JP2007222132A - Cell capture chip - Google Patents

Abstract

The present invention relates to a cell capture chip that individually captures cells of animals and the like, and is capable of realizing cell capture that is excellent in cell fixation and has a small load on the cells accompanying cell fixation, and is capable of performing injection treatment by a microinjection method. Enables mass processing.
A cell trapping chip (1) has one or more cell trapping portions (3) for trapping one cell (2), and a suction hole (4) provided for each cell trapping portion (3), and cells existing in a liquid 2 is a cell trapping chip 1 that traps the cell trapping part 3 in the cell trapping part 3 by negative pressure from the suction hole 4, and the cell trapping part 3 has an opening size through which one cell can pass by the negative pressure or positive pressure from the suction hole 4. A lid portion 5 having a cell inlet / outlet 6 is provided, and the inside of the cell capturing portion closed by the lid portion 5 is a microwell that can accommodate only one cell 2.
[Selection] Figure 1

Description

  The present invention relates to a cell capture chip that individually captures cells of animals and the like, and more particularly to a cell capture chip that continues to capture cells without being attracted or adsorbed by the cells.

  In recent years, modification of cell properties by injecting genes or the like into cells has attracted attention as a method for treating diseases caused by genetic causes, and research is progressing. The results of this research include elucidation of the role of genes and the possibility of tailor-made medicine that provides medications and treatments tailored to individual genetic characteristics.

  Methods for injecting genes into cells include electrical methods using high-pressure pulses (electroporation method), chemical methods using endocytosis and membrane fusion (lipofection method), and organisms using viruses. Generally known methods (virus vector method), mechanical methods using microneedles (microinjection method), optical methods using laser light (laser injection method) and the like are well known. However, the electrical method has a large damage to the cells, and the chemical method has poor gene injection efficiency. In addition, biological methods have disadvantages such as not being able to introduce all of the injected material (virus limitation). On the other hand, at the present time, the mechanical method is attracting attention as the safest and most efficient injection method.

As shown in FIG. 7, a conventional mechanical method includes a cell fixing plate (generally a Si chip) having a plurality of holes smaller than the cell diameter for capturing cells, and a culture solution containing cells. By sucking (liquid medium) from the suction hole provided in the lower part of the hole, the cells are adsorbed and fixed to the surface portion of the hole on the cell fixing plate. Although not shown in the figure for fixing cells, other means are those that apply a high frequency between the electrodes and fix the cells to the microelectrodes on the cell fixation plate using the force of the electric field acting on the cells, or affinity for the cells For example, a cell-affinity substance is placed on a cell-fixing plate having no cell, and cells are fixed on the cell-affinity substance. There is a method of injecting (injecting) a substance by inserting a fine injection needle (capillary) into a cell fixed by any one of these means (see, for example, Patent Document 1). In this method, a large amount of injection processing into cells is performed by sequentially injecting a substance with an injection needle into each of a plurality of cells adsorbed and fixed on a cell fixing plate.
JP-A-1-112976

  However, in the case of fixing cells by suction, which is a conventional technique, from the time when the cells are captured until the substance is completely injected into the cells with the injection needle, the cells are prevented from detaching and moving from the cell fixing plate. Since the cells must be continuously sucked from the suction hole, there is a problem that the cell wall in contact with the suction hole is always in a negative pressure state and the cell receives stress or damage. In addition, it is possible to increase the volume of the injection process by increasing the number of holes on the cell fixation plate, but the total time until the injection process to all cells is completed as the number of cells to be processed increases. Therefore, there is a problem that it is unsuitable for further mass processing due to an increase in stress and damage to the cells that are continuously given during that time.

  In other cases where cells are fixed by applying high frequency, if the application of high frequency is reduced in consideration of stress or damage to the cells, the force to fix the cells will be weakened, and the cells will be fixed by cell affinity substances. In the case of fixing the cells, although the stress and damage to the cells are small, it is difficult to obtain a sufficient force to fix the cells. Therefore, both of them have a problem that the position stability of the cells when the injection needle is inserted into the cells is lacking. is there.

  In view of the above, there is a need for a cell trapping means that is excellent in cell fixation during cell trapping, has low stress and damage to the cells, and is suitable for large-scale injection processing by the microinjection method.

