JP2015198595A - Cell-capturing device and cell-capturing apparatus - Google Patents

Abstract

PROBLEM TO BE SOLVED: To make is possible to suppress mixture of air bubbles to the inside, and efficiently perform processing regarding cell capture.SOLUTION: In a cell-capturing device, seal parts 305, 405, 905 of flow channel connecting parts 30, 40, 90 are provided for a flow channel connecting the inside and the outside of the cell-capturing device 1, and has a structure of switching opening and closing of the flow channel by the presence or absence of connection with an external flow channel, so that mixture of outside air into the cell-capturing device 1 can be prevented, and generation of air bubbles inside the cell-capturing device 1 when circulating processing liquid or test solution in the device can be suppressed. Further, reduction in efficiency of cell capture can be suppressed by suppressing generation of air bubbles, and processing regarding cell capture can be efficiently performed.

Description

  The present invention relates to a cell capturing device and a cell capturing apparatus.

  Cancers, also called “malignant tumors”, have seriously hindered the life support of living organisms when they progress, and various treatment methods have been studied. There is the presence or absence of cancer metastasis as a guideline when deciding the treatment policy for cancer patients. Metastasis is caused by cancer cells invading blood vessels and lymphatic vessels, spreading through them through the body, and moving to other organs. Therefore, the measurement of the presence and amount of cancer cells in the blood is important information for predicting cancer metastasis. Cancer cells that circulate in the human body through such blood vessels or lymph vessels are called circulating tumor cells (CTCs).

  As a conventional technique for capturing CTC for examination of metastatic cancer or evaluation of an anticancer agent, for example, as shown in Patent Document 1, a method of capturing CTC with a filter is known. Patent Document 1 discloses a manufacturing method using a filter semiconductor technology, the shape of a cell unit housing a filter, and the structure of a flow channel for flowing blood and processing liquid. Specifically, a configuration is shown in which cancer cells are captured by flowing blood containing cancer cells into a cell unit containing a filter, and cells existing on the filter are identified as cancer cells by staining. Yes. In Patent Documents 2 and 3, a cell dispersion containing at least one of cancer cells and immune cells is allowed to pass through, and at least one of the cancer cells and the immune cells is subjected to physical action, chemical action and A body fluid treatment system including a cell treatment cartridge capable of imparting at least one kind of physiologically active action is disclosed. Patent Document 4 discloses a microfluidic device including a polydimethylsiloxane (PDMS) upper member having a sample supply port above and below a nickel substrate having fine through holes in a nickel substrate and a lower member having a sample discharge port. Has been.

US Patent Application Publication No. 2011/0053152 JP 2010-227011 A JP 2010-75191 A JP 2011-163830 A

  However, when the apparatuses described in Patent Documents 1 to 4 are used, bubbles may be mixed in when performing an operation of capturing cancer cells in blood. In particular, when air bubbles are mixed in the flow path near the filter, the air bubbles impede the flow of the liquid.For example, the blood is not sufficiently supplied to the entire surface of the filter, so that the capture efficiency is greatly reduced. There is a concern that the efficiency of sample processing will be significantly reduced.

  The present invention has been made in view of the above, and a cell trapping device capable of efficiently performing a process related to trapping cells by suppressing the mixing of bubbles therein, and a cell trapping apparatus including the cell trapping device It is an issue to provide.

  In order to achieve the above object, a cell capturing device according to the present invention includes an introduction flow path for introducing a test liquid or a processing liquid for processing cells in the test liquid, and the introduced flow path A discharge flow path for discharging the test liquid or the processing liquid to the outside and a plurality of through holes are formed in the thickness direction, and the test flow is formed on the flow path between the introduction flow path and the discharge flow path. A cell capture device comprising: a filter disposed so that the liquid or the treatment liquid passes through the through-hole; and a housing in which the introduction flow path, the discharge flow path, and the filter are provided. The introduction flow path and the discharge flow path can each be connected to an external flow path for allowing the test liquid or the processing liquid to flow between the outside and the outside on the surface of the housing. Each of the flow path connecting portions is the test liquid or the treatment, respectively. An open / close valve that opens and closes the flow of the liquid, and the open / close valve opens the flow of the test liquid or the processing liquid when the external flow path is connected to the flow path connection portion. And when the said external flow path is not connected with respect to the said flow-path connection part, the distribution | circulation of the said test liquid or the said process liquid is closed.

