Microscopic Object Capturing Device and Microscopic Object Capturing System

Abstract

To provide a microparticle capturing device and a microparticle capturing system that can easily capture microparticles in a sample liquid with a simple structure. 【Solution means】A microparticle capturing device having a chip member and a peripheral wall portion. The chip member has a recess formed on the front side and is provided with a pillar group composed of a plurality of pillars protruding from the surface of the recess. The peripheral wall portion stands on the outer peripheral portion of the surface of the chip member and surrounds the pillar group from at least three directions. The upper end of the peripheral wall portion is higher than the position of the upper end of each pillar. The peripheral wall portion is formed, for example, in a U-shaped (U-shaped with corners right angles) or rectangular frame shape in a plan view. A partition portion is fitted into the U-shaped peripheral wall portion.

Description

Technical Field

[0001] The present invention relates to a microparticle capture device and a microparticle capture system for capturing microparticles in a sample liquid.

Background Art

[0002] In recent years, in the field of life science research, research and development aiming at applications in the food and medical fields have been promoted for microfluidic devices (microfluidic chips) based on microfluidics by devising their structures and the like. Against such a background, there is a need to perform separation and alignment according to size for measurement of various microparticles such as various cells, cell organelles, cell secretions, microorganisms, droplets, microgels, and crystals.

[0003] Cells may have different size and morphology depending on the difference in the division stage (such as G1, S, G2, M phases), and by separating them according to the difference in size, it becomes possible to fractionate them for each division stage. In addition, cells contain cell organelles such as cell nuclei, mitochondria, ribosomes, and Golgi bodies, and the cell organelles differ in size and shape for each type. In particular, the cell nucleus is relatively large compared to other organelles and contains genes, so it is of great significance to separate it when applying genes in biotechnology research.

[0004] Also in recent years, in the fields of medical and immunological research, it has become clear that between tissues such as between cells and between organs, particles smaller than cells called exosomes secrete growth factors and information transfer substances, and when conducting research on such exosomes, there is also an increasing demand to separate cells and exosomes.

[0005] In addition, in the field of regenerative medicine, attempts are being made to produce and transplant sheet-like tissues called cell sheets. However, when culturing starts from a state where cells are sparsely present, when formed into a sheet, the thickness may vary depending on the position, or differences in function may occur. Therefore, there is a desire to evenly align cells at the initial stage of culturing for cell sheet production. Also, in research using microorganisms, there is a desire to separate based on their morphology and size, such as when isolating novel useful microorganisms.

[0006] In the field of biotechnology research, especially in research using droplets, which is rapidly expanding, it would be useful to be able to separate or align single cells based on size in the scenarios of single cell separation and observation. For example, by using a droplet generation chip, when generating w / o droplets (Water-in-Oil droplets: micro water droplets) in an oil phase, the process of simultaneously encapsulating each individual cell can be performed quickly and stably. Culturing cells in a state where one cell is encapsulated in one droplet (a state where one cell is inside one micro water droplet), and confining the substances produced by the cells within the droplet, it is possible to confirm whether each individual cell is producing useful substances. Additionally, if PCR is performed in a state where there are many droplets encapsulating one cell or one virus, it is possible to detect by fluorescence observation in which droplet the cell or virus to be detected was present.

[0007] When aligning and detecting minute objects, it will also be important in the future to be able to keep them stationary on the spot. For example, when it is desired to perform qualitative or quantitative evaluation of the products produced by cells in a droplet, if the droplet moves, it is assumed that stable measurement results cannot be obtained. Especially in measuring instruments (such as spectrometers) that perform scanning, if the measurement position moves, the concentration of the measurement target may change, or the substances constituting the measurement target may change, and reliable data cannot be obtained if it is lost midway.

[0008] Conventionally, when it is desired to separate minute objects, a method (cell sorting) is employed in which minute objects are caused to flow through a microchannel, light is applied to the minute objects in the channel from the outside to perform size measurement and fluorescence detection, and then they are separated electrically (see, for example, Patent Document 1). Further, when it is desired to align minute objects, conventionally, a method is adopted in which a microchamber having a microhole structure or the like is fabricated, and the size is controlled so that one cell falls into one hole, thereby aligning the cells (see, for example, Patent Document 2).

Prior Art Documents

Non-Patent Documents

[0009]

Patent Document 1

Patent Document 2

Summary of the Invention

Problems to be Solved by the Invention

[0010] However, although the conventional method as in Patent Document 1 is suitable for high-speed processing, it is difficult to align minute objects at the same pitch or to make minute objects stationary due to the flow rate performance of existing pumps. On the other hand, according to the conventional method as in Patent Document 2, it is possible to some extent to align cells or make cells stationary, but it is technically difficult to drop cells one by one into the majority of the holes.

[0011] The present invention has been made to solve the above-described problems, and an object thereof is to provide a minute object capturing device and a minute object capturing system that can easily capture minute objects in a sample liquid with a simple structure.

Means for Solving the Problems

[0012] The micro-object capture device according to one aspect of the present invention includes a chip member having a pillar group composed of a plurality of pillars protruding from the surface of a recess formed on the front side, a peripheral wall portion erected on the outer peripheral portion of the surface of the chip member and surrounding the pillar group from three directions, and a partition portion whose both side portions are sandwiched by a pair of opposing side walls of the peripheral wall portion and surrounding the four sides of the pillar group together with the peripheral wall portion. The upper end of the peripheral wall portion and the upper end of the partition portion are higher than the positions of the upper ends of the respective pillars.

[0013] The micro-object capture device according to one aspect of the present invention includes a chip member having a pillar group composed of a plurality of pillars protruding from the surface of a recess formed on the front side, and a peripheral wall portion erected on the outer peripheral portion of the surface of the chip member and surrounding the pillar group from four directions. The upper end of the peripheral wall portion is higher than the positions of the upper ends of the respective pillars.

[0014] The micro-object capture system according to one aspect of the present invention includes the above-described micro-object capture device provided with an injection hole for injecting a sample liquid in at least one of the chip member and the peripheral wall portion, a tubular injection port having one end attached to the injection hole, an injection device including a reservoir that is connected to the injection port and stores the sample liquid, and an electroosmotic flow pump that sends the sample liquid in the reservoir to the recess through the injection port.

Effects of the Invention

[0015] Since the micro-object capture device according to the present invention includes a chip member having a pillar group composed of a plurality of pillars protruding from the surface of a recess, and a peripheral wall portion erected on the outer peripheral portion of the surface of the chip member, micro-objects in the sample liquid injected into the recess can be captured by the plurality of pillars. Therefore, the capture of micro-objects in the sample liquid can be easily realized with a simple structure.

Brief Description of the Drawings

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Embodiments for Carrying Out the Invention

[0017] Embodiment 1. Based on FIGS. 1 to 11, the micro-object capture system according to Embodiment 1 will be described. For convenience of explanation, in each figure, each component member includes an example different from the actual vertical, horizontal, and height ratios and sizes, and is shown in a simplified manner as appropriate. In each figure, different scales may be adopted for each component member from the viewpoint of visibility and the like. In each figure, for the purpose of avoiding complication of the drawing, a part of the reference numerals indicating each component member may be omitted. The same applies to each of the figures described later.

[0018] As illustrated in FIG. 1, the micro-object capture system 100 includes a micro-object capture device 110 and an injection device 120. The micro-object capture device 110 includes a chip member 10 and a peripheral wall portion 30. The chip member 10 has a concave portion 21 on the front side. The chip member 10 has a pillar group 22 composed of a plurality of pillars P protruding from the surface of the concave portion 21. Hereinafter, the concave portion 21 and the pillar group 22 formed therein are collectively referred to as a "capture portion 20". In FIGS. 1 to 5, the pillar group 22 is described as one block, but each pillar P constituting the pillar group 22 is a columnar member.

