JP2015047073A - Cell capturing device and cell capturing system - Google Patents

Abstract

PROBLEM TO BE SOLVED: To prevent air bubbles from staying in the vicinity of a filter inside a cell capture device.SOLUTION: A cell capture device 1 is formed to have chamfer portions where corners 127 (227) are chamfered by changing the shapes of a first protrusion 125 and a second protrusion 225 in the vicinity of the corners 127 (227) which are located at a large distance between a virtual liquid flow line S showing a liquid flow which connects bore holes 109 and 113 formed at end parts on the side of a cavity portion W of an introduction flow path to a bore hole 209 formed at an end part on the side of the cavity portion W of a discharge flow path and a prescribed point on an outer periphery of a filter region 33. This way, in the chamfered region, a distance between the prescribed point and an internal wall surface 330 of the cavity portion W is shorter than that prior to the change. This prevents air bubbles not only from staying in the vicinity of the filter 30, especially near the chamfered region, and also from being larger.

Description

  The present invention relates to a cell capture device that captures cells contained in a cell dispersion, and a cell capture system using the cell capture device.

  Cancer occupies the top cause of death in countries around the world, and more than 300,000 people die from cancer annually in Japan, and early detection and treatment thereof are desired. Most deaths due to cancer are due to recurrence of cancer metastasis. Recurrence of cancer metastasis occurs when cancer cells settle and infiltrate into the blood vessel wall of another organ tissue from the primary lesion via blood vessels or lymphatic vessels to form micrometastasis. Such cancer cells circulating in a human body through blood vessels or lymphatic vessels are called circulating tumor cells (hereinafter referred to as “CTC”).

Blood contains many blood cell components such as red blood cells, white blood cells, and platelets, and the number is said to be 3.5 to ⁹ × 10 ^{9 in} 1 mL of blood. On the other hand, since there are only a few CTCs, it is necessary to filter the blood in order to efficiently capture and detect CTC from blood cell components, and various studies have been made on this device. For example, in Patent Document 1, a microfluid having a polydimethylsiloxane (PDMS) upper member having a sample supply port above and below a nickel substrate having a fine through hole in a nickel substrate, and a lower member having a sample discharge port. A device is disclosed. Patent Document 2 discloses a biological component separation filter unit in which a lid member and a storage member sandwiching a filter are clamped and fixed as a device having improved dismantling properties.

JP 2011-163830 A JP 2003-70904 A

  However, in the devices shown in Patent Documents 1 and 2, when air is mixed into the flow path of the processing liquid used for blood and CTC capture processing, the measurement accuracy may be reduced. When air is mixed in the device and bubbles are accumulated, the bubbles block the flow path of blood or treatment liquid. Furthermore, when air bubbles stay in the vicinity of the filter in the device, it is considered that this also hinders the observation of the CTC captured by the filter.

  The present invention has been made in view of the above, and an object of the present invention is to provide a cell trapping device and a cell trapping system that can suppress bubbles from remaining in the vicinity of a filter in the cell trapping device.

  In order to achieve the above object, a cell capture device according to the present invention comprises a lid member having an introduction channel for introducing a test solution into the interior, and a discharge channel for discharging the test solution to the outside. A housing member having an inner wall surface formed by the lid member and the housing member between the introduction flow path and the discharge flow path. A filter that has a filter region in which a through-hole for passing a test solution is formed in the thickness direction and is supported by the lid member and the storage member outside the filter region of the filter; The shape of the said inner wall surface in planar view and the shape of the outer periphery of the said filter area | region are mutually different, It is characterized by the above-mentioned.

  As described above, the shape of the inner wall surface of the cavity in plan view is different from the shape of the outer periphery of the filter region, so that the outer side of the outer periphery of the filter region in the cavity portion and the inner wall surface of the cavity portion. The flow of the test solution in the region along the line does not become constant, and therefore it is possible to suppress the bubbles from staying between the outer periphery of the filter region near the filter and the inner wall surface of the cavity.

  Here, the outer periphery of the filter region is obtained by connecting the ends of the outermost through holes in the filter region. In plan view, the distance from the inner wall surface at a predetermined point on the outer periphery of the filter region is the predetermined point, the end of the introduction channel on the cavity side, and the cavity of the discharge channel. It is preferable that the aspect is changed based on the distance from the virtual liquid flow line indicating the flow of the test liquid connecting the end portion on the side.

  As described above, the virtual liquid flow line indicating the flow of the test liquid connecting the end portion on the cavity portion side of the introduction channel and the end portion on the cavity portion side of the discharge channel, and the outer periphery of the filter region By changing the distance between the predetermined point on the outer periphery of the filter region and the inner wall surface of the cavity based on the distance from the predetermined point, for example, the test is performed at a place far away from the virtual liquid flow line. In order to prevent stagnation, the flow of the liquid tends to be gentle, and in the region of the outer periphery of the filter region that is greatly separated from the virtual liquid flow line, each point on the outer periphery of the filter region and the inner wall surface of the cavity portion The distance between the predetermined point on the outer periphery of the filter region and the inner wall surface of the cavity can be adjusted more suitably according to the flow of the test solution, such as changing the shape of the inner wall surface so that the distance of It is possible to further suppress the retention of bubbles near the filter. It can be. The distance from the inner wall surface at a predetermined point on the outer periphery of the filter region and the distance from the virtual liquid flow line at the predetermined point both indicate the shortest distance.

  In addition, as a configuration that effectively exhibits the above-described operation, specifically, the filter region has a substantially square shape, and the end of the introduction flow path on the cavity portion side is the filter region in the lid member. Each of the four corners is provided at a position corresponding to two opposite corners, and the end of the discharge channel on the cavity portion side is a bottom surface of the cavity portion of the storage member, and the filter The inner wall surface of the cavity portion in plan view is provided in the center of the region corresponding to the region, and has a substantially rectangular shape corresponding to the filter region, and the cavity of the introduction flow path among the four corners of the filter region. The aspect which has a chamfering part in the area | region corresponding to two corners where the edge part by the side of a part is not provided is mentioned.

  At this time, it is more preferable that the chamfered portion has a curved surface.

  As described above, when the chamfered portion has a curved shape, it is possible to suppress bubbles from staying at the end of the chamfered portion on the inner wall surface of the cavity, and to further suppress the retention of bubbles near the filter. can do.

  Further, it is more preferable that the hollow portion has a curved surface from the top surface on the lid member side to the inner wall surface.

  By forming a curved surface from the top surface to the inner wall surface of the cavity as described above, it is possible to suppress bubbles from staying at the boundary between the top surface and the inner wall surface, and to further prevent bubbles from remaining in the vicinity of the filter. Can be suppressed.

  The cell capture system according to the present invention includes the above-described cell capture device, a test solution supply means for supplying a test solution to the introduction channel of the cell capture device, and an introduction channel of the cell capture device. A processing liquid supply means for supplying a processing liquid for processing the cells captured by the filter by passing through the filter, and a selection means for selecting a liquid to be supplied to the cell trapping device from the test liquid and the processing liquid And comprising.

  According to the cell capture system, the liquid to be supplied to the cell capture device is selected by the selection means, and the test liquid or the processing liquid is supplied to the cell capture device based on the result. Compared with the prior art, it is possible to efficiently perform the operation of capturing cells in the test solution.

  ADVANTAGE OF THE INVENTION According to this invention, the cell capture device and cell capture system which can suppress that a bubble stays in the filter vicinity in a cell capture device are provided.