  According to the above-described cell trapping device, when the external channel is connected by the open / close valve of the channel connecting portion, the flow of the test liquid or the processing solution in the cell trapping device is opened, while the external channel Is not connected, the flow of the test liquid or the processing liquid is closed. That is, by having a configuration that switches between opening and closing depending on whether or not an external flow path is connected, it is possible to prevent outside air from being mixed into the inside of the cell trapping device, and the treatment liquid is contained in the cell trapping device. Alternatively, it is possible to suppress the generation of bubbles inside when the test solution is circulated. Further, by suppressing the generation of bubbles, it is possible to suppress a decrease in the efficiency of cell trapping, and it is possible to efficiently perform a process related to cell trapping.

  Here, as a configuration that effectively exhibits the above-described operation, for example, the on-off valve is configured by an elastic member that is provided so as to seal the introduction channel or the discharge channel in the channel connection portion. An embodiment is mentioned.

  Further, as another configuration that effectively exhibits the above-described operation, for example, the on-off valve is a sealing on a flat plate provided to seal the introduction flow path or the discharge flow path at the flow path connection portion The aspect comprised by a member and the elastic member which supports the said sealing member so that a movement is possible is mentioned.

  Further, in the above-described cell trapping device, it is preferable that the flow path connecting portions of the introduction flow path and the discharge flow path are provided on the same plane of the casing. As described above, by providing the flow path connection portion on the same plane of the housing, it is possible to increase the work efficiency related to connection and disconnection with the external flow path.

  The cell capture device according to the present invention comprises the above-described cell capture device, a test liquid storage container that stores the test liquid, the test liquid storage container, and the introduction flow path of the cell capture device. An external flow channel to be connected, and a test solution flow channel having a device connection portion connectable to the flow channel connection portion of the introduction flow channel, and captured by the filter by passing through the filter A processing liquid storage container for storing the processing liquid for processing cells, an external flow path for connecting the processing liquid storage container and the introduction flow path of the cell trapping device, A treatment liquid flow path having a device connection portion connectable to the flow path connection portion, and an external flow path connected to the discharge flow path of the cell trapping device, wherein the flow path connection of the discharge flow path Device connection that can be connected to A flow path for discharging outside the system, a selection means for selecting a liquid to be supplied to the cell trapping device from the test liquid and the treatment liquid, and the storage of the test liquid based on a selection result of the selection means Liquid feeding means for supplying a test liquid from a container or a treatment liquid from the treatment liquid storage container to the cell capture device.

  According to the above-described cell trapping device, when the test liquid channel, the processing liquid channel, and the external discharge channel, which are external channels, are connected by the open / close valve of the channel connection part in the cell trapping device. Opens the flow of the test liquid or the processing liquid in the cell trapping device, while the test liquid flow path, the processing liquid flow path, and the outside discharge flow path are not connected. Liquid distribution is closed. That is, by having a configuration that switches between opening and closing depending on whether or not an external flow path is connected, it is possible to prevent outside air from being mixed into the inside of the cell trapping device, and the treatment liquid is contained in the cell trapping device. Alternatively, it is possible to suppress the generation of bubbles inside when the test solution is circulated. Further, by suppressing the generation of bubbles, it is possible to suppress a decrease in the efficiency of cell trapping, and it is possible to efficiently perform a process related to cell trapping.

  In addition, it is preferable that the above-described cell capturing device further includes a first moving unit that movably supports the cell capturing device.

  By providing the first moving means for movably supporting the cell trapping device in this way, for example, in a state where the test liquid flow path, the processing liquid flow path, and the external discharge flow path that are external flow paths are fixed. By connecting the cell capturing device with the first moving means, the cell device and the external channel can be easily connected and disconnected. Therefore, it is possible to suppress the generation of bubbles when the processing solution or the test solution is circulated in the cell trapping device, and it is possible to increase the work efficiency related to connection and disconnection with the external flow path.

  Also, supported by the second moving means, one end side can be connected to the device connection part of the processing liquid flow path, and the other end side can be connected to the device connection part of the extra-system discharge flow path. It is preferable to further include a bypass flow path.

  By further providing the bypass flow path in this way, when bubbles are included in the treatment liquid flow path from the treatment liquid storage container, the bypass flow path is formed before connecting the cell capture device to the treatment liquid flow path. Air bubbles can be removed from the treatment liquid flow path by flowing the treatment liquid into the treatment liquid flow path while being connected to the treatment liquid flow path and the out-of-system discharge flow path. Generation of bubbles when the liquid or the test liquid is circulated can be more effectively suppressed.