[0019] The concave portion 21 is formed inside the outer peripheral portion on the surface of the chip member 10. In the examples of FIGS. 1 to 5, the concave portion 21 is formed in a rectangular shape in plan view and is recessed by a predetermined height from the outer peripheral portion of the chip member 10. More specifically, the depth F of the concave portion 21 is equal to the height T of each pillar P. The pillar group 22 in the first embodiment includes four or more pillars P. The pillar group 22 captures the micro-objects S contained in the sample liquid injected into the concave portion 21 at the central portion between four adjacent pillars P. The micro-objects S include various cells, cell organelles, cell secretions, microorganisms, droplets, microgels, crystals, and the like. The dimensions of the chip member 10 are not particularly limited, but for example, it may be 5 mm to 100 mm in length (corresponding to the height M in FIGS. 3 to 5), 5 mm to 100 mm in width, and 0.05 mm to 10.00 mm in thickness. The height N of the peripheral wall portion 30 may be longer than the height T of the pillar P and can be arbitrarily set and changed as appropriate.

[0020] Here, the four pillars P adjacent to each other refer to, for example, four pillars P in a positional relationship like the region Z (FIG. 7) of FIG. 6. That is, among the four pillars P adjacent to each other, each pillar P is adjacent to the other three pillars P respectively, and a space is secured at the center of the four pillars P. In the pillar group 22 including a large number of pillars P as shown in FIG. 6, each pillar P is adjacent to eight pillars P respectively (excluding the pillars P arranged on the outermost side), and there are four spaces around for capturing the minute object S. FIG. 7 virtually shows the minute object S staying at the central portion (hereinafter also referred to as the capturing portion) of the four pillars P adjacent to each other.

[0021] The chip member 10 has a recovery portion 40 into which the sample liquid injected into the recess 21 flows. Further, the chip member 10 has a discharge flow path 15 connecting between the recess 21 and the recovery portion 40. The height of the discharge flow path 15 is equal to the depth F of the recess 21.

[0022] The peripheral wall portion 30 stands on the outer peripheral portion on the surface of the chip member 10 and surrounds the pillar group 22 from three directions. That is, the peripheral wall portion 30 is formed in a U-shaped (U-shaped with a right angle at the corner) in plan view. Note that the same shape as in the plan view appears in the cross section of the peripheral wall portion 30. The peripheral wall portion 30 in each figure is composed of a back wall 31 and a pair of side walls 32 connected to each end of the back wall 31.

[0023] Further, the minute object capturing device 110 has a partition portion 35 whose both side portions are sandwiched by two opposing side walls 32 of the peripheral wall portion 30 and surrounds the pillar group 22 together with the peripheral wall portion 30. The peripheral wall portion 30 of the first embodiment has notches 3k at positions facing each other on the pair of side walls 32. The notch 3k is a portion cut out from the upper end to the lower end of the side wall 32, and its cross-sectional shape is the same as the outer periphery of each side portion of the partition portion 35. The notch 3k enables the side portion of the partition portion 35 to be fitted. In each figure, the width of the notch 3k is substantially equal to the width of the partition portion 35.

[0024] That is, the partition portion 35 is fitted into the respective notches 3k of the pair of side walls 32 of the peripheral wall portion 30 at both sides, and together with the peripheral wall portion 30, surrounds the four sides of the pillar group 22. The partition portion 35 is aligned by the user so that both sides are at the upper part of each notch 3k, slid downward, and clamped by the two side walls 32.

[0025] The partition portion 35 in each figure has a plate-shaped base portion 35a and a plug portion 35b provided on one of the two end faces of the base portion 35a. Here, the two end faces of the base portion 35a are the faces facing upward or downward in a state where the partition portion 35 is clamped by the peripheral wall portion 30, respectively. The plug portion 35b is for opening and closing the discharge flow path 15, and the height C of the plug portion 35b is substantially equal to the height of the discharge flow path 15.

[0026] Figures 2 and 3 illustrate a state in which the partition portion 35 with the plug portion 35b facing upward is inserted into the locations of the pair of notches 3k of the peripheral wall portion 30. In the micro-particle capturing device 110 in this state, since the discharge flow path 15 is open, the sample liquid injected into the capturing portion 20 flows out to the recovery portion 40 through the discharge flow path 15 (see the white arrow). Therefore, even if the liquid level W of the sample liquid temporarily exceeds the height of the discharge flow path 15, if the injection of the sample liquid is interrupted, the liquid level W will become equal to the height of the discharge flow path 15 after a certain period of time.

[0027] Figure 4 is an explanatory diagram illustrating a state in which the partition portion 35 with the plug portion 35b facing downward is inserted into the locations of the pair of notches 3k of the peripheral wall portion 30. In the micro-particle capturing device 110 in this state, since the discharge flow path 15 is blocked, the sample liquid injected into the capturing portion 20 does not flow into the recovery portion 40 and stays in the region surrounded by the peripheral wall portion 30 and the partition portion 35 (hereinafter, also referred to as the capturing region). Therefore, the higher the sample liquid is injected into the capturing portion 20, the higher the liquid level W rises.

[0028] FIG. 5 is an explanatory diagram illustrating a state in which the partition portion 35 is removed from the peripheral wall portion 30. In the micro-object capturing device 110 in this state, since the side opposite to the back wall 31 in the peripheral wall portion 30 is open, all the sample liquids except for the micro-objects S captured at the upper part of the pillar group 22 and the micro-objects S caught between the side portions of the pillar group 22 flow into the recovery portion 40 (see the white arrows). The sample liquid accumulated in the recovery portion 40 may be pumped out using a pump or the like. The sample liquid pumped out from the recovery portion 40 may be injected into the capture portion 20 again at an arbitrary timing.

[0029] When the four sides of the capture portion 20 are completely closed as shown in FIG. 4, if evaporation is not considered, the amount of the sample liquid in the capture region can be maintained. Therefore, the usage amount of the sample liquid can be suppressed. If the upper ends of the peripheral wall portion 30 and the partition portion 35 are covered with a wrap or the like so as to cover the entire capture portion 20 to completely seal the capture region, evaporation of the sample liquid can be suppressed, and thus the amount of the sample liquid in the capture region can be maintained. The state as shown in FIG. 4 is useful when culturing cells as the micro-objects S captured by the pillar group 22.

[0030] However, among the micro-objects S, some have a large specific gravity and are likely to sink, while some have a small specific gravity and are likely to float. And in the state where the four sides of the capture portion 20 are completely closed as shown in FIG. 4, the micro-objects S that are likely to float float on the relatively upper part of the sample liquid and are difficult to drop to the height of the pillar group 22. Therefore, stable capture by the pillar group 22 is not easy. In this regard, in the state where the discharge channel 15 is open as shown in FIG. 3, since a flow toward the discharge channel 15 located below can be formed in the sample liquid in the capture region, the micro-objects S that are likely to float are induced by the flow and move downward. That is, in the state where the discharge channel 15 is open, even for the micro-objects S that are likely to float, capture by the pillar group 22 becomes easy.

[0031] The sample liquid may be directly injected from above the trapping section 20 or the like, but the micro-object trapping device 110 may be provided with an injection hole h1 for injecting the sample liquid in at least one of the chip member 10 and the peripheral wall section 30. FIGS. 1 and 2 show an example in which the injection hole h1 is provided in the chip member 10, and this injection hole h1 is formed across from the outer surface of the chip member 10 to the side surface of the recess 21. One end of a tubular injection port 1p is connected to the injection hole h1. The injection hole h1 is preferably provided at a location near the back wall 31 in the chip member 10 or the peripheral wall section 30.

[0032] When injecting the sample liquid from above the trapping section 20, since the sample liquid accumulated in the trapping region is likely to be wavy, it is not easy to stably maintain the state of capturing the micro-object S, such as the micro-object S that has been once captured moving. In this regard, if the sample liquid is injected into the trapping section 20 through the injection hole h1 from the injection port 1p, the control of the injection amount becomes easy, and the waviness of the liquid surface and the like can be suppressed, so that the micro-object S can be stably captured and stationary.

[0033] In the micro-object trapping system 100 of FIG. 1, an injection device 120 including a reservoir (not shown) for storing the sample liquid is connected to the other end of the injection port 1p. The injection device 120 of the first embodiment includes an electroosmotic flow pump 80 that sends the sample liquid in the reservoir to the recess 21 through the injection port 1p. Although not shown in any figure, the electroosmotic flow pump 80 has a main body portion formed of resin and a driving portion partially or entirely enclosed in the main body portion. The driving portion includes a pair of electrodes and a porous body sandwiched between the electrodes. The porous body is formed of, for example, porous ceramic.