It is a schematic perspective view explaining the structure of a cell trapping device. It is a schematic perspective view from the upper part about a lid member. It is a schematic perspective view from the lower part about a lid member. It is a figure explaining the internal structure of a cover member, and is an arrow line view of the VV cross section of FIG. It is a schematic perspective view from the upper side about a storage member. It is a schematic perspective view from the lower part about a storage member. It is a figure explaining the internal structure of a storage member, and is an arrow directional view of the VII-VII cross section of FIG. It is the perspective view and top view of a fixing member. It is a figure explaining the structure of a filter. It is a figure which shows the other example of the shape of the through-hole provided in a filter. It is a disassembled perspective view explaining the structure of a cell trapping device. It is the figure which showed typically the cross section before assembling a cover member, a filter, a sealing member, and a storage member, and is equivalent to XII-XII sectional drawing of FIG. It is a figure after assembling a cover member, a filter, a seal member, and a storage member, and is a figure corresponding to FIG. It is a figure which shows the mounting method of a fixing member. It is a block diagram explaining the structure of a cell trapping system. It is a schematic diagram explaining the structure of the cavity part inside the housing of a cell trapping device. It is a schematic diagram explaining the structure of the cavity part inside the housing of a cell trapping device, and is a figure corresponding to the arrow view of the XVII-XVII cross section of FIG. It is a figure explaining the modification of a structure of a cavity part.

  DESCRIPTION OF EMBODIMENTS Hereinafter, embodiments for carrying out the present invention will be described in detail with reference to the accompanying drawings. In the description of the drawings, the same elements are denoted by the same reference numerals, and redundant description is omitted.

(Cell capture device)
FIG. 1 is a schematic perspective view illustrating the configuration of a cell trapping device according to an embodiment of the present invention. A cell capturing device 1 shown in FIG. 1 has a function of capturing specific cells in a body fluid by a filter provided therein. As a particularly preferred example, a blood cell containing circulating cancer cells called CTC has a function of capturing red blood cells, platelets and white blood cells (hereinafter collectively referred to as “blood cell components”) while capturing CTC. Have. The cells captured by the filter are subjected to a staining process by introducing a washing solution, a staining solution, or the like into the inside, and then observed with a microscope or the like.

A cell capturing device 1 shown in FIG. 1 includes a housing 10 and a filter that is provided inside the housing 10 and allows a test solution to pass therethrough. Here, the test solution is a solution in which cells are dispersed. The housing 10 includes a lid member 100 and a storage member 200. Among them, the area of the region through hole of the filter is provided are 25mm ² ~400mm ^2. The area of the region through hole is ^provided, more preferably 25mm ² ~225mm ^2, more preferably 25 mm 2 100 mm ^2.

  Moreover, as a dimension and shape of the part enclosed by the lid member side guide part or the storage member side guide part which will be described later, from the viewpoint of workability and observability, the length of one side in a plan view is a substantially square with a length of 10 to 100 mm. Preferably there is. The length of one side is more preferably 15 to 70 mm, and still more preferably 20 to 30 mm.

  The lid member 100 and the storage member 200 are fixed to each other by being sandwiched by four fixing members 60 (60A to 60D) from both sides in the direction in which they are overlapped (the direction intersecting the filter). The four fixing members 60 are arranged at the peripheral edge of the housing 10 so as to surround the filter.

  As materials for the lid member 100 and the storage member 200, it is preferable to select materials having relatively high rigidity. Thereby, the deformation | transformation by the pressurization at the time of assembling and fixing the cell capture device 1 can be suppressed. However, if the rigidity is too high, the filter is damaged when stress is applied to the fitting portion. Here, the rigidity of the material can be expressed by, for example, Young's modulus. Therefore, the material of the lid member 100 and the storage member 200 is preferably 0.1 GPa to 100 GPa in terms of Young's modulus at room temperature, and more preferably 1 GPa to 10 GPa. The Young's modulus can be measured by a generally known Ewing method.

  Furthermore, the lid member 100 is preferably made of a material having translucency with respect to light having a wavelength used when detecting cells to be observed, particularly light in the visible light region. Examples of the material of the lid member 100 include, but are not limited to, glass, quartz glass, plastic (particularly acrylic resin), and a polymer such as polydimethylsiloxane. The lid member 100 and the storage member 200 are not necessarily made of the same material, but are preferably the same material from the viewpoint that stress applied to the fitting portion during assembly is applied equally to the lid member side and the storage member side. The material of the lid member 100 and the storage member 200 is preferably a low-autofluorescent acrylic resin, and particularly preferably polymethyl methacrylate because the device can be mass-produced. In general, when observing cancer cells or the like, after subjecting the target cells to a staining treatment with a fluorescent reagent, irradiation with ultraviolet or visible light having a wavelength of 300 to 800 nm is performed for fluorescence observation. Do. Therefore, it is preferable to select a material having low autofluorescence for the lid member 100 so that the material itself does not emit fluorescence when irradiated with light in the wavelength range (this is referred to as “low autofluorescence”). . In general, an organic polymer having an aromatic ring, such as a resin such as polystyrene or polycarbonate, has strong autofluorescence and is often not suitable for the above purpose.

  The lid member 100 includes introduction flow channels 101 and 102 for introducing a test liquid containing cells to be observed and a treatment liquid for staining the cells captured on the filter. The storage member 200 has a discharge channel 201 for discharging the test liquid from the inside to the outside. In the following description, the direction in which the introduction flow paths 101 and 102 are attached is the X-axis direction, the horizontal direction orthogonal to the X-axis is the Y-axis direction, and the vertical direction orthogonal to the X-axis and the Y-axis is Z Axial direction.

  Next, the configuration of the cell trapping device 1 will be described in detail. First, the configuration of the lid member 100, the storage member 200, the fixing member 60, and the filter will be described, and then the cell trapping device in which these are assembled will be described.

(Cover member)
First, the lid member 100 will be described. 2 to 4 are diagrams illustrating the configuration of the lid member 100, FIG. 2 is a schematic perspective view from above of the lid member 100, and FIG. 3 is a schematic perspective view of the lid member 100 from below. FIG. 4 is an arrow view of the VV cross section of FIG.

  As shown in FIGS. 2 to 4, the lid member 100 has two main body parts 110 that are substantially square plate members so as to protrude outward along the X-axis direction from the side edges of the main body part 110. The flow path parts 105 and 106 are attached. The flow path portions 105 and 106 are attached at positions shifted from the center along the Y-axis direction so that they are located at different positions when the lid member 100 is viewed from the X-axis direction. Here, the flow path unit 106 shown in FIG. 4 will be described, but the flow path unit 105 has the same configuration.

  The lid member 100 is provided with an introduction region 120 formed of a concave gap for allowing the test liquid to pass through the filter. The introduction region 120 includes a region (filter region) in which a through-hole is provided in the filter attached between the lid member 100 and the storage member 200 when the cell trapping device 1 is viewed from above. Provided above the filter area. The introduction flow paths 101 and 102 are connected to the corners of the introduction region 120 through connection holes 109 and 113. The introduction region 120 is a space for guiding the test liquid introduced from the introduction flow channels 101 and 102 to the through hole of the filter.