  Accordingly, the present invention has an object to provide a cell trapping chip that can achieve cell trapping that is excellent in cell fixation and has a small load on the cells accompanying cell fixation, and that enables mass processing of injection processing by the microinjection method. And

  The cell trapping chip of the first invention has one or more cell trapping portions for trapping one cell, and a suction hole provided for each of the cell trapping portions, and sucks out the cells present in the liquid. A cell capturing chip that captures in a cell capturing part by negative pressure from the lid, wherein the cell capturing part has a cell inlet / outlet having an opening size through which one cell can pass by negative pressure or positive pressure from the suction hole And the inside of the cell trapping part closed by the lid part is configured as a cell accommodating space that can accommodate only one cell.

  The cell trapping chip of the second invention is the cell trapping chip of the first invention, wherein the cell entrance / exit has a spherical diameter inscribed in the cell entrance / exit when the average cell diameter of the cell group is 1. The configuration is within the range of 0.2 or more and less than 1.0.

  The cell trapping chip of the third invention is the cell trapping chip of the first invention, wherein the cell housing space has a spherical diameter inscribed in the cell housing space when the average cell diameter of the cell group is 1. It is set as the structure which exists in the range of 1.0 or more and less than 2.0.

  A cell capture chip according to a fourth aspect of the present invention is the cell capture chip according to the first aspect, wherein the cell entrance / exit has one or more protrusions substantially perpendicular to the cell passage direction. To do.

  The cell trapping chip of the fifth invention is the cell trapping chip of the first invention, wherein the lid portion has one through-hole having an opening size in which one cell cannot pass by negative pressure or positive pressure from the suction hole. The above configuration is further included.

  According to the first, second, and third inventions, the cell capturing unit is provided with the lid portion having the cell entrance / exit having an opening size through which one cell can pass by the negative pressure or the positive pressure from the suction hole. Since the inside of the closed cell capturing part is a cell accommodating space that can accommodate only one cell, after passing the cell inlet / outlet by negative pressure from the suction hole and capturing one cell in the cell capturing part, The cell can be securely held in the cell trapping portion by the lid portion and can be fixed.

  Further, according to the fourth and fifth inventions, since the lid portion has a space (including a through hole) at a location different from the cell entrance / exit, the cells captured in the cell capturing portion are removed from the space. Observation and manipulation can be performed from a different angle from the cell entry / exit.

  According to the present invention, one cell that has passed through the cell inlet / outlet of the lid and trapped in the cell trapping part is securely fixed in the cell trapping part by the lid, regardless of external force to the cells. Therefore, it is possible to achieve cell trapping that is excellent in cell fixation and has a low load such as stress and damage to the cells accompanying cell fixation.

  In addition, since the lid has a space such as a through hole at a location different from the cell entry / exit, the cells captured in the cell capture unit can be manipulated or observed from the space.

  Furthermore, since the cells trapped in the cell trapping part can be released from the cell inlet / outlet by the positive pressure from the suction hole, the cells that have been subjected to the injection treatment can be easily recovered.

  Therefore, since it is possible to realize a large amount of injection processing by the microinjection method, it is possible to reduce processing time and cost associated with the injection processing.

(Example 1)
Hereinafter, embodiments of the present invention will be described with reference to the drawings. FIG. 1 is a diagram showing the structure of a cell trapping chip according to an embodiment of the present invention. In FIG. 1, (a) is a cross-sectional view of the cell trapping chip, and (b) is an enlarged view of a cell trapping portion. The perspective view and the diagram (c) in the figure show the cell trapping state. For convenience of explanation, the cell trapping part is described here as a shape having a hemispherical cell storage space, and the opening shape of the cell entry / exit is described as a circle.

  In FIG. 1 (a), the cell trapping chip 1 is provided with one or more cell trapping sections 3 that trap one cell 2. In FIG. 1 (b), the cell trapping part 3 has a hemispherical microwell structure, has a suction hole 4 for applying a negative pressure or a positive pressure in the cell trapping part 3, and has a cell trapping part. 3 is provided with a lid portion 5 having a cell inlet / outlet 6 having an opening size through which one cell can pass by negative pressure or positive pressure from the suction hole 4. The cell trapping part 3 closed by the lid part 5 has a structure having a cell accommodating space 7 in which only one cell 2 can be accommodated. In FIG. 1 (c), one cell 2 existing in a cell accommodating space (hereinafter referred to as a microwell) 7 in the cell trapping portion 3 is a cell 2 in a trapped state.