[0034] When a voltage is applied across both ends of a porous body infiltrated with a liquid such as water or alcohols, a phenomenon (electroosmotic flow phenomenon) occurs in which the liquid in the porous body moves from one electrode side to the other electrode side, and the flow of the liquid generated by this phenomenon is called electroosmotic flow. The micro-object trapping system 100 is configured to indirectly extrude the sample liquid in the reservoir toward the micro-object trapping device 110 by electroosmotic flow.

[0035] The micro-object capturing device 110 may have a plurality of injection holes h1, and injection ports 1p may be attached to each of the plurality of injection holes h1. In this case, the plurality of injection holes h1 may be arranged in a well-balanced manner on either one or both of the chip member 10 and the peripheral wall portion 30. By doing so, the sample liquid can be circulated throughout the capture region at an early stage. For example, even when the discharge channel 15 is opened as shown in FIG. 3, it becomes easier to store the sample liquid throughout the capture region. Also, by suppressing the injection amount for each injection hole h1, the rippling of the sample liquid can be suppressed. The injection device 120 may be connected to the plurality of injection holes h1 via one injection port 1p. The micro-object capturing system 100 may have a plurality of injection devices 120 connected to one or more injection holes h1.

[0036] The pillar group 22 may be formed by arranging a plurality of pillars P having the same shape and size at equal intervals. In this case, the plurality of pillars P constituting the pillar group 22 may be arranged in a rectangular lattice pattern with a preset interval (predetermined interval). That is, in a plan view, the plurality of pillars P may be arranged such that the centers of each are located at the intersections of the vertical and horizontal lines of a lattice virtually arranged on the surface of the recess 21. In this way, when the plurality of pillars P are arranged in rows vertically and horizontally in a plan view, a lattice-like flow path is formed in the capture unit 20 by the plurality of pillars P constituting the pillar group 22.

[0037] In short, the capture unit 20 separates the micro-objects S in the sample liquid and captures the micro-objects S by the pillar group 22. The pillar group 22 has a plurality of pillars P having shapes such as those illustrated in FIGS. 6 to 11, and can capture and stably locate the micro-objects S at the central portion between four adjacent pillars P. That is, the pillar group 22 can align and stationary a plurality of micro-objects S.

[0038] (The first shape of the pillar P) FIG. 6 is a plan view showing an example of the pillar P of the first shape, and FIG. 7 is a plan view showing four adjacent pillars P arranged in the region Z of FIG. 6. The pillar P of the first shape has a shape in which, while being based on a rectangular shape in plan view, the portions corresponding to the four corners of the rectangular shape are cut out in an arc shape. The radius of curvature of the arc shape in the notch can be appropriately adjusted not limited to the examples in each figure, and the size of the notch can also be appropriately changed not limited to the examples in each figure. Note that the center O of the pillar P in plan view and the adjacent centers Q which are the center positions of the four adjacent pillars P are for illustrative convenience.

[0039] In the pillar group 22 of FIG. 6, the four adjacent pillars P of the first shape each have a side surface on the side of the center (adjacent center Q) of the four pillars P cut out in an arc shape in cross-sectional view. In other words, the pillar P of the first shape has a shape (taper shape) that tapers from the center or a predetermined length outside the center toward each of the four adjacent pillars P in plan view.

[0040] The plurality of pillars P illustrated in FIG. 6 are arranged in a square lattice at a predetermined interval. And each pillar P has a shape in which, while being based on a square shape in plan view, the portions corresponding to the four corners of the square shape are cut out in an arc shape. In FIG. 7, by showing the square shape K that is the basis of the pillar P with a broken line, it is made clear that the portions corresponding to the four corners of the square shape are recessed in an arc shape. In the first embodiment, the cross-sectional shape of the pillar P is the same from the upper end to the lower end, that is, the plan view and the cross-sectional view of the pillar P appear the same.

[0041] (Second shape of pillar P) FIG. 8 is a plan view showing an example of the pillar P of the second shape, and FIG. 9 is a plan view showing another example of the pillar P of the second shape. The pillar P of the second shape has a rectangular shape in plan view. The rectangular shape in plan view of the pillar P of the second shape includes shapes with chamfered corners (C chamfered) and shapes with rounded corners (R chamfered) as shown in FIG. 9.

[0042] The plurality of pillars P illustrated in FIG. 8 are arranged in a square lattice at a predetermined interval. The plurality of pillars P illustrated in FIG. 9 include a subgroup Ga and a subgroup Gb. The plurality of pillars P constituting the subgroup Ga and the subgroup Gb are each arranged in a square lattice at a predetermined interval. In the subgroup Ga and the subgroup Gb, the shapes and sizes of the constituent pillars P are equal, and the distance m between the pillars P is different. The distance m between the pillars P is the distance between the two closest adjacent pillars P.

[0043] More specifically, the vertical and horizontal lengths a1 of the pillar P of the subgroup Ga are equal to the vertical and horizontal lengths a2 of the pillar P of the subgroup Gb, and the distance m2 between the pillars P of the subgroup Gb is longer than the distance m1 between the pillars P of the subgroup Ga. From FIG. 9, it can be seen that the sizes of the capture locations are different between the subgroup Ga and the subgroup Gb. Thus, the pillar group 22 can capture minute objects S of different sizes for each region by providing a plurality of subgroups G with different distances m between the pillars P. For example, the length a1 and the length a2 are set to 80 μm, the distance m1 is set to 20 μm, and the distance m2 is set to 22 μm, but this is just an example.

[0044] (The third shape of the pillar P) FIG. 10 is a plan view showing an example of the pillar P of the third shape. The pillar P of the third shape is circular or elliptical in plan view. FIG. 10 illustrates a plurality of pillars P that are circular in plan view. The plurality of pillars P illustrated in FIG. 10 are arranged in a square lattice at a predetermined interval. The diameter a of the pillar P can be changed as appropriate.

[0045] The pillar group 22 can change the size of the object to be captured by a plurality of pillars P by changing the size (including the outer shape) of the capture location. The size of the capture location can be adjusted by changing the distance m between the pillars P. The distance m between the pillars P can be adjusted by changing the center-to-center distance X of the pillars P in a plan view, and can also be adjusted by changing the vertical and horizontal lengths a (diameter a) of the pillars P, or can be adjusted by combining these. The size of the capture location can also be adjusted by changing the shape of the pillars P.

[0046] 〔Regarding subgroups〕 The pillar group 22 may include four or more pillars P and may have a plurality of subgroups G in which the shapes and sizes of the pillars P are unified. The plurality of pillars P constituting the subgroup G may be arranged in a rectangular lattice at a predetermined interval, and it is more preferable to arrange them in a square lattice. The pillar group 22 may include four or more pillars P and may have a plurality of subgroups G in which the distance m between the pillars P is unified. The plurality of pillars P constituting the subgroup G are preferably arranged in a rectangular lattice at a predetermined interval, and it is more preferable to arrange them in a square lattice. In the pillar group 22, subgroups G having different shapes and sizes of the pillars P may be mixed, and subgroups G having different distances m between the pillars P may be mixed. The number of pillars P constituting the subgroup G can be arbitrarily set and changed as appropriate.

[0047] Based on the injection hole h1, the distance m between the pillars P in the upstream subgroup G of the pillar group 22 is longIt may be configured as follows. For example, the pillar group 22 may be formed such that each pillar P in the downstream sub-group G is thinner than each pillar P in the upstream sub-group G with respect to the injection hole h1. That is, the pillar group 22 may be configured such that the size of the pillar P in plan view is smaller in the downstream sub-group G than in the upstream sub-group G with respect to the injection hole h1. In this way, the micro-object capturing device 110 can be adjusted to capture relatively large micro-objects S in the upstream sub-group G and relatively small micro-objects S in the downstream sub-group G. Such a configuration can be utilized for state management for each size of the micro-object S and the like.

[0048] 〔Device for identifying regions〕 The pillar group 22 may be configured to distinguish at least one pillar P from other pillars P so that it is easier for the user to identify a specific region. Such a configuration is useful when performing some processing on the captured micro-object S and observing the subsequent progress.