  ADVANTAGE OF THE INVENTION According to this invention, the cell capture device which suppresses mixing of the bubble to an inside and can perform the process which concerns on cell capture efficiently, and the cell capture apparatus containing this cell capture device are provided.

It is a figure explaining schematic structure of the cell capture device concerning the embodiment of the present invention. It is a figure explaining schematic structure of the cell capture device which comprises a cell capture device. It is a figure explaining schematic structure of the cell capture device which comprises a cell capture device, and is the figure which added the flow path in a cell capture device with the broken line with respect to sectional drawing along the III-III line of FIG. It is a figure explaining the schematic structure of the flow-path connection part of a cell capture device. It is a figure explaining the connection method at the time of connecting an external flow path with respect to a cell capture device. It is a figure explaining schematic structure at the time of connecting an external channel to a cell capture device. It is a figure explaining the structure at the time of connecting an external flow path to the flow-path connection part of a cell capture device. It is a figure explaining the modification about the flow-path connection part of a cell capture device, and the modification of the device connection part of a flow path. It is a figure explaining the modification about the flow-path connection part of a cell capture device, and the modification of the device connection part of a flow path. It is a figure explaining the modification about the flow-path connection part of a cell capture device, and the modification of the device connection part of a flow path. It is a figure explaining the modification of a cell trapping device. It is a figure explaining the modification of a cell trapping device.

  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 diagram illustrating a configuration of a cell trapping apparatus according to this embodiment. The cell trapping device is a device that traps cells contained in a test liquid by filtering a cell dispersion liquid serving as a test liquid with a filter. In addition, the cells captured by the filter are stained using a treatment liquid such as a staining liquid, thereby identifying the cells and counting the number of individual cells. An example of the cell dispersion used as a test solution is blood. In addition, the cell capture device, for example, allows red blood cells, platelets and white blood cells (hereinafter collectively referred to as “blood cell components”) contained in blood to pass through from blood containing circulating cancer cells called CTC. Therefore, it is preferably used for the purpose of capturing CTC.

  As shown in FIG. 1, a cell capture device 100 includes a cell capture device 1 in which a filter for capturing cells is provided, and a soft tube for supplying a treatment liquid (reagent) to the cell capture device 1. And a flow path 4 (test liquid flow path) made of a soft tube for supplying a test liquid to the cell capture device 1. In FIG. 1, the cell capturing device 1 and the flow path 3 and the cell capturing device 1 and the flow path 4 are separated from each other. However, when the cell capturing apparatus 100 is used to perform processing for capturing and observing cells. These are connected to. The same applies to the flow path 9 described later.

  A plurality of processing liquid storage containers 5 in which different processing liquids (reagents) are placed are provided on the upstream side of the flow path 3. The treatment liquid to be introduced into the treatment liquid storage container 5 includes a staining liquid for staining cells, a washing liquid for washing cells captured by a filter, a fixing liquid for protecting cells from spoilage, and the like. Examples include permeate for allowing the liquid to penetrate into the cells. The plurality of treatment liquid storage containers 5 shown in FIG. 1 are sealed by the cover member 5A, but this configuration is not particularly limited.

  A soft tube 6 is inserted into each of the plurality of processing liquid storage containers 5 to form individual flow paths (processing liquid flow paths). These flow paths are connected to the selection valve 8, and the processing liquid to be connected to the flow path 3 is selected by rotating the selection valve 8, and the processing in which the selected processing liquid is accommodated. The soft tube 6 inserted into the liquid storage container 5 and the flow path 3 are connected.

  A test liquid storage container 10 in which blood containing cells is stored as a test liquid is connected to the flow path 4 connected to the cell trapping device 1 during use. The cell trapping device 1 is configured to supply either one of the treatment liquid and the test liquid at the same time, rather than simultaneously. Control of which of the processing liquid and the test liquid is supplied is switched by valves 12 and 13 attached to the flow paths 3 and 4 respectively. For example, when supplying the test solution to the cell capture device 1, the valve 12 is closed and the valve 13 is opened. As the valves 12 and 13, pinch valves that pressurize and deform the soft tube to block the flow can be used.

  In the drawings after FIG. 1, the XYZ axes are shown for explanation. Among these, the XY plane indicated by the XY axis corresponds to a so-called horizontal plane, and the Z axis indicates a vertical line. Moreover, it is not for the purpose of showing the direction of the arrangement of the cell trapping apparatus 100 according to the present embodiment as a whole, but for illustrating the configuration of the cell trapping device 1. That is, the treatment liquid storage container 5 and the test liquid storage container 10 constituting the cell capturing device 100 are containers for storing liquids, respectively, and the bottom surface is arranged in a direction intersecting the Z axis, On the other hand, an opening of the container is provided above the Z axis.