  As for the installation position of the cell entrance / exit 6, it is more desirable that the center of the opening shape is relatively located at the center of the microwell 7. The same applies to the installation position of the suction hole 4. That is, when the cell inlet / outlet 6 and the suction hole 4 are circular, the center thereof is a position corresponding to the center of the hemisphere in the hemispherical microwell.

  The opening size of the cell entrance / exit 6 is set so that the spherical diameter inscribed in the cell entrance / exit 6 is 0.2 or more and less than 1.0 when the average cell diameter of the cell group to be captured is 1. In the state where there is no negative pressure or positive pressure from the suction hole 4, the cell cannot pass through the cell inlet / outlet 6, whereas when the cell 2 passes through the cell inlet / outlet 6, excessive stress is applied to the cell 2. It is preferable to set it to a size that does not exist. For example, when the cell diameter is 20 μm, the circular diameter of the cell inlet / outlet 6 is in the range from 4 μm to less than 20 μm, and preferably about 10 μm.

  Similarly, the opening size of the suction hole 4 is set within a range of less than 0.2, and the cell 2 captured in the microwell 7 is set to a size that does not pass through the suction hole 4 due to the negative pressure of the suction hole 4. The negative pressure value of the suction hole 4 when each size is within the above range is preferably about 450 Pa, for example.

  Further, the size of the microwell 7 is set so that the spherical diameter inscribed in the microwell 7 is 1.0 or more and less than 2.0 when the average cell diameter is set to 1. It is preferable to set the cell 2 accommodated in the cell 2 so that stress due to the pressure from the wall of the microwell 7 is not applied and the free movement distance of the cell 2 is shortened.

  Subsequently, an operation of capturing the cell 2 by the cell capturing chip 1 will be described. The cell suspension, which is a liquid containing the cells 2 filled on the cell trapping chip 1, receives negative pressure suction at 450 Pa from below through the suction holes 4. It is attracted to the microwell 7.

  When the cell diameter of the cell group to be captured is about 20 μm, for example, when the diameter of the cell inlet / outlet 6 is 10 μm (opening size of 0.5 with respect to the cell diameter of 20 μm), negative pressure suction from the suction hole 4 Accordingly, the cells 2 of about 20 μm pass through the cell inlet / outlet 6 and are introduced into the microwell 7 and captured (accommodated). However, when a cell 2 of about 100 μm is included in the cell group, the cell 2 cannot pass through the cell inlet / outlet 6 having a diameter of 10 μm (an opening size of 0.1 with respect to the cell diameter of 100 μm). For example, the opening portion of the outlet 6 is not blocked and introduced into the microwell 7. From this, it becomes possible to reliably capture only the cells 2 having substantially the same size, that is, the cells 2 to be captured, in the microwell 7.

  The cells 2 once captured in the microwells 7 cannot pass through the cell inlet / outlet 6 even if the negative pressure suction is stopped because the diameter of the cell inlet / outlet 6 is smaller than the cell diameter. Since it is fastened, it does not float outside the microwell 7. Therefore, after the cell 2 is captured, the negative pressure suction from the suction hole 4 can be stopped, and the stress on the cell 2 due to the negative pressure suction can be eliminated. This makes it possible to extremely reduce damage to the cells 2 during the injection process, so that it is possible to accurately observe the reaction and function expression of the cells 2 with respect to the injection substance (chemical solution or the like). Note that if the plurality of suction holes 4 are divided into blocks and negative pressure suction is performed in units of blocks, the stress on each cell 2 due to negative pressure suction during the injection process can be further reduced.

  In addition, since the cell 2 captured in the microwell 7 has a short movement free distance, when a microinjection needle (capillary) is inserted into the cell 2 from the cell inlet / outlet 6 outside the microwell 7, Since the cell 2 does not largely escape from the needle tip, a substance (chemical solution or the like) can be easily injected.