[0049] FIG. 11 is a plan view illustrating a pillar group 22 including a pillar P with an identifier D attached thereto. In the example of FIG. 11, the identifier D is attached to the surfaces of four pillars P. The identifier D 1 is associated with a management region R 1 and the identifier D 2 is associated with a management region R 2 and the identifier D 3 is associated with a management region R 3 and the identifier D 4 is associated with a management region R 4 and the identifier D 1 ~R 4 is a region including four or more pillars P and is set for managing the captured micro-objects S for each region.

[0050] The pillar group 22 may be entirely divided into a plurality of management areas, a part of the areas may be divided into a plurality of management areas, or only one management area may be set. In the example of FIG. 11, an identifier D unique to the management area is attached to the pillar P arranged at the corner of each management area. The identifier D is not limited to the mode illustrated in FIG. 11. That is, the identifier D can be constituted by characters, symbols, figures, colors, or a combination of some of these.

[0051] 〔Manufacturing process example of micro-object capturing device〕 The micro-object capturing device 110 can be manufactured, for example, by the following steps. <Creation of a mold for the chip member 10> (1) Prepare a design drawing. (2) Produce a photomask by laser drawing. (3) Apply a resist to the silicon wafer using a spin coater or the like. (4) Perform UV irradiation using an aligner or the like. (5) Perform etching (completion of the mold). <Creation of the chip member 10> (6) Apply liquid silicon to the mold. (7) Heat in an oven (for example, at 120 °C for 1 hour) (8) Release the solidified silicone rubber from the mold. (9) Partially cut the released silicone rubber and drill the necessary holes (completion of the chip member 10) <Creation of the peripheral wall portion 30 and the partition portion 35> (10) Produce the peripheral wall portion 30 and the partition portion 35 by cutting from a silicone block. <Bonding of the chip member 10 and the peripheral wall portion 30> (11) Bond the chip member 10 and the peripheral wall portion 30 together by plasma treatment or the like. (12) Perform a hydrophilic treatment on the entire surface of the chip member 10 or on the entire surface from the capturing portion 20 including the pillar group 22 to the recovery portion 40.

[0052] In addition, the steps (1) to (12) may be executed by appropriately swapping them. The step (12) may be omitted. However, when an aqueous sample solution such as cells is injected into the capture part 20 of the chip member 10, water may be repelled on the surface of the chip member 10, and the sample solution may not flow well. In this regard, if hydrophilicity is imparted to at least the surface of the chip member 10, the aqueous sample solution can flow smoothly to every corner of the capture part 20, and the minute substances S in the sample solution can be efficiently captured by the pillar group 22. A hydrophilic treatment may be applied to the inner surface of the peripheral wall part 30.

[0053] The chip member 10 is preferably made of a material having a polarity close to that of the sample solution to be used. The chip member 10 may be a smooth plate. If the chip member 10 has transparency and low fluorescence properties, observation with a microscope can be suitably performed. The material of the chip member 10 is not particularly limited. For example, transparent plastics such as PDMS (cycloolefin copolymer), PS (polystyrene), PMMA (polymethyl methacrylate: acrylic resin), PP (polypropylene), polyolefin, PC (polycarbonate), PE (polyethylene), or glass or quartz can be suitably used. As the materials of the peripheral wall part 30 and the partition part 35, transparent plastics such as PDMS, PS, PMMA, PP, polyolefin, PC, PE, or glass or quartz can be suitably used. When observing the pillar group 22 with a microscope, at least the recess 21 is preferably made transparent, and the pillar group 22 may also be made transparent.

[0054] As described above, the micro-object capture device 110 in the first embodiment includes a chip member 10 having a pillar group 22 composed of a plurality of pillars P protruding from the surface of the recess 21 formed on the front side, and a peripheral wall portion 30 erected on the outer peripheral portion of the surface of the chip member 10 and surrounding the pillar group 22 from three directions. Further, the micro-object capture device 110 has a partition portion 35 that sandwiches both side portions by a pair of opposing side walls 32 of the peripheral wall portion 30 and surrounds the four sides of the pillar group 22 together with the peripheral wall portion 30. And the upper end of the peripheral wall portion 30 and the upper end of the partition portion 35 are higher than the positions of the upper ends of the respective pillars P. Therefore, the micro-objects S in the sample liquid injected into the recess 21 can be efficiently captured by the plurality of pillars P.

[0055] The number of pillars P constituting the pillar group 22 may be two or three. Even with such a configuration, the pillar group 22 can capture the micro-objects S by the inner surface of the recess 21 and the two or three pillars P. If there are three pillars P, by arranging them at the vertices of a triangle, the micro-objects S can be captured even at the central portion thereof. However, it is more preferable that the pillar group 22 includes four or more pillars P. The pillar group 22 including four or more pillars P captures the micro-objects S contained in the sample liquid injected into the recess 21 at the central portion between four mutually adjacent pillars P.

[0056] The plurality of pillars P constituting the pillar group 22 may be arranged randomly, but it is preferable to arrange them in a rectangular lattice pattern at a preset interval. In this way, since it is possible to align and then stationary the micro-objects S above the pillar group 22, the management of the micro-objects S becomes easy. It is more preferable that the plurality of pillars P constituting the pillar group 22 are arranged in a square lattice pattern at a preset interval.

[0057] The pillar group 22 may have a plurality of sub - groups G in which the shapes and sizes of the pillars P are unified. The plurality of pillars P constituting the sub - group G are preferably arranged in a rectangular lattice pattern at a preset interval, and more preferably in a square lattice pattern. In the pillar group 22, sub - groups G with different pillar sizes may be mixed. In the pillar group 22, sub - groups G with different pillar shapes may be mixed. The pillar group 22 may have a plurality of sub - groups G in which the distance m between the pillars P is unified. The plurality of pillars P constituting the sub - group G are preferably arranged in a rectangular lattice pattern at a preset interval, and more preferably in a square lattice pattern. In the pillar group 22, sub - groups G with different distances m between the pillars P may be mixed. Thus, by mixing sub - groups G with different configurations in the pillar group 22, it becomes possible to adjust the minute object S to be captured for each sub - group G.

[0058] At least one of each pillar P constituting the pillar group 22 may have an identifier on its surface. With such a configuration, it becomes easy to manage the minute object S captured around the identifier. At least one of the chip member 10 and the peripheral wall portion 30 may be provided with an injection hole h1 for injecting the sample liquid. If the sample liquid is injected from the injection hole h1 into the recess 21, waves are less likely to occur in the sample liquid within the capture region, so that the capture and stationary placement of the minute object S by the pillar group 22 can be performed stably and accurately. If the sample liquid is injected from the injection hole h1 using the injection device 120 including the electroosmotic flow pump 80, a small amount of the sample liquid can be injected into the capture portion 20 drop by drop, so that the liquid volume can be accurately controlled and a more stable stationary placement of the minute object S can be realized.

[0059] Of the plurality of pillars P that make up the pillar group 22, at least four of them may have a shape that is mainly rectangular in plan view, with the portions corresponding to the four corners of the rectangle being cut out in an arc shape. Of the plurality of pillars P that make up the pillar group 22, at least four of them may have a shape that is mainly square in plan view, with the portions corresponding to the four corners of the square being cut out in an arc shape. In this way, a shape along the surface of the spherical microparticle S appears at the central portion of four adjacent pillars P, making it easier to capture the microparticle S and enabling it to be stably placed. That is, a cylindrical void (a void into which a cylinder fits snugly) surrounded by the side surfaces of each pillar P on the adjacent center Q side is formed at the center of four adjacent pillars P, facilitating the catching of the microparticle S.

[0060] With respect to the pillar group 22, based on the injection hole h1, the distance m between the pillars P in the upstream sub-group G is greater than the distance m between the pillars P in the downstream sub-group G. long It may be made like this. In this way, it is possible to adjust the size of the microparticle S to be captured for each region, such as capturing a relatively large microparticle S in the upstream sub-group G and a relatively small microparticle S in the downstream sub-group G. For example, for a pillar group 22 including a plurality of sub-groups G in which a plurality of pillars P are arranged in a rectangular grid, based on the injection hole h1, the size of each pillar P in the downstream sub-group G may be smaller than the size of each pillar P in the upstream sub-group G. In this case, based on the injection hole h1, the center-to-center distance X of the pillars P in the downstream sub-group G may be shorter than the center-to-center distance X of the pillars P in the upstream sub-group G. For a pillar group 22 including a plurality of sub-groups G in which a plurality of pillars P of the first shape are arranged in a rectangular grid, without changing the size of the pillar P, the radius of curvature of the arc shape in the corner notch may be made smaller in the downstream sub-group G than in the upstream sub-group G.