  When supplying any one of the treatment liquid and the test liquid to the cell capture device 1, the flow paths 3 and 4 on the upstream side of the cell capture device 1 are connected to the cell capture device 1, A flow path 9 (external discharge flow path) made of a soft tube on the downstream side of the cell capture device 1 is connected to the cell capture device 1. Then, the liquid is supplied by sucking the target liquid by driving a pump 14 (supply means) provided on the flow path 9. The pump 14 has a structure capable of changing the flow rate of the liquid in the flow path by changing the rotation speed. As the pump 14, for example, a peristaltic pump (peristaltic pump) that sequentially moves a peristaltic point by pressurizing the soft tube can be used. By driving the pump 14, liquids such as a processing solution and a test solution flow in the direction toward the cell trapping device 1 through the flow path 3 or the flow path 4 and are supplied to the cell trapping device 1. The liquid that has passed through the cell trapping device 1 flows into the waste liquid container 16 via the flow path 9. With these structures, the cells in the test solution are captured by a filter provided on the flow path in the cell capturing device 1, and the cells are stained with the staining solution.

  The end portions of the flow paths 3, 4, 9 on the cell capture device 1 side are provided with device connection portions 31, 41, 91 that are hollow like injection needles and whose tip portions are processed into a needle shape, respectively. The device connection portions 31, 41, 91 are inserted into the cell capture device 1, whereby the flow paths 3, 4, 9 and the cell capture device 1 are connected. This configuration will be described later. The device connection parts 31, 41, 91 are preferably fixed by a support or the like. When the cell capture device 1 is moved in a state where the device connection portions 31, 41, 91 are supported, and the cell capture device 1 is attached and detached, the device connection portions 31, 41, 91 are individually moved. In comparison, the working efficiency is improved.

  The cell trapping device 1 is movably supported by a moving device 17 (first moving means). The moving device 17 includes a fixed base 18, a mounting base 19 that can move along the XY plane with respect to the fixed base, and a driving motor 20. The mounting base 19 is rotated by the rotation of the driving motor 20. It moves in the X-axis direction or the Y-axis direction (the moving device 17 of this embodiment moves in the Y-axis direction). The cell trapping device 1 can be moved along with the movement of the mount 19 by being fixed above the mount 19. Although the moving direction is not limited to the above, when the cell capturing device 1 is moved by the moving device 17, connection / disconnection between the channel 3, 4, 9 and the cell capturing device 1 is switched.

  The control of each of the above parts is performed by control by a control part (selection means, liquid feeding means / not shown). Specifically, the selection valve 8, the valves 12 and 13 are driven, and the cell capture device 1 is moved by the moving device 17 according to instructions from the control unit. The control unit is provided with a program input function for inputting a program enabling control of driving, stopping, etc. for each of the above-described units, and each device is sequentially switched as described above according to the program input thereby. A drive mechanism for operation is added. The control unit 30 selects a line through which the liquid flows, and based on the selection result, the control unit instructs each unit to open / close the valve and drive the pump.

(Cell capture device)
Next, the cell trapping device 1 will be described with reference to FIGS. FIG. 2 is a top view of the cell trapping device 1, and FIG. 3 is a diagram in which a flow path in the cell trapping device 1 is added with a broken line with respect to a cross-sectional view taken along line III-III in FIG. .

  The cell trapping device 1 is configured such that a filter 57 having a plurality of through holes 61 is sandwiched between a lid member 58 and a storage member 59. The filter 57 is disposed in a space formed inside the lid member 58 and the storage member 59 when they are combined. The filter 57 is made of, for example, metal and has a plurality of through holes 61 formed in the thickness direction. Further, the lid member 58 and the storage member 59 constitute a casing of the cell trapping device 1.

  The lid member 58 of the cell trapping device 1 includes a flow path 3A (introduction flow path) connected to the flow path 3 formed of a soft tube, and a flow path 4A (introduction flow path) connected to the flow path 4. And an introduction region 62 which is formed above the filter 57 in communication with the flow paths 3A and 4A and serves as a space for guiding the liquid to the through-hole 61 of the filter 57. That is, in the present embodiment, the “introduction channel” refers to the channels 3 </ b> A and 4 </ b> A provided inside the cell trapping device 1 among the channels. Further, the “test solution channel” and the “treatment solution channel” refer to the channels 3 and 4 connected to the upstream sides of the channels 3A and 4A of the cell trapping device 1, respectively.