  In addition, after the injection process to the cells 2 is completed, by applying a positive pressure through the suction hole 4, the captured cells 2 pass through the cell inlet / outlet 6 and outside the microwell 7, that is, Since it can be detached from the cell trapping chip 1, all of the cells 2 after the injection treatment can be collected. The collected cells 2 are used for post-process reaction detection and culture, which are not described here.

  Next, a manufacturing process of the cell trapping chip 1 will be described with reference to the drawings. FIG. 2 is a diagram showing a process for manufacturing a cell trapping chip, and (a) to (e) in FIG. 2 correspond to (a) to (e) of the following explanation items. Since the cell trapping chip 1 is generally convenient to manufacture using an SOI (Silicon On Insulator) substrate because of its manufacturability and the like, an example using an SOI substrate will be described.

  (a) First, after cleaning the SOI substrate, a photoresist for patterning the device layer (Si layer) is applied. The material and thickness of the photoresist used here are not particularly limited. Next, the photoresist is exposed using a photomask and then developed. For the exposure of the photoresist, a mask aligner generally used in semiconductor manufacturing is used. The patterned resist is used as an etching mask for the Si through hole (suction hole 4). A through-hole is formed by using a dry etching apparatus for forming the Si through-hole.

(b) Next, in order to produce a hemispherical microwell shape in the device layer (microwell formation), an etching mask is produced on the surface of the device layer. When wet etching is used in the next step, it is preferable that the etching mask covers the entire SOI substrate with a thermal oxide film. When dry etching is used, the etching mask may be a photoresist, SiO ₂ (silicon oxide) or Si ₃ N ₄ (silicon nitride) formed by sputtering or CVD (Chemical Vapor Deposition). .

In the case of a photoresist as an etching mask in wet etching and dry etching, patterning by exposure / development is performed using a photomask in the same manner as in the above (a). On the other hand, in the case of SiO _2, the mask for patterning the SiO ₂ is required. A photoresist is used for patterning of SiO _{2, and} the patterned photoresist opening is patterned with an HF (hydrofluoric acid) aqueous solution.

  After producing the etching mask, the silicon of the device layer is etched to form a microwell. In order to perform hemispherical etching as shown in FIG. 2, it is desirable to use isotropic etching. Isotropic etching of silicon is possible by changing dry etching conditions (for example, gas flow rate ratio).

  The cell accommodation space 7 of the hemispherical microwell can accommodate only one cell 2 (a space where two cells cannot be accommodated), and the cell diameter of the cell group is 1 so that the cell 2 does not move freely during the injection process. In this case, the spherical diameter inscribed in the cell accommodating space 7 is preferably in the range of 1.0 or more and less than 2.0.

(c) After producing the microwell shape, the process proceeds to the production process of the lid 5. The lid 5 is formed using a material having transparency and a certain degree of hardness (for example, moderate hardness that does not easily damage cells and does not easily break), for example, SiO ₂ or epoxy resin SU-8. . When the SiO ₂ film is formed by the CVD method or the like, it is also formed on the surface of the cell trapping part 3, so that the entire cell trapping part 3 needs to be filled with a sacrificial layer or the like (for example, a resist). On the other hand, the same applies when using SU-8 resist (thick film resist manufactured by US Microchem Co., Ltd.).
By overcoating thick film resist, the entire microwell structure is filled with resist.

  Next, after baking the resist, the surface of the device layer and the resist sacrificial layer are polished for planarization. Polishing uses a CMP (Chemical Mechanical polishing) method which is one of mechanical polishing methods. By using the CMP method, both the resist sacrificial layer and the device layer become flat surfaces having a uniform height.

(d) After flattening by the CMP method, the lid 5 is formed. For SiO ₂ , a transparent SiO ₂ film can be produced by using a liquid source CVD method using an organic silicon-based material such as TEOS (tetraethoxysilane). A resist is applied again on the surface of the SiO ₂ film of about 20 μm produced by the CVD method, and exposure and development are performed through a photomask (patterning mask). Subsequently, SiO ₂ patterning is performed by dry etching or the like. Then, the cover part 5 is produced by removing the resist sacrificial layer by plasma ashing.