[0061] The pair of side walls 32 may have notches 3k on both sides of the partition portion 35 that are fitted into positions facing each other. If the partition portion 35 is fitted into the pair of notches 3k, the partition portion 35 can be stably fixed. However, the notches 3k may not be provided on the pair of side walls 32, and the lateral width of the partition portion 35 may be made equal to the distance between the pair of side walls 32, and the partition portion 35 may be simply configured to be clamped by the pair of side walls 32.

[0062] The micro-object capture device 110 may be provided with a recovery portion 40 into which the sample liquid injected into the recess 21 flows. In this way, it is not necessary to recover the sample liquid from the capture region each time, and the injection of the sample liquid into the capture region can be continuously performed. The partition portion 35 may be composed of a base portion 35a and a plug portion 35b. In this way, when capturing the floating micro-object S on the pillar group 22, it can be used in a state where the discharge channel 15 is open (see Fig. 3), and when it is desired to observe the micro-object S captured on the pillar group 22 for a long time, it can be used in a state where the discharge channel 15 is blocked (see Fig. 4), etc., enabling proper use according to the situation. In Figs. 1 to 5, the chip member 10 having the discharge channel 15 connecting the recess 21 and the recovery portion 40 is illustrated, but the chip member 10 may be configured to directly connect the recess 21 and the recovery portion 40 without providing the discharge channel 15.

[0063] The chip member 10 may be one in which a hydrophilic treatment is applied to the surface including the pillar group 22 surrounded by the peripheral wall portion 30. With such a configuration, the aqueous sample liquid can flow smoothly to every corner of the capture portion 20, so that the micro-object S in the sample liquid can be efficiently and accurately captured on the pillar group 22. Also, the sample liquid can flow smoothly from the capture portion 20 toward the recovery portion 40.

[0064] By the way, these days, it has become possible to pick only one target while observing minute object S under a microscope. In the field where such picking is performed, there is a demand to open the upper part of the container housing the minute object S so that a minute glass needle can access it. In this regard, in the minute object capturing device 110 of the first embodiment, since the upper part of the capturing region surrounded by the peripheral wall portion 30 and the partition portion 35 is open, the user can easily pick the minute object S captured by the pillar group 22.

[0065] Also, when assuming cell culture in a minute space on a microscope, it becomes a problem that the liquid evaporates immediately. In addition, in order to pick the target minute object S from the container, it is necessary to open the upper part of the container, but also at that time, there is a problem that the culture medium and the droplet oil evaporate and dry immediately. Furthermore, when trying to pick only one target under microscope observation from the cells on the plate-shaped chip, since it is necessary to handle it within the limited space of the microscope stage, it is necessary to miniaturize the chip itself. Therefore, the solution containing cells or the oil containing the w / o droplet has a limited capacity on the chip. In particular, the oil for droplets has high volatility and evaporates immediately. When the upper surface of the chip dries, picking becomes difficult and it may lead to breakage of the droplet. In this regard, the minute object capturing device 110 of the first embodiment can block the capturing region by covering the upper parts of the peripheral wall portion 30 and the partition portion 35. Therefore, evaporation of the sample liquid can be suppressed, and the user can perform stable culture and its observation.

[0066] <Modification Example A> Based on FIGS. 12 and 13, a minute object capturing system according to Modification Example A of the first embodiment will be described. In each figure, for convenience of explanation, each constituent member is appropriately simplified, and the ratio and size of the vertical, horizontal, and height of each constituent member are adjusted and shown. For the minute object capturing system of this Modification Example A, the constituent members equivalent to those of the minute object capturing system 100 in the main text described above are denoted by the same reference numerals, and the overlapping explanations are omitted or simplified.

[0067] The micro-object capture system 100 of this Modification Example A includes a micro-object capture device 110A and an injection device 120. The micro-object capture device 110A has a group partition portion 38 that is sandwiched on both sides by a pair of side walls 32 of the peripheral wall portion 30 and divides the capture region. The group partition portion 38 in each figure divides the pillar group 22 into two regions.

[0068] FIG. 13 illustrates a micro-object capture device 110A with the partition portion 35 and the group partition portion 38 removed. The peripheral wall portion 30 has notches 4k at positions facing each other on the pair of side walls 32. The notch 4k is a portion cut out from the upper end to the lower end of the side wall 32, and its cross-sectional shape is the same as the outer periphery of each side portion of the partition portion 35. The notch 4k enables the side portion of the group partition portion 38 to be fitted. In each figure, the width of the notch 4k is substantially equal to the width of the group partition portion 38. The notch 4k is provided at a position facing the pillar group 22 on the pair of side walls 32. In the example of each figure, a pair of notches 4k are provided around approximately the center of the pillar group 22. The group partition portion 38 is aligned by the user so that both side portions come above each notch 4k, slid downward, and sandwiched by the two side walls 32. The micro-object capture device 110A has a pair of partition bases 4s that support the group partition portion 38 inserted between the pair of notches 4k. The micro-object capture device 110A is manufactured in the same manner as the micro-object capture device 110.

[0069] As described above, the micro-object capture device 110A of this Modification Example A has a group partition portion 38 that is sandwiched on both sides by a pair of opposing side walls 32 of the peripheral wall portion 30 above the pillar group 22. Each region divided by the group partition portion 38 is connected by the gap between each pillar P between the lower end of the group partition portion 38 and the bottom of the concave portion 21. However, the micro-objects S captured above the pillar group 22 basically cannot move back and forth between the regions, so the micro-objects S can be managed for each region. The pillar group 22 may change the distance m between the pillars P for each region divided by the group partition portion 38. If the distance m between the pillars P in the upstream region is made longer than the distance m between the pillars P in the downstream region, it becomes possible to capture the micro-objects S that were not captured in the upstream region in the downstream region.

[0070] A pair of notches 4k may be provided at a plurality of locations on the peripheral wall portion 30. In the case of such a configuration, the pillar group 22 may be divided into two regions using one group partition portion 38, or the pillar group 22 may be divided into three or more regions using a plurality of group partition portions 38. The notches 4k may not be provided on the pair of side walls 32, and the lateral width of the group partition portion 38 may be made equal to the distance between the pair of side walls 32, that is, the pair of partition bases 4s may be eliminated, and the group partition portion 38 may be configured to be sandwiched by the pair of side walls 32 at an arbitrary position in the capture region. Also in the case of such a configuration, the minute object capture device 110A may be configured to divide the capture region into three or more regions by a plurality of group partition portions 38.

[0071] Embodiment 2. Based on FIGS. 14 to 18, the minute object capture system according to Embodiment 2 will be described. In each figure, for convenience of explanation, each component is appropriately simplified, and the ratio and size of the vertical, horizontal, and height of each component are adjusted and shown. In each figure, a part of the reference numerals indicating each component may be omitted for the purpose of avoiding complication of the drawing.

[0072] The minute object capture system 100 of the present Embodiment 2 includes an injection device 120 together with the minute object capture device 210, but the illustration of the injection device 120 is omitted. For the minute object capture device 210, the same reference numerals are given to the components equivalent to those of the minute object capture device 110 of Embodiment 1, and the overlapping explanations are omitted or simplified.

[0073] The chip member 10 has a capture part 20, a partition table 16, a filter part 50, and a recovery part 40. The filter part 50 is arranged downstream of the pillar group 22 and includes a plurality of columnar parts L. More specifically, the chip member 10 has a downstream recess 51 on the front side and a column group 52 composed of a plurality of columnar parts L protruding from the surface of the downstream recess 51. That is, the filter part 50 is composed of the downstream recess 51 and the column group 52 formed therein. In the filter part 50, the distance between the columnar parts L in the column group 52 is shorter than the distance m between the pillars P in the pillar group 22. Therefore, a part of the minute objects S that have passed through the capture part 20 is caught by the column group 52, and smaller minute objects S flow into the recovery part 40.