  Here, the introduction channel 102 will be further described with reference to FIG. The flow path portion 106 is provided with an introduction flow path 102 extending along the X axis. The introduction channel 102 includes an introduction port 102A, a channel 102B extending inward from the introduction port 102A, and a channel 102C connected to the channel 102B and having a smaller diameter than the channel 102B. Further, a perforation 111 is provided between the flow path 102B and the flow path 102C so as to penetrate the flow path portion 106 in the vertical direction (Z-axis direction) (the perforation 107 is provided for the flow path portion 105). . A valve provided with a perforation functioning as a flow path opening / closing valve (flow path opening / closing mechanism) is inserted into the perforation 111. Thereby, the flow path 102B is opened and closed by this valve. Further, a perforation 113 that connects the introduction region 120 and the introduction channel 102 is provided at the end of the introduction channel 102 on the main body 110 side so as to communicate with the introduction channel 102. Thereby, the liquid (test liquid or treatment liquid) introduced from the introduction port 102A is sent to the introduction region 120 through the flow path 102B, the flow path 102C, and the perforations 113.

  A first protrusion 125 for preventing the liquid introduced into the introduction region 120 from leaking outside is provided at the periphery of the introduction region 120 so as to extend downward from the main body 110. When the lid member 100 is viewed from below, the first projecting portion 125 has a substantially quadrangular shape in the XY plane as shown in FIG. 3, and the introduction flow passages 101 and 102 like two wind turbines. Only the region connected to the perforations 109 and 113 leading to the shape is formed to protrude from the outer periphery of the square shape. Further, a first fitting portion 130 formed of a quadrangular convex portion is formed so as to be separated from the first protruding portion 125 and surround the periphery of the first protruding portion 125. The length of one side of the first fitting portion 130 in the X-axis direction or the Y-axis direction is preferably 8 to 30 mm, more preferably 10 to 25 mm, from the viewpoint of the strength of the lid member 100 and the fitting property of the filter 30. Preferably, 15-20 mm is more preferable. Moreover, 1-5 mm is preferable, as for the width | variety of the convex part which comprises each edge | side of the 1st fitting part 130, 1-3 mm is more preferable, and 1.5-2 mm is still more preferable. As shown in FIG. 4, the height of the first fitting portion 130 is higher than that of the first protruding portion 125. Furthermore, an alignment wall 135 that protrudes downward (upward in FIG. 3) is provided only on the outer edge of the main body 110 that extends along the Y-axis direction. This is used for aligning and aligning the lid member 100 and the storage member 200 when assembling the cell trapping device 1.

  The shapes of the introduction region 120 and the first protrusion 125 of the lid member 100 will be described in detail. In the introduction region 120 partitioned by the first protrusion 125, the perforations 109 and 113 leading to the introduction flow channels 101 and 102 are formed. In two corners 127 different from the two corners provided with the region to be connected, a so-called chamfered shape is formed on the side surface constituting the inner wall surface of the introduction region 120 in the substantially quadrangular first protrusion 125. Has been. Further, the top surface of the internal space including the introduction region 120 (the surface on the lower side in FIG. 3) and the side surface of the first protrusion 125 that constitutes the inner wall surface of the introduction region 120 are connected by a curved surface. Yes. These structures are structures for preventing bubbles from staying in the hollow portion inside the housing 10 including the introduction region 120, but will be described in detail later.

  Returning to FIG. 2, in the body part 110 having a substantially square shape, the peripheral parts of the four sides of the surface that becomes the upper surface when assembled are each started from the vicinity of the end part (corner part) and along the peripheral part. Lid member side guide portions 151 (151A to 151D) extending in a counterclockwise direction in plan view are provided. The lid member side guide portion 151 is a protrusion that protrudes upward from the lid member 100, and defines a direction in which the fixing member 60 slides when a claw of the fixing member 60 described later engages with this. . Among the four peripheral portions, the peripheral portion in which the flow channel portions 105 and 106 are attached to the side edges, among the peripheral portions divided by the flow channel portions 105 and 106, the longer distance side is the lid member side. Guide portions 151A and 151B are provided, respectively. That is, the lid member side guide portion 151A and the lid member side guide portion 151B are provided at positions that are diagonal with respect to the center of the main body portion 110 in plan view. On the other hand, in the remaining peripheral part to which the flow path parts 105 and 106 are not attached, when the main body part 110 is viewed from the Y-axis direction, the flow path part located on the front side of the flow path parts 105 and 106 is not. The lid member side guide portions 151C and 151D are respectively provided on the attached side. That is, the lid member side guide portion 151C and the lid member side guide portion 151D are provided at positions that are diagonal with respect to the center of the main body portion 110 in plan view. As shown in FIG. 2, the stepped portion between the starting point of the lid member side guide portion 151 and the upper surface of the main body 110 is chamfered obliquely.

  The lid member side guide portions 151 (151A to 151D) extend to the substantially central portion starting from the vicinity of the end of the main body portion 110 at each peripheral edge of the main body portion 110. From the substantially central portion, the lid member side stopper 152 is provided. (152A to 152D) are continuously provided and extend along the peripheral edge. The lid member side stoppers 152A to 152D are protrusions having a height higher than that of the lid member side guide portions 151A to 151D, and the limit position where the fixing member 60 described later slides along the lid member side guide portions 151A to 151D. It prescribes. The lid member side stoppers 152A to 152D extend to the vicinity of the end portion on the opposite side to the starting point of the lid member side guide portions 151A to 151D at the respective peripheral portions. The lid member side stoppers 152A and 152B at the peripheral edge to which the flow path portions 105 and 106 are attached are partially integrated with the root portions of the flow path portions 105 and 106.

  Note that the width in the X-axis direction of the lid member side guide portions 151A and 151B and the lid member side stoppers 152A and 152B is larger than the width in the Y axis direction of the lid member side guide portions 151C and 151D and the lid member side stoppers 152C and 152D. It has been enlarged. The reason for this will be described later.

(Storage member)
Next, the storage member 200 will be described. 5 to 7 are diagrams illustrating the configuration of the storage member 200, FIG. 5 is a schematic perspective view of the storage member 200 from above, and FIG. 6 is a schematic perspective view of the storage member 200 from below. FIG. 7 is a sectional view taken along the line VII-VII in FIG.

  As illustrated in FIGS. 5 to 7, the storage member 200 has a flow path portion 205 that protrudes outward along the Y-axis direction from the side edge of the main body portion 210 with respect to the main body portion 210 formed of a substantially square plate member. Is attached. The channel part 205 is attached to the center of the side edge of the main body part 210.

  The storage member 200 is provided with a discharge region 220 formed of a concave gap through which the test liquid that has passed through the filter passes. The discharge region 220 is below the region where the filter is attached so as to include the region (filter region) in which the through-hole is provided in the filter attached to the storage member 200 when the cell trapping device 1 is viewed from above. Provided. The discharge area 220 is connected to the discharge flow path 201 via the connection perforations 209, and becomes a space for discharging the test liquid to the outside through the discharge flow path 201.

  Here, the discharge channel 201 will be further described with reference to FIG. The flow path portion 205 is provided with a discharge flow path 201 extending along the Y axis. The discharge flow channel 201 includes a discharge port 201A, a flow channel 201B extending inward from the discharge port 201A, and a flow channel 201C connected to the flow channel 201B and having a smaller diameter than the flow channel 201B. Further, a perforation 207 that penetrates the flow channel portion 201 in the vertical direction (Z-axis direction) is provided on the flow channel 201C. A valve provided with a hole that functions as a flow path opening / closing valve is inserted into the hole 207. Thereby, the flow path 201C is opened and closed by this valve. Further, a perforation 209 that connects the discharge region 220 and the discharge channel 201 is provided at the end of the discharge channel 201 on the main body 210 side so as to communicate with the discharge channel 201. Thereby, the liquid (test liquid or processing liquid) that has reached the discharge region 210 via the filter is sent to the discharge port 201A via the perforation 209, the flow path 201C, and the flow path 201B.