  Moreover, SU-8 can produce a thick film-shaped transparent structure by a conventional spin coating method. When SU-8 is used, the lid 5 can be formed in a shape that is only partially opened by exposure and development. Furthermore, the resist sacrificial layer can be removed by using a resist stripping solution.

  The resist sacrificial layer is not particularly limited as long as it is a material that can be formed into a thick film and has heat resistance and can be removed by a resist stripping solution or plasma etching.

(e) Subsequently, the SOI substrate layer is etched. The substrate layer needs to have a certain height as a support for the cell trapping chip 1. In the etching of the substrate layer, it is preferable to use SiO ₂ for the etching mask because of the etching selectivity between Si and the etching mask. Therefore, in order to cover the silicon of the device layer, SiO ₂ is formed again on the entire SOI surface by thermal oxidation. Of course, a material other than SiO ₂ , for example, Si ₃ N ₄ may be used.

In order to pattern the SOI back surface SiO ₂ , a photoresist is applied to the SOI back surface, and development and exposure are performed. After patterning SiO ₂ with HF or the like, and then anisotropically etching the substrate layer silicon from the back surface with an alkaline aqueous solution or the like, and removing the SOI insulating film, the shape as shown in FIG. The Note that dry etching or the like may be used for silicon etching of the SOI substrate layer.

The manufacturing process, material, shape, dimensions, and the like described above are not limited to the contents described here as long as the cell trapping chip 1 having the same function and structure as the present invention can be manufactured. For example, the shape of the microwell may be an elliptical sphere similar to a hemispherical shape, and the opening shape of the cell inlet / outlet 6 is an elliptical shape, a triangular shape, a quadrangular shape, a polygonal shape, or a deformation thereof. It doesn't matter. In the above-described step (c), the lid portion 5 is made of a material having transparency if possible. The reason is that the cells 2 trapped in the microwells 7 are transmitted through the lid portion 5 to be observed with a microscope or the like. This is because it is convenient for observation.
(Example 2)
The present invention is the cell trapping chip 1 in which the cell-accommodating space 7 in the cell trapping chip 1 described in Example 1 is a microwell having a shape (a prismatic shape, a columnar shape, a pyramid shape, etc.) different from a hemispherical shape. . FIG. 3 is a diagram (an enlarged perspective view of a cell trapping portion) showing a structure of a cell trapping chip according to another embodiment of the present invention. Below, the manufacturing process of the cell capture chip | tip 1 of this invention is demonstrated.

  3 (a) and 3 (b) show a microwell structure having a cylindrical shape and a rectangular parallelepiped shape. The process up to the fabrication of the microwell of this shape is the same as that in the first embodiment (the process (a) described above), and the process after the fabrication of the microwell is the same as that in the first embodiment (the above (c) to (e). Process). That is, only the microwell formation method (step (b) described above) in FIG. 2B, which is a process between them, is different, and only this portion of the process will be described below.

  After the SOI surface device layer is etched by the process (a) described above, an etching mask for forming a microwell structure is patterned. The etching mask is circular when the microwell structure is cylindrical, and is square (or rectangular, etc.) when it is a rectangular parallelepiped. Then, by vertically etching the device layer silicon by dry etching or the like, a cylindrical or rectangular parallelepiped microwell as shown in FIGS. 3A and 3B is manufactured.

  On the other hand, anisotropic etching such as silicon etchant (EPW (Ethylenediamine Pyrochatechol Water) or TMAH (Tetra Methyl Ammonium Hydroxide), KOH (potassium hydroxide aqueous solution), etc.) is used for etching. In this case, a pyramid-shaped microwell as shown in FIGS. 4C and 4D can be produced. When the above etchant is used, the etched surface exhibits a silicon crystal plane anisotropy and becomes a tapered shape, and the shapes as shown in FIGS. 4C and 4D can be obtained depending on the etching time.

  Here, the cell accommodation space 7 of the microwell here can also accommodate only one cell 2 (a space in which two cells cannot be accommodated) as in the case of the hemispherical microwell, and the cell 2 does not move freely during the injection process. As described above, when the average cell diameter of the cell group is 1, it is desirable that the spherical diameter inscribed in the cell accommodating space 7 is in the range of 1.0 or more and less than 2.0.