  The storage member 59 of the cell trapping device 1 is formed below the filter 57 so that the central portion is deeper than the peripheral portion, and discharges the liquid that has passed through the through hole 61 of the filter 57 to the outside. A discharge area 63 serving as a space is provided. Further, the storage member 59 is provided with a flow path 9A (discharge flow path) that communicates with the discharge area 63 and is connected to the flow path 9 and discharges the liquid in the discharge area 63 to the outside. . The through-hole 61 provided in the filter 57 sandwiched between the lid member 58 and the storage member 59 is set to a size that prevents the cells to be captured from passing through.

  Further, in the flow path 3A in the cell trapping device 1, the surface of the housing is on the end opposite to the end on the filter 57 side for circulating the processing liquid between the outside. A flow path connection portion 30 to which the flow path 3 as an external flow path can be connected is provided. Similarly, in the flow path 4A, on the surface of the housing and on the end opposite to the end on the filter 57 side is an external flow path for allowing the test liquid to flow between the outside. A flow path connecting portion 40 to which the flow path 4 can be connected is provided. Further, in the flow path 9A, on the surface of the housing, on the end opposite to the end on the filter 57 side, the test solution and the processing solution that have passed through the cell trapping device 1 are placed between the outside. A flow path connecting portion 90 is provided to which a flow path 9 that is an external flow path for circulating the air can be connected. As shown in FIG. 2, the flow path connection portions 30, 40, and 90 are provided on the same plane of a casing that includes a lid member 58 and a storage member 59. Thereby, the flow paths 3, 4, and 9 can be connected to the cell trapping device 1 at a time, and the working efficiency can be improved.

  The structure of the flow path connecting portions 30, 40, 90 will be described with reference to FIG. In FIG. 4, the flow path connection portion 30 will be described, but the flow path connection portions 40 and 90 also have the same configuration. The flow path connecting portion 30 is connected to the flow path 3A and is provided on the surface of the housing with the connection flow path 301 having a larger inner diameter than the flow path 3A, and is connected to the flow path 3A among the ends of the connection flow path 301. And an opening 302 that is connected to the connection channel 301 at the end opposite to the end side to be connected and has an inner diameter larger than that of the connection channel 301. Further, the opening 302 is provided with a seal portion 305 made of an elastic member provided to seal the flow path. By sealing the flow path with the seal portion 305, it is possible to prevent the outside air from flowing into the cell trapping device 1 before use. The seal portion 305 is made of elastic rubber or the like, and functions as an on-off valve that opens and closes the flow of the processing liquid (or test liquid). That is, when the device connection part 31 is inserted into the connection flow path 301 and the flow path 3 and the cell capture device 1 are connected, the seal part 305 is treated with the treatment liquid or the test through the device connection part 31. When the liquid flow is released and the device connection unit 31 is detached from the cell capture device 1 and the connection between the flow channel 3 and the cell capture device 1 is disconnected, the seal unit 305 seals the flow channel again. It has a function.

(Connection method of cell capture device)
Next, a method for connecting the flow paths 3, 4, 9 to the flow path connection portions 30, 40, 90 will be described with reference to FIGS. 5 to 7. In FIG. 5, description will be made using an example of the flow path 3. First, when connecting the flow path 3 to the cell trapping device 1, the device connection portion 31 of the flow path 3 faces the seal portion 305 provided in the opening 302 of the flow path connection portion 30, and The connection channel 301 is disposed along the extending direction. In the cell trapping device 1 according to this embodiment, since the connection channel 301 extends in the Y-axis direction, the needle-like device connection portion 31 is also arranged so as to be coaxial along the Y-axis direction. Similarly, the device connection part 41 of the flow path 4 and the device connection part 91 of the flow path 9 are also arranged at positions corresponding to the flow path connection parts 40 and 90, respectively. Thereafter, by moving the cell capture device 1 in the Y-axis direction by the moving device 17, the device connection portion 31 is inserted into the connection flow path 301 of the cell capture device 1 as shown in FIG. Similarly, the device connection unit 41 is inserted into the connection channel 401 of the cell capture device 1, and the device connection unit 91 is inserted into the connection channel 901 of the cell capture device 1.