  A second projecting portion 225 is provided at the periphery of the discharge region 220 so as to extend upward from the main body portion 210 to suppress height variation in the XY plane of the filter and to flatten the filter. When the storage member 200 is viewed from above, the second protrusion 225 has a substantially square shape and a shape of two windmills in the XY plane, as shown in FIG. This is a shape corresponding to the first protrusion 125 of the lid member 100. In addition, a second fitting portion 230 formed of a quadrangular concave portion is formed so as to be separated from the second protruding portion 225 and surround the periphery of the second protruding portion 225. The second fitting portion 230 is provided so as to correspond to the first fitting portion 130 of the lid member 100.

  In the region between the second projecting portion 225 and the second fitting portion 230, in which the portion corresponding to the wing in the “shape of two wind turbines” is not provided, Two bottomed holes (holes) 240 are provided at positions diagonal to each other. The bottomed hole 240 is used for positioning the storage member 200 and the filter 30 as described later. The bottomed hole 240 may be a through hole that penetrates the storage member. In the present embodiment, an example in which the bottomed hole 240 is provided on the storage member 200 side is described, but the bottomed hole 240 may be provided on the lid member 100 side.

  The difference in size between the second fitting portion 230 of the storage member 200 and the first fitting portion 130 of the lid member 100 is preferably determined in consideration of the thickness of the filter 30. The difference between the inside of the second fitting portion 230 and the inside of the first fitting portion 130 (difference inside the fitting portion as viewed from the center of the member) is preferably 0.005 to 0.3 mm, and 0.005 to 0.2 mm is more preferable, and 0.005-0.1 mm is still more preferable. When the difference between the inside of the second fitting portion 230 and the inside of the first fitting portion 130 is smaller than the thickness of the filter, there is no gap than the thickness of the filter, but the lid member and the storage member are made of plastic. It is preferable because it is relatively elastic and the fixing force when the filter is fitted increases.

  In addition, the storage member 200 has a direction orthogonal to the extending direction (X-axis direction) of the outer edge of the main body 210 apart from the second fitting portion 230, that is, the Y-axis, that is, the Y-axis. The alignment groove 235 is provided only on the side extending along the direction. This is used for aligning and aligning the lid member 100 and the storage member 200 when assembling the cell trapping device 1. Further, in the same direction (X-axis direction) as the direction in which the outer edge where the flow path portion 201 is provided, out of the outer edges of the main body 210, a cover portion 250 that extends upward and covers the side portion of the lid member 100 is provided. It has been.

  Of the discharge region 220 delimited by the second projecting portion 225, when combined with the lid member 100, the region connected to the perforations 109 and 113 leading to the introduction channels 101 and 102 is formed on the lid member 100 side. The two corners 227 different from the two corners have a so-called chamfered shape on the side surface constituting the inner wall surface of the introduction region 120 in the first projection 125 having a substantially square shape.

  As shown in FIG. 6, in the body portion 210 having a substantially square shape, the periphery of the four sides of the surface that becomes the bottom surface (surface exposed to the front) when assembled is the vicinity of the end portion (corner portion). And storage member side guide portions 251 (251A to 251D) extending along the respective peripheral edge portions. The storage member side guide portions 251A to 251D are protrusions protruding downward (upward in FIG. 6) of the storage member 200, and the fixing member 60 slides by engaging the claws of the fixing member 60 described later. It defines the direction to be performed. The storage member side guide portions 251 </ b> A to 251 </ b> D are provided at positions corresponding to the cover member side guide portions 151 </ b> A to 151 </ b> D in the thickness direction of the housing 10 when the storage member 200 and the lid member 100 are assembled. Yes. For example, at the peripheral edge to which the flow path portion 205 is attached, when the main body 210 in FIG. 251C is provided. As shown in FIG. 6, the stepped portion between the starting point of the lid member side guide portions 151 </ b> A to 151 </ b> D and the upper surface of the main body portion 110 is chamfered obliquely.

  The storage member side guide portions 251 (251A to 251D) extend to the substantially central portion starting from the vicinity of the end of the main body portion 210 at each peripheral edge of the main body portion 210, and from the substantially central portion, the storage member side stopper 252. (252A to 252D) are continuously provided and extend along the peripheral edge. The storage member side stoppers 252A to 252D are protrusions having a height higher than the storage member side guide portions 251A to 251D, and the limit position where the fixing member 60 described later slides along the storage member side guide portions 251A to 251D. In addition, the cell capture device 1 functions as a leg of the cell capture device 1 during use and observation (see FIG. 7). The storage member side stoppers 252A to 252D extend to the vicinity of the end portion on the opposite side to the starting point of the storage member side stoppers 252A to 252D at the respective peripheral portions. A part of the storage member side stopper 252 </ b> C at the peripheral edge to which the flow path part 205 is attached is integrated with the base part of the flow path part 205.

  When the lid member 100 and the storage member 200 are made of plastic such as acrylic resin, they can be manufactured by injection molding. However, the manufacturing method of the lid member 100 and the storage member 200 is not limited to the above.

(Fixing member)
Next, the fixing member 60 will be described. 8A is a perspective view of the fixing member, and FIG. 8B is a front view of the fixing member 60. FIG. The fixing member 60 is an integrally formed member having a shape in which one side surface of a cylindrical body having a substantially rectangular cross section is opened (that is, a U-shaped cross section), and the lid member 100 and the storage member 200 are attached to each other. These are sandwiched and fixed so as to be pressed from both sides in the direction in which they are superposed (direction intersecting with the filter).

  The fixing member 60 has claws 61 and 61 that protrude from each other in a direction in which the lid member 100 and the storage member 200 should be sandwiched at the U-shaped inner surface portion and the U-shaped tip portion. 61 extends in a direction corresponding to the height of the “substantially rectangular cylindrical body”. That is, the claws 61 and 61 extend in a direction along the peripheral edge portions of the lid member 100 and the storage member 200 when the fixing member 60 is disposed at the peripheral edge portion of the housing 10, and the lid member-side guide portion 151 and the storage member. By being engaged with the side guide portion 251, it is possible to slide along the lid member side guide portion 151 and the storage member side guide portion 251. Here, the length of the fixing member 60 in the sliding direction is preferably 3 to 20 mm, more preferably 4 to 15 mm, and still more preferably 5 to 10 mm from the viewpoint of pressing more uniformly. The length is preferably 0.2 to 0.4 as a ratio with respect to the length of one side of the peripheral portion of the casing 10 to which the fixing member 60 is mounted.

  From the viewpoint of pressing and fixing the peripheral edge of the housing 10 with a uniform load, it is preferable to use the same material as the plurality of fixing members 60 to be used. For example, it is preferable to use those having the same shape and size, or those having small variations in pressing force.

The material of the fixing member 60 is preferably a material having moderate elasticity (Young's modulus of 2 to 5 GPa), excellent tensile strength (500 Kg / cm ² or more), and an elongation of 10 to 100%. Polycarbonate or polyacetal resin is preferred.

(filter)
Next, the filter 30 provided between the lid member 100 and the storage member 200 will be described with reference to FIG.

  As shown in FIG. 9, the filter 30 is configured to include a filter region 33 in which a plurality of through holes 32 are formed in the thickness direction in the sheet 31 and allow the test liquid to pass therethrough. In FIG. 9, the shape of the through hole 32 on the surface of the sheet 31 is a wave shape. The corrugated through-hole 32 is formed by connecting a plurality of perforations having a rectangular or rounded rectangular shape on the sheet surface at a predetermined crossing angle between the ends. In the filter 30 of FIG. 9, the wave shape of the through hole 32 is formed along the X-axis direction.