In addition, the above-described manufacturing process, material, shape, size, and the like are not limited to the contents described here as long as the cell trapping chip 1 having the same function and structure as the present invention can be manufactured. For example, the shape of the microwell may be a cylindrical shape, a prismatic shape, a conical shape, a pyramid shape, or a modification thereof.
Example 3
The present invention is different from the cell trapping chip 1 described in the first embodiment in that the opening shape of the cell inlet / outlet 6 in the lid portion 5 is circular, and one or more protrusions substantially perpendicular to the cell passage direction. 1 is an opening-shaped cell trapping chip 1 having a portion. FIG. 4 is a view showing another shape (eave structure) of the cell entrance / exit of the present invention, and FIG. 5 is a view showing another modified shape of the cell entrance / exit of the present invention.

  The process for producing the cell trapping chip 1 of the present invention will be described below. First, the shape of the cell inlet / outlet 6 is four convex portions (hereinafter referred to as eaves structure) as shown in FIG. What is formed will be described.

The process steps of the present invention are the same as those of the first embodiment described above except for the patterning for the lid 5. The photomask of the lid 5 (patterned mask) prepared as in FIG. 4 (a), by performing exposure and development of the SiO ₂ etching or SU-8, eaves structure as shown in FIG. 4 (b) Is produced. In addition, it is possible to similarly manufacture a product formed of four convex portions as shown in FIG.

Next, what forms two convex parts (eave structure) by overlapping two circular shapes as shown in FIG. 5 (a) will be described. The process steps of the present invention are the same as those in the first embodiment described above except for the patterning for the lid 5. A photomask for the lid 5 is produced as shown in FIG. 5 (a), and the eaves structure as shown in FIG. 5 (b) is produced by etching SiO ₂ or exposing and developing SU-8. .

  The cell entrance / exit 6 of the present invention is the only part through which the cell 2 can pass through the lid 5 and the opening size is a spherical shape inscribed in the cell entrance / exit 6 when the average cell diameter of the cell group is 1. It is desirable that the diameter is within the range of 0.2 or more and less than 1.0.

  In addition, the convex portion of the cell entrance / exit 6 is moderate so as not to damage or damage the cell 2 when the cell 2 enters or exits due to a part thereof, or to apply unnecessary pressure stress to a part of the cell 2. It is preferable to have a curved surface.

  The manufacturing process, material, shape, dimensions, and the like described above are not limited to the contents described here as long as the cell trapping chip 1 having the same function and structure as the present invention can be manufactured. For example, the shape of the protrusions does not need to be similar to each other, and the opening space of the lid 5 need not be point-symmetric or line-symmetric.

With the lid 5 according to the present invention, an appropriate space (such as a gap) is obtained between the convex portions by devising the shape of the convex portion (eave structure), and the cells 2 can be observed from the space with a microscope or the like. In addition, it is possible to inject the cells 2 with a fine injection needle. In addition, depending on the form of the space between the convex portions, it is possible to perform an injection process from other spaces to the cells with a fine injection needle while performing observation with a microscope.
Example 4
The present invention is the cell trapping chip 1 having a through hole which is one or more openings different from the cell inlet / outlet 6 in the lid portion 5 in the cell trapping chip 1 described in the first embodiment. FIG. 6 is a view showing a lid portion having a plurality of through holes according to the present invention.

  Below, the manufacturing process of the cell capture chip | tip 1 of this invention is demonstrated.

The process steps of the present invention are the same as in Example 3 described above. A photomask (patterning mask) for the lid 5 is produced as shown in FIG. 6, and SiO ₂ etching or SU-8 exposure / development is carried out to produce the eaves structure having the through hole shown in FIG.

  The opening size of the through hole of the present invention is such that the cell 2 cannot pass therethrough, and when the average cell diameter of the cell group is 1, the spherical diameter inscribed in the through hole may be less than 0.2. desirable.

  The manufacturing process, material, shape, dimensions, and the like described above are not limited to the contents described here as long as the cell trapping chip 1 having the same function and structure as the present invention can be manufactured. For example, the opening shape of the through hole may be a circle, an ellipse, a triangle, a rectangle, a polygon, a star, or the like. In addition, the inner wall surface of the through hole may not be perpendicular to the surface of the lid 5, and may be inclined toward the vicinity of the center of the cell 2 captured in the microwell, for example.