[0074] The recovery part 40 is arranged downstream of the filter part 50 and is the part where the sample liquid injected into the recess 21 and passing through the filter part 50 flows in through the discharge channel 15. The recovery part 40 in each figure has its bottom closed by the lower lid 41 and can store liquid. However, the recovery part 40 may be a groove-like one provided in the chip member 10 as in FIG. 1. The partition table 16 is the part that partitions between the recess 21 and the downstream recess 51, and as shown in FIG. 16, the end face of the partition part 35 held by the peripheral wall part 30 abuts.

[0075] In the peripheral wall part 330, an adjustment hole h2 for adjusting the amount of the sample liquid injected into the recess 21 is provided at a position higher than the upper end of each pillar P in the pillar group 22. If there is the adjustment hole h2, when the liquid level of the sample liquid accumulated in the capture area reaches the height of the adjustment hole h2, the sample liquid flows out to the outside from the adjustment hole h2, so that the amount of the sample liquid staying in the capture area can be limited. A tubular adjustment port 2p may be connected to the adjustment hole h2. The adjustment port 2p may be connected to a container for storing the sample liquid flowing out from the adjustment hole h2 or may be connected to the reservoir of the injection device. A suction device (not shown) including a pump may be connected to the adjustment port 2p to suck the sample liquid in the capture area from the outside.

[0076] FIG. 16 illustrates a state in which the partition portion 35 is inserted into the locations of the pair of notches 3k in the peripheral wall portion 30. In this way, when the partition portion 35 is closed, the sample liquid injected into the capture portion 20 stays in the capture region. If a lid is placed on the capture region in this state, evaporation of the sample liquid can be prevented and the liquid volume can be maintained. FIG. 17 illustrates a state in which the partition portion 35 is removed from the peripheral wall portion 30. In this state, the sample liquid accumulates in the capture portion 20 up to the height of the partition base 16. After closing the partition portion 35 as in FIG. 16 to retain the sample liquid in the capture region and then removing the partition portion 35 as in FIG. 17, the sample liquid passes through the partition base 16 and the filter portion 50 and flows into the recovery portion 40, and the sample liquid at the same height as the partition base 16 remains in the capture portion 20.

[0077] FIG. 18 is a schematic diagram illustrating how minute objects S in a sample liquid are separated by size together with the pillar group 22, the column group 52, and the recovery portion 40 in the minute object capture device 210. FIG. 18 illustrates a state in which relatively large minute objects S in the sample liquid injected into the capture portion 20 are captured by the pillar group 22 and minute objects S smaller than the captured ones flow out of the capture portion 20. In FIG. 18, among the minute objects S in the sample liquid that has flowed out of the capture portion 20, minute objects S having a diameter larger than the gap between the columnar portions L stay in the filter portion 50, and minute objects S having a diameter smaller than the gap between the columnar portions L flow from the filter portion 50 into the recovery portion 40.

[0078] In FIG. 18, a triangular prism-shaped columnar portion L is illustrated, but it is not limited thereto. The shape of the columnar portion L may be other prismatic shapes such as a quadrangular prism, a pentagonal prism, a hexagonal prism, etc., may be a cylindrical shape, may be an elliptical prism shape, etc., and the column group 52 may be a combination of columnar portions L having different shapes. Other configurations of the minute object capture device 210 are the same as those of the minute object capture devices 110 and 110A in the first embodiment. The minute object capture device 210 is manufactured in the same manner as the minute object capture device 110.

[0079] As described above, the micro-object capturing device 210 has the chip member 10, the peripheral wall portion 30, and the partition portion 35, and the upper ends of the peripheral wall portion 30 and the partition portion 35 are higher than the upper ends of the respective pillars P. Therefore, the micro-objects S in the sample liquid injected into the recess 21 can be efficiently captured by the plurality of pillars P.

[0080] Further, the micro-object capturing device 210 of the second embodiment is disposed downstream of the pillar group 22 and has a filter portion 50 including a plurality of columnar portions L. In the filter portion 50, the distance between the columnar portions L is shorter than the distance m between the pillars P in the pillar group 22. That is, the micro-object capturing device 210 is configured to separate the micro-objects S captured by the pillar group 22 from other micro-objects S in the capturing portion 20, and further separate the micro-objects S caught between the columnar portions L from other micro-objects S in the filter portion 50. Therefore, the sample liquid containing relatively small micro-objects S that have passed between the columnar portions L is stored in the recovery portion 40. The sample liquid flowing into the recovery portion 40 can be provided to researchers, businesses, etc. who desire the small micro-objects S.

[0081] The peripheral wall portion 30 may be provided with an adjustment hole h2 for adjusting the amount of the sample liquid injected into the recess 21 at a position higher than the upper end of each pillar P in the pillar group 22. Thereby, the amount of the sample liquid staying in the capture region can be limited, and it is possible to prevent the sample liquid from overflowing outside the peripheral wall portion 30. The micro-object capturing device 210 may have a plurality of adjustment holes h2. The micro-object capturing device 210 can also achieve the same effects as the micro-object capturing devices 110 and 110A of the first embodiment described above. The configuration related to the group partition portion 38 of the modification example A can also be adopted for the micro-object capturing device 210. The adjustment hole h2 may be provided in the peripheral wall portion 30 of the micro-object capturing devices 110 and 110A of the first embodiment.

[0082] <Modification Example B> Based on FIGS. 19 to 23, a micro-object capturing system according to a modification example B of the second embodiment will be described. In each figure, for convenience of explanation, each component is appropriately simplified, and the ratio and size of the vertical, horizontal, and height of each component are adjusted and shown.

[0083] The microparticle capture system 100 of this modification example B has an injection device 120 together with the microparticle capture device 210A, but the illustration of the injection device 120 is omitted. For the microparticle capture device 210A, the same reference numerals are given to the constituent members equivalent to those of the microparticle capture devices 110 and 210, and redundant descriptions are omitted or simplified.

[0084] The microparticle capture device 210A has a pair of partition tables 17 that support a partition portion 35 inserted between a pair of notches 3k. Further, the microparticle capture device 210A has a communication flow path 18 that connects the concave portion 21 and the downstream concave portion 51. The communication flow path 18 is formed between a pair of partition tables 17. The partition portion 35 in this modification example B has a plate-shaped base portion 35a and a plug portion 35b provided on one of the two end faces of the base portion 35a. The plug portion 35b is for opening and closing the communication flow path 18, and the height C of the plug portion 35b is substantially equal to the height of the communication flow path 18. The shapes of the pair of partition tables 17 and the plug portion 35b are not limited to the examples in each figure, and various shapes can be adopted as long as the communication flow path 18 can be blocked by the plug portion 35b.

[0085] FIG. 21 illustrates a state in which the partition portion 35 with the plug portion 35b facing upward is inserted at the locations of the pair of notches 3k of the peripheral wall portion 30. In the microparticle capture device 210A in this state, since the communication flow path 18 is open, the sample liquid injected into the capture portion 20 passes through the filter portion 50 via the communication flow path 18 and flows into the recovery portion 40. As shown in FIG. 21, in the state where the communication flow path 18 is opened while closing the capture region with the partition portion 35, a downward flow toward the communication flow path 18 can be formed in the sample liquid in the capture region, so that it becomes easier to capture the floating microparticles S with the pillar group 22.

[0086] FIG. 22 illustrates a state in which a partition portion 35 with a plug portion 35b facing downward is inserted at the locations of a pair of notches 3k in the peripheral wall portion 30. In the micro-object capturing device 210A in this state, since the communication flow path 18 is blocked, the sample liquid injected into the capturing portion 20 stays in the capturing region. In the state where the communication flow path 18 is blocked as shown in FIG. 22, since there is no outflow of the sample liquid, this state is useful when it is desired to manage the micro-objects S captured by the pillar group 22 in the medium to long term. If a lid is placed over the capturing region, evaporation of the sample liquid can be prevented, enabling even more stable management of the micro-objects S.