  The sheet 31 has a substantially square shape, but is provided with square-shaped cut portions 34 at four corners. The outer side from the region where the cut portion 34 is formed is a bent region 35. Further, an alignment hole 36 which is a through hole different from the through hole 32 in the filter region 33 is formed at a position outside the filter region 33 and inside the outer edge of the filter 30. Two alignment holes 36 are formed at diagonal positions of the substantially square of the sheet 31. The folding region 35 and the alignment hole 36 are used when the filter 30 is attached to the cell trapping device 1, which will be described later.

  The material of the sheet 31 is preferably composed mainly of metal. Here, the main component refers to the component having the largest proportion of the material forming the sheet 31. By using a metal as the main component of the material of the filter 30 sheet, cells can be separated and concentrated with high separation accuracy with little variation in the size of the through-holes. In addition, since metal is more rigid than other materials such as plastic, its size or shape is easily maintained even when a force is applied from the outside. For this reason, it is considered that blood components (particularly white blood cells) that are slightly larger than the diameter of the through-holes are deformed and allowed to pass, and separation and concentration with high accuracy are possible. Among leukocytes, there are cells having the same size as the cells to be separated and concentrated, and only the target cells may not be distinguished at a high concentration only by the difference in size. However, since leukocytes are more deformable than the above cells, they can pass through pores smaller than themselves by external force such as suction or pressurization, and the cells and leukocytes are separated or concentrated. It will be possible. Here, “concentration” means that the existence ratio of the number of cells to be separated and concentrated with respect to the number of white blood cells increases before and after the test solution is passed through the cell capture device.

  Examples of the metal material used for the sheet 31 include gold, silver, copper, aluminum, tungsten, nickel, chromium, and alloys of these metals, but are not limited thereto. In addition, the metal may be used alone, or may be used as an alloy with another metal or a metal oxide in order to impart functionality. From the viewpoint of price or availability, it is preferable to use nickel, copper, gold, and a metal containing these as a main component, and it is particularly preferable to use a metal containing nickel as a main component. Moreover, when the sheet | seat 31 is formed from the material which has nickel as a main component, it is preferable that the surface of nickel is gold-plated. Since the surface of the filter can be prevented from being oxidized by gold plating, the adherence of cells and blood cell components to the filter becomes uniform, and the reproducibility of data can be improved.

  The thickness of the filter 30 is preferably 3 μm to 100 μm. When the film thickness is in the above range, the filter is easy to handle and suitable for precision processing. Further, the difference in level of the plane of the filter region of the filter is preferably 16 μm or less as the difference between the maximum and minimum focal lengths when a total of five locations on the end and center of the filter surface are observed with a microscope.

The size of the filter 30 depends on the size of the cell trapping device 1, but the size of the filter region 33 in which the through hole 32 is provided in the filter 30 and allows the test solution to pass is 25 mm ^2. ～400Mm ² are ^preferred, more preferably 25mm ² ~225mm ^2, more preferably 25 mm 2 100 mm ^2. When the size of the filter region 33 exceeds 400 mm ² , the dead space increases. Moreover, when it is less than 25 mm ² , the processing time becomes long. The size of the filter region 33 corresponds to the central portion 120A of the introduction region 120 of the lid member 100 shown in FIG. 3 and the central portion 220A of the discharge region 220 of the storage member 200 shown in FIG. That is, the filter region 33 of the filter 30 is attached at a position corresponding to the above-described central portions 120A and 220A.

  As for the shape of the through hole 32 provided in the filter 30, as shown in FIG. 9, the end of the perforation having a wave shape, that is, two rectangular shapes or rounded rectangular shapes (rectangles with rounded corners). It can be set as the shape formed by connecting each other at a predetermined angle. In the case of the through hole 32 having a rectangular shape or a rounded rectangular shape (hereinafter collectively referred to as “substantially rectangular shape”), the range of the length of the short side of the substantially rectangular shape is about 5 It can be set to 0.0 μm to 15.0 μm. On the other hand, the length of the long side can be appropriately changed according to the size of the filter, and the range is approximately 10 μm to 5 mm. In addition, the hole diameter of the through-hole 32 provided in the filter 30 is changed according to the size of CTC to be captured. Here, the hole diameter of the through hole refers to the diameter of the largest sphere that can pass through each through hole 32. For example, the diameter of the through hole 32 in FIG. 9 can be the length of the short side of the perforation.

  In addition, about the shape of the through-hole 32, although FIG. 9 demonstrated the case of the waveform, it can be changed into another shape. As a typical modification of the shape of the through hole, as shown in FIG. 10, (A) a circle, (B) a rectangle, (C) a rounded rectangle (the corner of the end is rounded), (D) Rounded rectangle (rectangular short side is arc-shaped), (E) Wave shape (three rounded rectangular perforations are connected to each other at the end), (F) Circular wave shape (five semicircles are alternately facing each other and connected at the end). Note that the shape of the through hole is not limited to the example shown in FIG. 10. For example, the shape of the through hole may be a wave shape in which four or more rounded rectangular perforations are connected, or a rectangular shape that is not rounded but rounded. A through-hole having a shape in which a plurality of perforations are connected may be used.

  As a method for manufacturing a filter capable of precisely forming the through hole 32 having a small hole diameter on the filter 30, for example, there is a method using metal plating using a photoresist. Specifically, in this manufacturing method, a step of laminating a photoresist on the metal foil, a step of exposing a photomask having a light-transmitting portion similar to the shape of the through-hole on the photoresist, and exposing, Developing a photoresist pattern by removing uncured portions of the photoresist, forming a metal plating pattern lower than the height of the photoresist pattern by metal plating between the photoresist patterns, and a metal foil. A step of obtaining a structure composed of a metal plating pattern and a photoresist pattern by removing by chemical dissolution, and a metal plating pattern having a through hole 32 corresponding to the light transmitting portion by removing the photoresist pattern from the structure ( Obtaining a filter). However, the manufacturing method of the filter 30 is not limited to the above method.

(Assembly method of cell capture device)
Next, a method for assembling the cell trapping device 1 will be described with reference to FIGS. 11 is an exploded perspective view illustrating the configuration of the cell trapping device 1. FIG. 12 is a schematic diagram illustrating a cross section before the lid member 100, the filter 30, the seal member 40 (gasket), and the storage member 200 are assembled. FIG. 13 is a view corresponding to FIG. 12 after assembling the lid member 100, the filter 30, the seal member 40, and the storage member 200. FIG. FIG. 4 is a view showing a method for mounting a fixing member.

  As an outline of the assembly method of the cell trapping device 1, as shown in FIG. 11, after the storage member 200, the seal member 40B, the filter 30, the seal member 40A, and the lid member 100 are overlapped in this order, FIGS. As shown in FIG. 3, the storage member 200 and the lid member 100 are fitted. Then, by attaching the fixing member 60 to the housing 10 formed by fitting the lid member 100 and the storage member 200 as shown in FIG. 14, the lid member 100 and the storage member 200 are fixed to each other. At the same time, the assembly of the cell trapping device 1 is completed.

  First, as shown in FIG. 11, the lid member 100, the lid member side seal member 40 </ b> A, the filter 30, the storage member side seal member 40 </ b> B, and the storage member 200 are stacked in this order from top to bottom along the Z axis. The cell capture device 1 is assembled. At this time, since the lid member 100 and the storage member 200 are oriented so that the pair of alignment walls 135 and the alignment groove 235 face each other, the flow path portions 105 and 106 of the lid member 100 are arranged in the X-axis direction. The extending direction is such that the flow path portion 205 of the storage member 200 extends in the Y-axis direction. Thereby, it becomes the structure which the flow-path parts 105,106,205 protrude in a mutually different direction.