  At this time, in the cell trapping device 1 according to the present embodiment, as shown in FIG. 7A, before the device connection portion 31 is inserted, the connection flow path 301 is connected to the flow path 3A by the seal portion 305. While sealing the flow of the processing liquid (or test liquid), as shown in FIG. 7B, the connection channel 301 of the channel connection unit 30 is connected to the connection channel 301 so that the device connection unit 31 breaks through the seal unit 305. When inserted, the seal portion 305 is opened only in a region corresponding to the outer diameter of the device connection portion 31. The seal portion 305 is a member having elasticity, and since an opening larger than the outer diameter of the device connection portion 31 is suppressed from being formed in the seal portion 305, the cell is removed from the gap between the seal portion 305 and the device connection portion 31. It is possible to prevent air from entering the trapping device 1 and to prevent the liquid in the cell trapping device 1 from leaking to the outside through the gap between the seal portion 305 and the device connection portion 31. Furthermore, when the device connection part 31 is removed from the connection flow path 301, the seal part 305 is configured to close the flow path 3A as before the device connection part 31 is inserted. As described above, the seal portions 305, 405, and 905 are provided in the flow path that connects the inside and the outside of the cell trapping device 1, and have a configuration that switches between opening and closing depending on whether or not the external flow path is connected. Thus, it is possible to prevent outside air from being mixed into the inside of the cell trapping device 1 and to suppress the generation of bubbles when the processing solution or the test solution is circulated in the cell trapping device 1. can do. Further, by suppressing the generation of bubbles, it is possible to suppress a decrease in the efficiency of cell trapping, and it is possible to efficiently perform a process related to cell trapping.

  Further, in the cell trapping device 1 according to the present embodiment, the flow path connecting portions 30, 40, 90 are formed on the same plane of the housing, and the connection flow paths 301, 401, 901 are in the same direction (Y-axis direction). ), The attachment of the device connection part 31 of the flow path 3, the device connection part 41 of the flow path 4 and the device connection part 91 of the flow path 9 to the cell capture device 1 is one of the cell capture devices 1. It can be achieved only by moving in the direction. Specifically, the device connection parts 31, 41, 91 are juxtaposed in correspondence with the arrangement of the connection flow paths 301, 401, 901, and the device connection parts 31, 41, 91 are connected to the connection flow paths 301, 401, respectively. , 901 can be attached at a time by moving the cell capture device 1 in the Y-axis direction so as to be inserted into 901. For this reason, connection with the cell capture device 1 and the three flow paths 3, 4, 9 can be performed by a simpler operation, and an improvement in work efficiency can be achieved.

(Modification)
Next, modified examples of the cell trapping device and the cell trapping apparatus according to the present invention will be described. First, after describing a modified example of the flow path connecting portion of the cell trapping device, a modified example of the cell trapping device will be described.

(Modification 1)
FIG. 8 is a view for explaining a first modification of the flow path connection part 30 of the cell trapping device. FIG. 8A shows the flow path connection portion before connecting the flow paths, and FIG. 8B shows the flow path connection portion in a state where the flow paths are connected. The flow path connection portion 30A of the cell trapping device shown in FIG. 8 differs from the flow path connection portion 30 described in the above embodiment in the shape of the casing 72 around the opening 302 and the shape of the seal portion. That is, in the flow path connecting portion 30A shown in FIG. 8, a convex portion 721 having a substantially cylindrical shape around the opening 302 and the lower side of the connection flow channel 301 is formed in the casing 72. A neck portion 722 having an inner diameter narrower than the upper portion is formed below 721 (on the surface side of the casing 72). The seal portion 306 has a shape that covers the upper side of the opening 302 and further covers the upper surface of the convex portion 721 (the surface on which the opening 302 is formed), and a thick coupling portion 306A is formed on the outer peripheral side of the seal portion 306. The sealing portion 306 is fixed in a state in which the end portion (opening 302) of the flow path 3A is closed by fitting the coupling portion 306A to the neck portion 722.

  When such a flow path connection portion 30A is provided, as shown in FIG. 8B, the device connection portion 31 is inserted into the connection flow path 301 of the flow path connection portion 30 so as to break through the seal portion 306. When this is done, the seal portion 306 is opened only in a region corresponding to the outer diameter of the device connection portion 31, and air may be mixed into the cell trapping device 1 from the gap between the seal portion 306 and the device connection portion 31. In addition to being suppressed, the liquid in the cell trapping device 1 is also prevented from leaking outside through the gap between the seal portion 306 and the device connection portion 31. Furthermore, when the device connection part 31 is removed from the connection flow path 301, the seal part 306 is configured to block the flow path 3A as before the device connection part 31 is inserted. Airtightness is maintained.