[0087] FIG. 23 is an explanatory diagram illustrating a state in which the partition portion 35 is removed from the peripheral wall portion 30. In the micro-object capturing device 210A in this state, the side opposite to the back wall 31 in the peripheral wall portion 30 is open. Therefore, a part of the micro-objects S contained in the sample liquid injected into the capturing portion 20 is captured by the pillar group 22, a part of the micro-objects S that have passed through the pillar group 22 is caught by the filter portion 50, and the micro-objects S that have passed through the filter portion 50 flow into the recovery portion 40.

[0088] As described above, the chip member 10 has a communication flow path 18 that connects between the concave portion 21 and the filter portion 50. And the partition portion 35 has a plate-shaped base portion 35a and a plug portion 35b for opening and closing the communication flow path 18. Therefore, the micro-object capturing device 210A can be used selectively between the state as shown in FIG. 21 and the state as shown in FIG. 22 according to the properties of the micro-objects S and the treatment / management policy for the micro-objects S. The configuration related to the group partition portion 38 of Modification A can also be adopted in the micro-object capturing device 210A.

[0089] Embodiment 3. Based on FIG. 24, a micro-object capturing system according to Embodiment 3 will be described. In each figure, for the sake of convenience of explanation, each component member is appropriately simplified, and the ratio and size of the vertical, horizontal, and height of each component member are adjusted and shown. For the micro-object capturing system of this Embodiment 3, component members equivalent to those of the micro-object capturing system 100 of each of the above-described embodiments are denoted by the same reference numerals, and duplicate explanations are omitted or simplified.

[0090] The micro-object capture system 100 according to Embodiment 3 includes a micro-object capture device 310 and an injection device 120. The micro-object capture device 310, together with the chip member 10, has a peripheral wall portion 330 that stands on the outer peripheral portion of the surface of the chip member 10 and surrounds the pillar group 22 from four sides. The peripheral wall portion 330 in FIG. 24 is formed in a rectangular frame shape (a U shape) in plan view. That is, the peripheral wall portion 330 has a plate-shaped downstream wall 33 that faces the back wall 31, together with the back wall 31 and a pair of side walls 32.

[0091] The sample liquid may be pumped out from the region surrounded by the peripheral wall portion 330 (hereinafter also referred to as the capture region) by a pump or the like. However, as shown in FIG. 24, the micro-object capture device 310 may be provided with a discharge hole h3 in the chip member 10 for discharging the sample liquid from within the capture region. The discharge hole h3 illustrated in FIG. 24 is formed from the outer surface of the chip member 10 across the bottom surface of the recess 21. One end of a tubular discharge port 3p is connected to the discharge hole h3. The discharge hole h3 is preferably provided at a position away from the injection hole h1. Other configurations of the micro-object capture device 310 are the same as those of the micro-object capture device 110. The configuration related to the group partition portion 38 of Modification A can also be adopted for the micro-object capture device 310. The micro-object capture device 310 is manufactured in the same manner as the micro-object capture device 110.

[0092] As described above, the micro-object capture device 310 includes a chip member 10 provided with a pillar group 22 composed of a plurality of pillars P protruding from the surface of the recess 21 formed on the front side, and a peripheral wall portion 330 that stands on the outer peripheral portion of the surface of the chip member 10 and surrounds the pillar group 22 from four sides. And the upper end of the peripheral wall portion 330 is higher than the position of the upper end of each pillar P. Therefore, the micro-objects S in the sample liquid injected into the recess 21 can be efficiently captured by the plurality of pillars P.

[0093] The micro-object capturing device 310 may be configured without providing the discharge hole h3. However, if the discharge hole h3 is provided, the operation of discharging the sample liquid from the capturing space can be reduced. Further, when the sample liquid is being discharged from the discharge hole h3, a downward flow toward the discharge hole h3 can be formed in the sample liquid within the capturing region, making it easier to capture the floating micro-objects S with the pillar group 22. The discharge hole h3 or the discharge port 3p connected to the discharge hole h3 may be configured to be openable and closable. That is, the micro-object capturing system 100 may be provided with an opening / closing mechanism at the discharge hole h3 or the discharge port 3p. In this way, it becomes possible to properly use the state where the sample liquid within the capturing region flows out to the outside and the state where it does not flow out. A suction device (not shown) including a pump may be connected to the discharge port 3p to suck the sample liquid within the capturing region from the outside. If the suction force of the suction device is configured to be controllable, the discharge amount of the sample liquid can be controlled, and the flow of the sample liquid within the capturing region can also be controlled. The configuration related to the discharge hole h3 in the third embodiment can also be adopted in the micro-object capturing devices 110, 110A of the first embodiment and the micro-object capturing devices 210, 210A of the second embodiment.

[0094] 〔Example〕 Figs. 25 to 28 are examples showing a state where a sample liquid is dripped onto the capturing portion 20 of an actually manufactured micro-object capturing device and micro-objects are captured by the pillar group 22. Figs. 25 to 28 partially show the pillar group 22 in a state where the sample liquid is accumulated.

[0095] In the region shown in Fig. 25, sub-groups G 1 and sub-group G 2 and sub-group G 3 and sub-group G 4 appear. Sub-groups G 1 ~G 4 are each composed of pillars P 1 ~P 4 . In the pillar group 22, the sizes of the pillars P 1 , pillar P 2 , pillar P 3 , and pillar P 4 decrease in this order. The distance m between the pillars P is sub-group G 1, subgroup G 2 , subgroup G 3 , subgroup G 4 are getting shorter in this order.

[0096] In the region shown in FIG. 26, subgroup G 2 and subgroup G 3 and subgroup G 4 and subgroup G 5 and subgroup G 6 appear. Subgroups G 2 to G 5 are each composed of pillars P of the first shape 2 to P 5 . In pillar group 22, the sizes of pillars P 6 are getting smaller in the order of pillar P 6 to P 2 , pillar P 3 , pillar P 4 , pillar P 5 . The distance m between the pillars is getting shorter in the order of subgroup G 2 , subgroup G 3 , subgroup G 4 , subgroup G 5 .

[0097] In the region shown in FIG. 27, subgroup G 7 and subgroup G 8 and subgroup G 9 and subgroup G 10 and subgroup G 11 appear. Subgroups G 7 to G 10 are each composed of pillars P of the second shape 7 to P 10 . Subgroup G 11 is composed of a pillar P of the third shape 11 . In pillar group 22, the sizes of pillars P 7 , pillar P 8 , pillar P 9 , pillar P 10 are getting larger in this order. The distance m between the pillars is for subgroup G 7 , subgroup G 8 , subgroup G 9 , subgroup G10 They are getting longer in the order of

[0098] In the region shown in FIG. 28, there are subgroups G 12 and subgroup G 13 and subgroup G 14 and subgroup G 15 and subgroup G 16 appearing. Subgroups G 12 to G 16 are each composed of pillars P 12 to P 16 of the third shape. In the pillar group 22, the sizes of the pillars P 12 the pillar P 13 the pillar P 14 the pillar P 15 the pillar P 16 are getting larger in this order. The distance m between the pillars P is getting longer in the order of subgroup G 12 subgroup G 13 subgroup G 14 subgroup G 15 subgroup G 16

[0099] From FIGS. 25 to 28, it can be confirmed that a plurality of minute objects S are captured by the pillar group 22 in a state of being aligned vertically and horizontally. By using the minute object capturing device of each embodiment, just by dropping the sample liquid onto the capturing region, the minute objects S can be placed on the pillar group 22 to the extent shown in each figure at least without any particular adjustment. The minute object capturing device of each embodiment can be used in life science research, and can preferably align, stationary, and separate the minute objects S.