  Here, the seal member 40 (40A, 40B) is sandwiched between the first protrusion 125 of the lid member 100 and the filter 30 and between the second protrusion 225 of the storage member 200 and the filter 30. It is a member. The seal member 40 is disposed in all areas where the first protrusion 125 and the second protrusion 225 exist, and the center is punched into the shape of the filter area 33 so as not to cover the filter area 33. It has a ring shape. The material of the seal member 40 is not particularly limited as long as it has elasticity and can maintain the liquid-tightness of the housing 10. For example, silicone rubber is used. The thickness of the seal member 40 is preferably 0.05 μm to 0.3 μm.

  After these are assembled, valves 401, 402, and 403 that function as flow path opening and closing valves are inserted into the perforations 107, 111, and 207, respectively. These valves 401, 402, and 403 are each provided with flow passage holes 401 A, 402 A, and 403 A that are perforations extending in the horizontal direction, and when the valves are inserted into the perforations 107, 111, and 207, the flow passage holes of the valves are provided. Correspond to the introduction channels 101 and 102 and the discharge channel 201, respectively. And opening and closing of a flow-path hole and an introduction flow path (or discharge flow path) can be switched by rotating a valve. In use, the stoppers 411, 412, and 413 are inserted into the perforations 109, 113, and 209, respectively, to prevent leakage of the liquid and guide the introduction and discharge of the liquid in an appropriate direction.

  Next, an internal configuration when the cell capturing device 1 is assembled will be described with reference to FIGS. 12 and 13. FIG. 12 is a view schematically showing a cross section before assembling the lid member 100, the filter 30, and the storage member 200, but the flow path portion 205 is not described. FIG. 13 is a view after assembly.

  First, as illustrated in FIG. 12, the storage member side seal member 40 </ b> B is disposed at a position above the second protrusion 225 of the storage member 200.

  Next, the filter 30 is disposed at a position above the discharge region 220, the second protrusion 225, and the second fitting portion 230 of the storage member 200. When placing the filter 30, a rod-shaped jig is passed through the alignment holes 36 and 36 of the filter 30, and the end portions of the jig are inserted into the bottomed holes 240 and 240 of the storage member 200, so that The filter 30 can be aligned. Alternatively, the filter 30 can be easily aligned by allowing the jig to stand in the bottomed holes 240 and 240 of the storage member 200 and passing the alignment hole 36 of the filter 30 therethrough. Thus, the position corresponding to the bottomed hole 240 and the alignment hole 36 in the storage member side seal member 40B is cut out so that the filter 30 can be aligned using the jig. (See FIG. 11).

  Next, the lid member side seal member 40 </ b> A is disposed at a position above the filter 30. And the cover member 100 is arrange | positioned in the position where the 1st protrusion part 125 and the 2nd protrusion part 225 respond | correspond, and the 1st fitting part 130 and the 2nd fitting part 230 correspond in the position corresponding. Arrange them and bring them close together in the vertical direction.

  Then, as shown in FIG. 13, the lid member 100 and the first projecting portion 125 and the second projecting portion 225 reach the position where the filter 30 is sandwiched through the lid member side seal member 40A and the storage member side seal member 40B, respectively. The storage member 200 can be brought close to each other. At this time, the first fitting portion 130 and the second fitting portion 230 stop in a state where a part thereof is fitted. At this time, the filter 30 is supported by being sandwiched between the first projecting portion 125 and the second projecting portion 225, and further on the outer side, the first fitting portion 130 and the second fitting portion 230 are supported. Between. As a result, the folding region 35 of the filter 30 is bent downward mainly by pushing down the lid member 100. And by being pinched so that it may be pressed from the center toward the outside from the center part inside the filter 30, a force pulled from the center toward the periphery is applied, and the filter 30 and the lid member 100 in a state where tension is applied. It is fitted between the storage member 200. Thereby, the variation in height of the filter surface is suppressed, and the flatness of the filter 30 becomes excellent.

  In addition, when the force at the time of pinching the filter 30 by the cover member 100 and the storage member 200 becomes non-uniform | heterogenous, there exists a possibility that distortion may arise and a filter 30 may wrinkle or may sag. Therefore, it is preferable that the second fitting portion 230 of the storage member 200 and the first fitting portion 130 of the lid member 100 are provided on the entire outer edge side of the filter 30 and are fitted so as to sandwich the filter 30 uniformly. However, since the filter 30 is sandwiched between the first protrusion 125 and the second protrusion 225 at this time, a slight tension may be applied in the circumferential direction to cause wrinkles. It is preferable to provide a notch 34 to create a wrinkle relief.

  In addition, after the storage member 200 and the filter 30 are aligned using the jig, the filter 30 is aligned with the storage member 200, but the position is not fixed. For this reason, until the filter 30 is finally fixed inside the housing 10, there is no physical obstacle to the pulling force or the like and the position change can be handled. When fixed between them, wrinkles and the like are less likely to occur.

  Next, a method for mounting the fixing member 60 will be described. As shown in FIG. 14, with respect to the starting point of the lid member side guide portion 151C and the storage member side guide portion 251C corresponding thereto, the fixing member 60C is placed with the U-shaped open portion facing the housing 10 side. The claws 61 and 61 are engaged with the lid member side guide portion 151C and the storage member side guide portion 251C, respectively, by approaching from the X-axis direction. Then, the mounting of the fixing member 60C is completed by sliding the fixing member 60C in the X-axis direction. At this time, the fixing member 60C may be slid until the side portion of the fixing member 60C contacts the side portion of the lid member side stopper 152C and the side portion of the storage member side stopper 252C. Good.

  Similarly, in order to attach the fixing member 60B to the housing 10, the fixing member 60B is U-shaped with respect to the lid member side guide portion 151B and the end portion that is the starting point of the storage member side guide portion 251B corresponding thereto. The claw 61, 61 is engaged with the lid member side guide portion 151B and the storage member side guide portion 251B by approaching from the Y-axis direction in a state where the opening portion is directed to the housing 10 side. Then, the mounting of the fixing member 60B is completed by sliding the fixing member 60B in the Y-axis direction.

  Focusing on the distance from the outer edge of the housing 10 to the lid member side guide portions 151C and 151B, the distance from the outer edge of the housing 10 to the lid member side guide portion 151C by the thickness of the cover portion 250 of the storage member 200. However, it is larger than the distance from the outer edge of the housing | casing 10 to the cover member side guide part 151B. As described above, the width in the X axis direction of the lid member side guide portion 151B and the lid member side stopper 152B is larger than the width in the Y axis direction of the lid member side guide portion 151C and the lid member side stopper 152C. The distance between the outer edge of the housing 10 and the portion where the claw 61 of the fixing member 60 of the lid member side guide portions 151C and 151B engages is equal. That is, the fixing member 60 having the same shape and the same size can be attached to each peripheral portion of the housing 10. The use of the fixing member 60 having the same shape and size as described above means that the same fixing member 60 having the same pressing force can be used, and the peripheral portion of the housing 10 is pressed with a uniform load. From the viewpoint of fixing.

  Although not shown in FIG. 14, the fixing members 60 </ b> A and 60 </ b> D can be attached to the housing 10 in the same manner as described above. By mounting the four fixing members 60 </ b> A to 60 </ b> D in this way, the fixing member 60 can be disposed on the peripheral edge of the housing 10 so as to surround the filter 30.

