell from one end of the cell to float the cell that could not be captured and collecting it at the other end of the flow path 6, an injection for injecting a substance into the captured cell, or a follow-up observation And a step of releasing suction after the injection or follow-up, and flowing a liquid 4 having a higher specific gravity than the cell from one end of the channel 6 to float the cell and recovering the other end of the channel 6 A method for supplying and discharging cells, which is characterized by sequentially performing steps.

  As an example of utilizing the present invention, gene or drug solution is typically introduced into cells in business such as regenerative medicine and genome drug discovery. The present invention is also applied to the case of observing a chemical reaction by flowing it.

It is explanatory drawing of the fundamental structure of this invention. It is a notional block diagram of the cell supply / discharge / capture device of Example 1 of the present invention. It is a schematic plan view of the capture chip of Example 1 of the present invention. It is a schematic sectional drawing of the acquisition chip of Example 1 of the present invention. It is explanatory drawing of the arrangement structure of the capture | acquisition chip | tip thru | or chip | tip holder of Example 1 of this invention. It is an operation | movement flowchart of the cell supply / discharge / capture apparatus of Example 1 of this invention. It is explanatory drawing of the capture | acquisition and conveyance of the cell in Example 1 of this invention, and an open | release and conveyance state. It is a schematic plan view of the capture | acquisition chip | tip used for the cell supply / discharge | capture apparatus of Example 2 of this invention. It is a notional block diagram of the microinjection apparatus of Example 3 of this invention.

Explanation of symbols

DESCRIPTION OF SYMBOLS 1 Cell supply mechanism 2 Cell suspension 3 Liquid supply mechanism 4 Liquid 5 Capture chip 6 Channel 7 Through-hole 8 Suction mechanism 9 Unnecessary cell recovery mechanism 10 Captured cell recovery mechanism 11 Cell supply device 12 Capillary tube 13 Cell suspension 14 Medium 15 Cell 16 Liquid feeder 17 Narrow tube 18 Carrier liquid 19 Container 20 Opening portion 21 Chip holder 22 Connection portion 23 Suction device 24 Unnecessary cell collection device 25 Narrow tube 26 Captured cell collection device 27 Narrow tube 28 Connection member 30 Capture chip 31 Silicon substrate 32 Inflow portion 33 Channel 34 Outflow portion 35 Through hole 36 Opening portion 37 Hydrophilic resin 38 Hydrophobic resin 40 Cell observation device 41 Light source 42 Objective lens 43 Image processing device 50 Capture chip 51 Silicon substrate 52 Inflow portion 53 Branching channel 54 flow path 55 merge flow path 56 outflow part 57 through hole 58 opening part 60 injection device 61 substance introduction needle 62 substance Needful needle control device

Claims (5)

A cell supply mechanism for supplying a cell suspension, a liquid supply mechanism for supplying a liquid not containing cells, a flow path for flowing the cell suspension and liquid, and a through-hole for capturing cells in the cell suspension. A trapping chip, a suction mechanism that is connected to the trapping chip and controls the amount of suction from the through-hole, an unnecessary cell recovery mechanism that recovers unnecessary cells that are not trapped in the through-hole, and a trapped cell is recovered A cell supply / discharge / capture device comprising at least a capture cell recovery mechanism. The capture chip has an inflow portion for flowing the cell suspension and liquid in the flow channel at one end of the flow channel, and the cell suspension from the flow channel to the other end of the flow channel. And a recovery part for recovering the liquid, and the through-hole is formed of a plurality of through-holes having a diameter through which cells in the cell suspension cannot pass, and is provided along the flow path The cell supply / discharge / capture device according to claim 1. The inner wall of the flow path provided in the capture chip is made more hydrophilic than the base material of the capture chip, and the surface other than the inner wall of the flow path of the capture chip is made more hydrophobic than the base material. The cell supply / discharge / capture device according to claim 1 or 2. 4. The cell supply / discharge / capture device according to claim 1, wherein the liquid containing no cells is a liquid having a specific gravity higher than that of the cells. 5. A step of flowing a cell suspension from one end of a flow path provided with a plurality of through-holes and capturing the cells in the cell suspension by suction through the through-holes; a liquid having a higher specific gravity than the cells from one end of the flow path A step of floating and collecting cells that could not be captured at the other end of the flow path, a step of injecting a substance into the captured cells or a follow-up observation, aspiration after the injection or follow-up observation And supplying the cell supply / discharge / capture characterized by sequentially performing a step of releasing a liquid having a higher specific gravity than the cell from one end of the flow path to float the cell and collecting it at the other end of the flow path Method.

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