  Since the lid portion 5 according to the present invention has a through hole having an opening different from that of the cell inlet / outlet 6, it is possible to perform an injection process from the through hole to the cell 2 with a fine injection needle, for example, when a plurality of through holes are provided. (FIG. 6), the continuous injection | pouring process using a some fine injection needle from each through-hole is attained.

  In addition, since the cells 2 can be observed from the through hole with a microscope or the like as in Example 3, the injection process into the cells from the other through holes with a fine injection needle can be performed while observing with the microscope from the through hole. It becomes possible.

The following notes are disclosed with respect to the above embodiments.
(Appendix 1)
It has one or more cell trapping parts that trap one cell and a suction hole provided for each cell trapping part, and traps the cells present in the liquid in the cell trapping part by negative pressure from the suction hole. A cell capture chip that
The cell trapping part is
A cell having a cell entrance / exit of an opening size through which one cell can pass by negative pressure or positive pressure from the suction hole is provided, and the inside of the cell trapping part closed by the cover can accommodate only one cell. A cell-capturing chip characterized by being a storage space.
(Appendix 2)
The cell entry / exit is
The cell trapping chip according to appendix 1, wherein a spherical diameter inscribed in the cell entrance / exit is within a range of 0.2 or more and less than 1.0, where an average cell diameter of the cell group is 1.
(Appendix 3)
The cell accommodating space is
The cell trapping chip according to appendix 1, wherein a spherical diameter inscribed in the cell accommodation space is 1.0 or more and less than 2.0 when the average cell diameter of the cell group is 1.
(Appendix 4)
The cell entry / exit is
The cell trapping chip according to supplementary note 1, comprising one or more protrusions substantially perpendicular to a cell passing direction.
(Appendix 5)
The lid is
The cell trapping chip according to appendix 1, further comprising one or more through-holes having an opening size through which one cell cannot pass through the negative or positive pressure from the suction hole.
(Appendix 6)
The through hole is
The cell trapping chip according to appendix 5, wherein the inscribed spherical diameter is less than 0.2 when the average cell diameter of the cell group is 1. (Appendix 7)
The lid is
The cell capture chip according to appendix 1, wherein the cell capture chip is made of a material having transparency.

The figure which shows the structure of the cell capture chip | tip of the Example of this invention. The figure which shows the preparation process of the cell capture chip | tip of this invention The figure which shows the structure of the cell capture chip | tip of another Example. Diagram showing other shapes (eave structure) of cell entry / exit Diagram showing other deformed shapes of cell entry / exit The figure which shows the cover part which has several through-holes Conventional cell capture plate

Explanation of symbols

DESCRIPTION OF SYMBOLS 1 Cell capture chip 2 Cell 3 Cell capture part 4 Suction hole 5 Lid part 6 Cell entrance / exit 7 Cell accommodation space

Claims (5)

It has one or more cell trapping parts that trap one cell and a suction hole provided for each cell trapping part, and traps the cells present in the liquid in the cell trapping part by negative pressure from the suction hole. A cell capture chip that
The cell trapping part is
A cell having a cell entrance / exit of an opening size through which one cell can pass by negative pressure or positive pressure from the suction hole is provided, and the inside of the cell trapping part closed by the cover can accommodate only one cell. A cell-capturing chip characterized by being a storage space. The cell entry / exit is
The cell trapping chip according to claim 1, wherein when the average cell diameter of the cell group is 1, the diameter of a sphere inscribed in the cell entrance / exit is in the range of 0.2 or more and less than 1.0. . The cell accommodating space is
2. The cell trapping chip according to claim 1, wherein when the average cell diameter of the cell group is 1, the diameter of a sphere inscribed in the cell accommodation space is in a range of 1.0 or more and less than 2.0. . The cell entry / exit is
The cell trapping chip according to claim 1, comprising one or more protrusions substantially perpendicular to the cell passage direction. The lid is
The cell-capturing chip according to claim 1, further comprising one or more through-holes having an opening size through which one cell cannot pass by negative pressure or positive pressure from the suction hole.

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