(Modification-2)
FIG. 9 is a diagram for explaining a second modification of the flow path connection part 30 of the cell trapping device. FIG. 9A shows the flow path connecting portion before connecting the flow paths, and FIG. 9B shows the flow path connecting portion in a state where the flow paths are connected. The flow path connection portion 30B of the cell trapping device shown in FIG. 9 is different from the flow path connection portion 30A described in the above embodiment in that the seal portion 306 functions as an open / close valve that opens and closes the flow path. The difference is that an on-off valve is constituted by a sealing member on a flat plate provided so as to seal the passage 3A and an elastic member that supports the sealing member so as to be movable. Moreover, the shape of the device connection part of the flow path 3 is also changed corresponding to the shape of the flow path connection part 30B.

  Specifically, as shown in FIG. 9A, the flow path connecting portion 30B is connected to the flow path 3A by a connection flow path 301 and a connection flow path 301 having a larger inner diameter than the flow path 3A. Is provided with an opening 303 and an opening 304 that connect the surface of the housing 72 and the connection channel 301. Further, a flat plate-shaped sealing member 312 supported by an elastic member 314 so as to be movable along the extending direction of the connection channel 301 is provided inside the connection channel 301.

  When the flow path 3 and the cell trapping device 1 are not connected to each other by the elastic member 314 (normal time), the sealing member 312 is connected via the O-ring 313 fixed to the sealing member 312. It is attached so as to come into contact with the inner wall surface on the opening 303 side of 301. The O-ring 313 is provided so as to surround the periphery of the end face of the opening 303 on the side of the connection flow path 301 at the normal time. With such a configuration, the flow path in the cell trapping device 1 is normally closed by the O-ring 313 and the sealing member 312.

  The sealing member 312 is further provided with a pin 311 that protrudes to the outside of the housing 72 through the opening 304. The pin 311 has a shape corresponding to the inner diameter of the opening 304 and can slide inside the opening 304. The pin 311 and the sealing member 312 may be fixed to each other by adhesion or the like, or may be separated from each other.

  On the other hand, the device connection portion 31A of the flow path 3 includes a block 33 that covers the periphery of the flow path 3 at the end of the flow path 3 on the cell capturing device 1 side, and an end face of the flow path 3 on the surface of the block 33. And an O-ring 34 fixed so as to surround. The size of the block 33 is such that when the device connection part 31 is brought into contact with the cell trapping device 1 and the flow channel 3 is connected, the block 33 is arranged at the same time when the flow channel 3 is arranged to face the opening 303. The pin 311 protruding from the opening 304 is sized to be pressed.

  Then, as shown in FIG. 9B, the block 3 is brought into contact with the housing 72, so that the pin 311 slides toward the connection flow path 301, whereby the sealing member 312 is moved to the flow path 3A. Move to the side. Thereby, when the sealing member 312 and the opening 303 are separated from each other, the closing of the flow path by the O-ring 313 and the sealing member 312 is released, and the flow path 3, the opening 303, and the connection flow path 301 communicate with each other. The flow path is opened. Further, when the device connection portion 31A of the flow path 3 is separated from the cell trapping device 1, the pressure on the pin 311 by the block 33 is released, and the elasticity of the elastic member 314 returns the sealing member 312 to the normal position. 9A, the gap between the opening 303 and the connection channel 301 is closed by the sealing member 312 and the O-ring 313, and the channel is closed.

  Thus, even when it is set as the structure provided with the opening-and-closing valve by the elastic member which supports the sealing member and the sealing member movably instead of the structure using the sealing part which has elasticity as an opening-and-closing valve. In addition, it is possible to prevent outside air from being mixed into the inside of the cell trapping device, and to suppress the generation of bubbles when the processing solution or the test solution is circulated in the cell trapping device. it can. Further, by suppressing the generation of bubbles, it is possible to suppress a decrease in the efficiency of cell trapping, and it is possible to efficiently perform a process related to cell trapping.

(Modification-3)
FIG. 10 is a diagram for explaining a third modification of the flow path connection part 30 of the cell trapping device. FIG. 10A shows a flow path connection portion before connecting the flow paths, and FIG. 10B shows the flow path connection portion in a state where the flow paths are connected. The flow path connection part 30C of the cell trapping device shown in FIG. 10 is obtained by changing the configuration of the sealing member and the elastic member that function as an on-off valve, compared to the flow path connection part 30B shown as the above modification. is there.