  As described above, the cell capturing device according to the embodiment of the present invention has been described by taking the configuration in which the lid member and the storage member are fixed by the fixing member as an example, but is not limited to this configuration, and the lid member and the storage member are welded. It is good also as a structure to join.

(Cell capture system)
Next, a cell capture system using the cell capture device 1 will be described. FIG. 15 is a block diagram illustrating the configuration of the cell trapping system. As shown in FIG. 15, the cell trapping system 1 </ b> A stores a cell trapping device 1 and a test solution and a treatment solution supplied via two introduction channels included in the cell trapping device 1, respectively. A liquid supply container 51 (test liquid supply means) and a processing liquid supply container 52 (processing liquid supply means), and a test liquid supply channel 53 (test) connecting the test liquid supply container 51 and the cell trapping device 1 Liquid supply means), a treatment liquid supply flow path 54 (treatment liquid supply means) for connecting the treatment liquid supply container 52 and the cell capture device 1, and a discharge flow path included in the cell capture device 1. With respect to the cell trapping device 1, a drainage flow channel 55 for flowing the drainage discharged to the outside from the drainage flow channel, a drainage recovery container 56 connected to the drainage flow channel 55 and collecting the drainage, The liquid to be supplied is selected from the test liquid and the processing liquid, and the flow path A control unit 57 for controlling such Toggles (selecting means), configured to include a.

  The control of the switching of the flow path and the like is performed by the control unit 57. As a specific method of switching the flow path, for example, there is a method of opening and closing a flow path opening / closing valve of the cell trapping device 1. In addition, the liquid feeding in the cell trapping system 1A can be performed, for example, by attaching a peristaltic pump or the like to the flow path. Moreover, although the example which has one type of process liquid was shown, it is set as the structure which provides the some process liquid supply container 52 and controls which process liquid is supplied with respect to the cell capture device 1 by the control part 57. FIG. You can also.

  As described above, in the cell trapping system 1A, the controller 57 selects the liquid to be supplied to the cell trapping device 1 and supplies the test liquid or the processing liquid to the cell trapping device 1 based on the result. By having the above, it becomes possible to efficiently advance the capturing operation of the cells in the test solution as compared with the conventional case. Further, in the cell trapping system 1A using the cell trapping device, after the cell trapping operation, the cell trapping device can be directly placed on the stage of the microscope without being disassembled and can be observed with good workability. .

(Operation in cell capture device)
Here, with reference to FIG. 16 and FIG. 17, the internal configuration of the casing 10 which is a characteristic part of the cell trapping device 1 described in the above embodiment will be described, and the operation thereof will be described. FIG. 16 is a view of a part of the inside of the cell trapping device 1 according to the present embodiment as seen from above, and is a diagram schematically showing the shape of each part in plan view. Moreover, FIG. 17 is the figure which showed typically the figure corresponding to the arrow view of the XVII-XVII cross section in FIG. 16 and 17, a description will be mainly given of the hollow portion W formed inside the housing 10 including the introduction region 120 on the lid member 100 side and the discharge region 220 on the storage member 200 side. In FIG. 17, the seal member 40 is omitted.

  As shown in FIG. 17, the cavity W is a space that includes an upper introduction region 120 and a lower discharge region 220 with the filter 30 interposed therebetween. The hollow portion W is formed by fixing the first protrusion 125 of the lid member 100 and the second protrusion 225 of the storage member 200 with the filter 30 sandwiched between the outer periphery of the filter region 33. . The top surface 310 of the cavity portion W is formed by a surface 120C (see also FIG. 3) that constitutes the introduction region 120 of the lid member 100 and that is provided with the connection holes 109 and 113. Further, the bottom surface 320 of the cavity W is formed by a surface 220 </ b> C that constitutes the discharge region 220 and is inclined toward the connection hole 209. Further, the inner wall surface 330 of the cavity W is formed by the first protrusion 125 and the second protrusion 225.

  In the cell capture device 1 having such a configuration, the flow of the liquid when the test liquid or the treatment liquid is flowed is indicated by arrows in FIGS. 16 and 17. The liquid (test solution or treatment liquid) introduced into the cavity W (introduction region 120) from the perforation 109 or 113 passes through the through-hole provided in the filter region 33 of the filter 30 and is discharged into the discharge region 220. It is thought that it moves toward the side perforations 209.

  At this time, it is conceivable that the liquid (the test liquid or the processing liquid) moves along a shorter path toward the perforation 209 connected to the discharge channel. Therefore, when viewed from above, as shown in FIG. 16, it is estimated that the hole proceeds as straight as possible from the hole 109 or 113 to the hole 209. When viewed from the side, as shown in FIG. 17, the liquid moves through the introduction region 120 to the filter region 33 where the through hole of the filter 30 is formed, and passes through the through hole when reaching the filter region 33. Thus, it is estimated that the gas flows to the lower discharge area 220 side. Therefore, a path that is assumed to be a liquid flow when there is no obstacle in the flow of liquid from the perforation 109 or 113 to the perforation 209 is defined as a virtual liquid flow line S as shown in FIGS. 17 shows.

  Here, when a liquid (test liquid or processing liquid) is introduced into the cavity W, the liquid flow is relatively large in the region close to the virtual liquid flow line S, and it is assumed that bubbles are contained in the cell trapping device 1. Even if it is mixed, the perforation 109 or the perforation 113 on the upper introduction flow path side is close, so that it is considered that discharge to the outside is easy. On the other hand, in the region away from the virtual liquid flow line S, the flow of liquid is small so that it takes time for the liquid to reach the region from the perforation 109 or the perforation 113 in the first place, and the perforation on the upper introduction flow channel side. 109 or the perforation 113 is also away from the liquid, so that the liquid tends to stay. Furthermore, since the inner wall surface 330 of the cavity W formed by the first protrusion 125 and the second protrusion 225 is provided further outward than the peripheral edge of the filter region 33, However, the retention of the liquid is remarkable on the outside. In addition, when bubbles are mixed in the cell trapping device 1, there is a possibility that the bubbles will stay in the vicinity of the inner wall surface 330 outside the filter region 33. If bubbles remain in such a region and the bubbles become large, the bubbles expand onto the filter region 33, making it difficult to sufficiently capture cells. Moreover, since it is difficult to remove bubbles, the accuracy of measurement when confirming cells after being trapped in the filter region 33 may be reduced. As described above, in the region away from the virtual liquid flow line S, there is a high possibility that the bubbles will stay in the vicinity of the filter 30. If the bubbles stay, it may affect the capture and observation of the cells. In the cell trapping device 1 according to the above-described embodiment, the region away from the virtual liquid flow line S in which bubbles are likely to stay and increase in size is the corner 127 (227) shown in FIG. In the vicinity of the corner 127 (227), the outer periphery of the filter region 33 is also away from the virtual liquid flow line S, and the speed at which the liquid flows becomes slow. The outer periphery of the filter region is obtained by connecting the ends of the outermost through holes in the filter region.

  Therefore, in the cell trapping device 1, the perforations 109 and 113 that are the ends of the introduction flow channels 101 and 102 on the cavity W side and the perforations 209 that are the ends of the discharge channel 201 on the cavity W side are connected. Based on the distance between the virtual liquid flow line S indicating the liquid flow and a predetermined point on the outer periphery of the filter region 33, the distance between the inner wall surface 330 of the cavity W and the predetermined point is changed. ing. Specifically, in the vicinity of a corner 127 (227) where the distance between a predetermined point on the outer periphery of the filter region 33 and the virtual liquid flow line S is larger than the other corners, the first protrusion 125 and the first The shape of the two protruding portions 225 is changed to have a shape having a chamfered portion with chamfered corners 127 (227). Thereby, in the area where chamfering has been performed, the distance between the inner wall surface 330 of the cavity W and the filter area 33 at a predetermined point on the outer periphery is shorter than before the change. Thereby, since it is possible to suppress the bubbles from staying in the vicinity of the region where the chamfer is provided, in the vicinity of the filter 30, it is possible to prevent the bubbles from becoming large. Here, the distance from the inner wall surface 330 at a predetermined point on the outer periphery of the filter region 33 and the distance from the virtual liquid flow line S at the predetermined point on the outer periphery of the filter region 33 both indicate the shortest distance. .