[0100] ​Each of the above-described embodiments is merely an example in the microparticle capturing device and the microparticle capturing system, and the technical scope of the present invention is not limited to these aspects. For example, the microparticle capturing device does not necessarily have to be rectangular in plan view, and may be circular or elliptical in plan view. However, the microparticle capturing device may have R chamfers at the corners. In any case, the microparticle capturing devices 110, 110A, 210, 210A of Embodiments 1 and 2 may be configured such that the capturing portion 20 is surrounded from all four sides by the peripheral wall portion 30 and the partition portion 35, and the microparticle capturing device 310 of Embodiment 3 may be configured such that the capturing portion 200 is surrounded from all four sides by the peripheral wall portion 330. The shape of the recovery portion 40 is not limited to the examples in each figure, and various shapes can be adopted. The shapes of the plug portion 35b, the discharge flow path 15, and the communication flow path 18 are not limited to the examples in each figure, and various shapes can be adopted. The partition portion 35 of Embodiment 1 and Modification Example B may be composed only of the base portion 35a without providing the plug portion 35b. The arrangements of the injection hole h1, the adjustment hole h2, and the discharge hole h3 in each figure are merely illustrative and can be changed as appropriate. Note that matters regarding the shape of the pillar P described based on FIGS. 6 to 11 are common to all embodiments.

Explanation of Reference Numerals

[0101] 1p injection port, 2p adjustment port, 3k notch, 3p discharge port, 4k notch, 4s, 16, 17 partition base, 10 chip member, 15 discharge flow path, 18 communication flow path, 20 capturing portion, 21 recess, 22 pillar group, 30, 330 peripheral wall portion, 31 back wall, 32 side wall, 33 downstream wall, 35 partition portion, 35a base portion, 35b plug portion, 38 group partition portion, 40 recovery portion, 41 lower lid, 45 discharge flow path, 50 filter portion, 51 downstream recess, 52 pillar group, 80 electroosmotic flow pump, 100 microparticle capturing system, 110 microparticle capturing device, 110A microparticle capturing device, 120 injection device, 200 capturing portion, 210 microparticle capturing device, 210A microparticle capturing device, 310 microparticle capturing device, 330 peripheral wall portion, C height, D, D 1 ~D 4 identifier, F depth, G, G 1 ~G 16, Ga, Gb subgroups, L columnar part, M, N, T height, O center, P, P 1 ~P 16 Pillar, Q adjacent center, R 1 ~R 4 Management area, S micro-object, W liquid level, X center-to-center distance of pillars, Z area, a longitudinal and transverse length (diameter) of pillar P, h1 injection hole, h2 adjustment hole, h3 discharge hole, m, m1, m2 distances.

Claims

1. a chip member including a pillar group consisting of a plurality of pillars protruding from a surface of a recess formed on a front side; a peripheral wall portion that is erected on an outer periphery of a surface of the tip member and surrounds the pillar group from three directions; a partition portion having both side portions sandwiched between a pair of opposing side walls of the peripheral wall portion and surrounding all four sides of the pillar group together with the peripheral wall portion, an upper end of the peripheral wall portion and an upper end of the partition portion are higher than the positions of the upper ends of the pillars; The pillar group includes: A micro-object capturing device that includes four or more of the pillars and captures micro-objects contained in a sample liquid injected into the recess at the central points of four adjacent pillars.

2. The micro object capturing device according to claim 1 , wherein the pair of side walls have notches at opposing positions into which both side portions of the partition are fitted.

3. the pillar group includes four or more of the pillars, At least four of the plurality of pillars constituting the pillar group are 3. A micro-object capturing device as described in claim 1 or 2, which has a rectangular shape in plan view, with the locations corresponding to the four corners of the rectangle being cut out in an arc shape.

4. the pillar group includes four or more of the pillars, At least four of the pillars constituting the pillar group are 3. A micro-object capturing device as described in claim 1 or 2, which has a square shape in plan view, with the four corners of the square being cut out in an arc shape.

5. At least one of the pillars constituting the pillar group is The micro-object capturing device according to claim 1 or 2, having an identifier on its surface.

6. The plurality of pillars constituting the pillar group include 3. The micro object capturing device according to claim 1 or 2, wherein the micro object capturing devices are arranged in a rectangular grid at preset intervals.

7. 3. The micro object capturing device according to claim 1, further comprising a recovery section into which the sample liquid injected into the recess flows.

8. The sample liquid injected into the recess further includes a recovery section into which the sample liquid flows, The tip member is A discharge flow path connecting the recess and a collection part, The partition portion is A plate-shaped base; 3. A micro-object capturing device as described in claim 1 or 2, further comprising a plug portion provided on one of the two end faces of the base for opening and closing the discharge flow path.

9. 3. A micro object capturing device as described in claim 1 or 2, further comprising a group partition portion above the pillar group, both sides of which are sandwiched by a pair of opposing side walls of the peripheral wall portion.

10. The tip member is A filter section disposed downstream of the pillar group and including a plurality of columnar sections; a collection section disposed downstream of the filter section, into which the sample liquid that has been injected into the recess and passed through the filter section flows, The filter unit includes: The micro-object capturing device according to claim 1 or 2, wherein the distance between the columnar portions is shorter than the distance between the pillars in the group of pillars.

11. The tip member is The micro object capturing device according to claim 10 , further comprising a communication channel connecting the recess and the filter section.

12. The partition portion is A plate-shaped base; A micro-object capturing device as described in claim 11 , further comprising a plug portion provided on one of the two end faces of the base for opening and closing the communicating flow path.

13. The pillar group includes: A plurality of subgroups each including four or more of the pillars, the pillars having a uniform shape and size; The plurality of pillars constituting the subgroup are They are arranged in a rectangular grid at preset intervals, The micro-object capturing device according to claim 1 or 2, wherein the pillar group includes sub-groups having pillars of different sizes.

14. a chip member including a pillar group consisting of a plurality of pillars protruding from a surface of a recess formed on a front side; a peripheral wall portion that is erected on an outer periphery of a surface of the tip member and surrounds the pillar group from all sides, The upper end of the peripheral wall portion is higher than the upper ends of the pillars, The pillar group includes: The method includes the step of detecting a micro-substance contained in a sample liquid injected into the recess, the micro-substance being captured at a central location between four adjacent pillars, and At least four of the plurality of pillars constituting the pillar group are A micro-object capture device that has a rectangular shape when viewed in a plane, with the areas corresponding to the four corners of the rectangle cut out in an arc shape.

15. a chip member including a pillar group consisting of four or more pillars protruding from a surface of a recess formed on a front side; a peripheral wall portion that is erected on an outer periphery of a surface of the tip member and surrounds the pillar group from all sides, The upper end of the peripheral wall portion is higher than the upper ends of the pillars, The pillar group includes: The pillars are arranged so that the central portions of the pillars are arranged adjacent to each other to capture minute objects contained in the sample liquid injected into the recess. At least one of the pillars constituting the pillar group is A micro-object capture device having an identifier on its surface.

16. a chip member including a pillar group consisting of four or more pillars protruding from a surface of a recess formed on a front side; a peripheral wall portion that is erected on an outer periphery of a surface of the tip member and surrounds the pillar group from all sides; a group partition portion, both sides of which are sandwiched between a pair of opposing side walls of the peripheral wall portion, above the pillar group; The upper end of the peripheral wall portion is higher than the upper ends of the pillars, The pillar group includes: A micro-object capturing device that captures micro-objects contained in a sample liquid injected into the recess at the central points of four adjacent pillars.

17. At least one of the tip member and the peripheral wall portion has An injection hole is provided for injecting a sample liquid, The pillar group includes: A plurality of subgroups each including four or more of the pillars and each having a uniform distance between the pillars; The pillar group includes: A micro-object capture device described in any one of claims 1, 14 to 16, wherein the distance between the pillars in the upstream sub-group is longer than the distance between the pillars in the downstream sub-group, based on the injection hole.

18. The peripheral wall portion has A micro-object capture device described in any one of claims 1, 14 to 16, wherein an adjustment hole for adjusting the amount of sample liquid injected into the recess is provided at a position higher than the upper end of each pillar in the group of pillars.

19. The tip member is A micro-object capturing device as described in any one of claims 1 and 14 to 16, wherein a surface including the group of pillars surrounded by the peripheral wall portion is subjected to a hydrophilic treatment.

20. A micro-object capturing device according to any one of claims 1 and 14 to 16, wherein an injection hole for injecting a sample liquid is provided in at least one of the tip member and the peripheral wall portion; a tubular injection port having one end attached to the injection hole; A micro-object capture system having an injection device including a reservoir connected to the other end of the injection port for storing sample liquid, and an electroosmotic pump for sending the sample liquid in the reservoir to the recess via the injection port.

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