  In the cell trapping device 1, the top surface 310 of the internal cavity including the introduction region 120 and the side surface of the first protrusion 125 that constitutes the inner wall surface 330 of the cavity W (of the lid member 100 side). And are connected by a curved surface. Thereby, even if the bubbles flow into the region outside the outer periphery of the filter region 33, the bubbles are prevented from staying in the region connecting the top surface 310 and the inner wall surface 330, and the bubbles are discharged from the cavity W. Can be promoted.

  Here, an example of changing the shape of the inner wall surface of the cavity outside the filter region will be described with reference to FIG. 18A and 18B both show the shape of the cavity W and the filter region 83 in plan view. In addition, two end flow channel end portions 81 attached to the upper side of the filter and a discharge flow channel end portion 82 attached to the lower side of the filter are shown together. A virtual liquid flow line S up to the end 82 of the discharge channel is shown.

  First, as shown in FIG. 18 (A), the filter region 83 has a hexagonal shape instead of a square, and two end portions 81 of the introduction flow path are attached to two opposite sides across the hexagonal center, and the discharge flow path Is attached near the center of the filter region 83. This configuration corresponds to the case where the cavity W inside the housing and the filter region are changed to a hexagonal shape with respect to the cell trapping device 1 shown in the above embodiment. In this case, the distance from the virtual liquid flow line S at a predetermined point on the outer periphery of the filter region 83 is increased in the vicinity of the region separated from the region where the end 81 of the introduction flow path is attached. Therefore, by forming the chamfered portions P1 and P2 in the corresponding region of the inner wall surface of the cavity W, it is possible to prevent bubbles from staying in this region. In FIG. 18A, not only the chamfered portion P1 is formed in a region where the distance from the virtual liquid flow line S at a predetermined point on the outer periphery of the filter region 83 is the largest, but P1 is formed. The chamfered portion P2 is also formed in a region where the distance from the virtual liquid flow line S is smaller than that in the region. Further, in the inner wall surface in the cavity W which is rearward with respect to the direction from the end portion 81 of the two introduction flow paths to the end portion 82 of the discharge flow path, and in a region where the distance from the end portion 81 is large However, since the flow rate is reduced and bubbles are likely to stay, the chamfered portion P3 can be formed. Thus, based on the distance from the virtual liquid flow line S at a predetermined point on the outer periphery of the filter region 83, the cavity portion W is changed so that the distance from the inner wall surface of the cavity portion W at the predetermined point changes. The shape of the inner wall surface can be changed as appropriate.

  Next, FIG. 18B shows an example in which the shape of the chamfered portion P of the cavity W is changed as compared with the cell trapping device 1 shown in FIG. In FIG. 18B, the arrangement of the filter region 83, the end 81 of the introduction channel, and the end 82 of the discharge channel is the same as that of the cell trapping device 1 shown in FIG. On the other hand, the shape of the chamfered portion P is not a shape with the corner 127 (227) cut off as in the cell trapping device 1, but a curved surface. In this way, when the chamfered portion is formed in a curved shape, it is possible to suppress the bubbles from staying at the corners between the cut-off surface and the surface adjacent to the surface, thereby further preventing the bubbles from staying. it can. In addition, the modification which makes a chamfer part curved surface is applicable also to a cell capture device provided with structures other than the cell capture device 1 demonstrated in the said embodiment.

  As mentioned above, although embodiment of this invention was described, this invention is not limited to the said embodiment, A various change can be made. For example, in the above-described embodiment, the shape of the lid member and the storage member of the cell trapping device is a substantially square shape, but it may be a pentagon, a hexagon, or other polygons, and may be a circle or an ellipse.

  In the above embodiment, the correspondence between the fixing member and the guide portion is 1: 1, but a plurality of fixing members may be attached to one guide portion. Further, the shape of the fixing member is not limited to the above embodiment and can be changed as appropriate.

  Furthermore, the method of fixing the lid member, the storage member, and the filter in the cell trapping device including the fixing member is not limited to the above embodiment, and other modes can be adopted. As another aspect, for example, there is a method of fixing the lid member and the storage member by an adhesive or welding after sandwiching the filter member.

DESCRIPTION OF SYMBOLS 1 ... Cell capture device, 1A ... Cell capture system, 10 ... Housing, 30 ... Filter, 36 ... Positioning hole, 40 ... Sealing member (gasket), 60 ... Fixing member, 100 ... Cover member, 101, 102 ... Introduction Flow path, 120 ... introduction area, 125 ... first protrusion, 130 ... first fitting part, 151 ... lid member side guide part, 200 ... housing member, 201 ... discharge flow path, 220 ... discharge area, 225 ... 2nd protrusion part, 230 ... 2nd fitting part, 240 ... Bottomed hole (hole part), 251 ... Storage member side guide part.

Claims (6)

A housing having a lid member having an introduction flow path for introducing the test liquid into the interior, and a storage member having a discharge flow path for discharging the test liquid to the outside;
Between the introduction flow path and the discharge flow path, a through-hole is provided inside a cavity having an inner wall surface formed by the lid member and the storage member, and the test liquid passes therethrough. A filter having a filter region formed in the thickness direction on the inner side and supported by the lid member and the storage member on the outer side of the filter region of the filter;
A cell capture device comprising:
The cell trapping device in which the shape of the inner wall surface and the shape of the outer periphery of the filter region in plan view are different from each other.   In plan view, the distance from the inner wall surface at a predetermined point on the outer periphery of the filter region is the predetermined point, the end of the introduction channel on the cavity side, and the cavity of the discharge channel. The cell trapping device according to claim 1, wherein the cell trapping device is changed based on a distance from a virtual liquid flow line indicating a flow of the test liquid connecting between the end portions on the side. The filter region has a substantially rectangular shape,
The end of the introduction channel on the side of the cavity is provided at a position corresponding to two opposing corners of the four corners of the filter region in the lid member,
The end of the discharge channel on the cavity portion side is provided at the center of the region corresponding to the filter region on the bottom surface of the cavity of the storage member,
The inner wall surface of the hollow portion in plan view has a substantially rectangular shape corresponding to the filter region, and two of the four corners of the filter region are not provided with the end portion on the hollow portion side of the introduction channel. The cell capturing device according to claim 1, wherein the cell capturing device has a chamfered portion in a region corresponding to a corner.   The cell capture device according to claim 3, wherein the chamfered portion is formed in a curved surface shape.   The cell trapping device according to any one of claims 1 to 4, wherein the hollow portion is a curved surface from the top surface on the lid member side to the inner wall surface. The cell capture device according to any one of claims 1 to 5,
A test liquid supply means for supplying a test liquid to the introduction channel of the cell capture device;
A processing liquid supply means for supplying a processing liquid for processing cells captured by the filter by passing through the filter with respect to the introduction channel of the cell capturing device;
A cell capture system comprising: a selection unit that selects a liquid to be supplied to the cell capture device from the test liquid and the treatment liquid.

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