  The connection flow path 301, the openings 303 and 304, the sealing member 312 and the O-ring 313 fixed to the sealing member 312 in the flow path connection section 30C are the same as the flow path connection section 30B shown in FIG. However, the member that movably supports the sealing member 312 is replaced by a notch spring 316 instead of the elastic member 314, and the notch spring 316 is connected to the sealing member 312 at one end side and the other end. It is fixed to the side wall surface of the connection channel 301 on the side. Further, the pins 315 slidably provided in the opening 304 are movable with respect to each other without being fixed to the sealing member 312.

  In such a flow path connecting portion 30C, as shown in FIG. 10B, the pin 315 slides toward the connection flow path 301 by bringing the block 33 into contact with the housing 72. Then, the sealing member 312 is pushed in. Here, of the sealing member 312, the side opposite to the side fixed mainly by the notch spring 316 is moved to the flow path 3 </ b> A side by pressing with the pin 315. Thereby, when the sealing member 312 and the opening 303 are separated from each other, the closing of the flow path by the O-ring 313 and the sealing member 312 is released, and the flow path 3, the opening 303, and the connection flow path 301 communicate with each other. The flow path is opened. When the device connection portion 31A of the flow path 3 is separated from the cell trapping device 1, the pressure on the pin 315 by the block 33 is released, and the sealing member 312 returns to the normal position by the elasticity of the notch spring 316. 10A, the gap between the opening 303 and the connection channel 301 is closed by the sealing member 312 and the O-ring 313, and the channel is closed.

  As described above, even when the configuration of the on-off valve in the flow path connection portion 30C is changed, it is possible to prevent outside air from being mixed into the inside of the cell trapping device, and the treatment liquid is contained in the cell trapping device. Alternatively, it is possible to suppress the generation of bubbles inside when the test solution is circulated. The configuration of the on-off valve can be variously changed and is not limited to the configuration shown in the above embodiment.

(Modification-4)
Next, a first modification of the configuration of the cell trapping device 100 will be described. The cell capture device 100A shown in FIG. 11 is obtained by changing the following points compared to the cell capture device 100 shown in FIG. In other words, one end side can be connected to the device connection portion 31 of the flow path 3 (treatment liquid flow path), and the other end side can be connected to the device connection section 91 of the flow path 9 (external discharge flow path). The bypass cartridge 70 provided with the flow path 71 is newly provided.

  The bypass cartridge 70 has a bypass channel 71 formed inside a casing 72. When bubbles are contained in the channel 3 from the processing liquid storage container 5, the bypass cartridge 70 is passed through the cell trapping device 1. It is used to eliminate bubbles from the flow path 3 by flowing the processing liquid into the flow path 3 with the bypass cartridge 70 connected before connecting to the flow path 3.

  At both ends of the bypass flow channel 71 in the bypass cartridge 70, that is, at portions connected to the device connection portions 31 and 91, flow channel connection portions 73 and 74 having the same configuration as the flow channel connection portions 30, 40, and 90. Is provided. The flow path connection portion 73 and the flow path connection portion 74 are provided on the same plane of the outer surface of the casing 72 of the bypass cartridge 70, and the distance between the flow path connection portion 73 and the flow path connection portion 74 is determined by the device connection. By making it the same as the distance between the part 31 and the device connection part 91, the connection between the flow paths 3 and 9 and the bypass cartridge 70 and the connection from the bypass cartridge 70 to the cell trapping device 1 can be smoothly switched. it can.

  Similar to the cell trapping device 1, the bypass cartridge 70 is fixed on the mount 19 of the moving device 17A. In addition to the drive motor 20A for moving in the Y-axis direction, the moving device 17A in the cell trapping apparatus 100A shown in FIG. 11 includes a drive motor 20B so as to be movable in the X-axis direction. By moving the bypass cartridge 70 and the cell capturing device 1 by such a moving device 17A, the flow path 3, the bypass cartridge 70, and the flow path 9 are connected before the cell capturing process in the cell capturing apparatus 100A is started. Then, the bubbles in the flow path 3 can be mainly discharged by circulating the treatment liquid. Thereafter, the bypass cartridge 70 is removed from the flow path 3 and the flow path 9 by the moving device 17A, and the cell capture device 1 is connected to the flow paths 3, 4, and 9 to start processing related to cell capture. It is possible to prevent outside air from being mixed into the inside of the cell trapping device, and to further suppress the generation of bubbles inside the processing solution or the test solution in the cell trapping device. it can.

In the cell capture device 100A, the bypass cartridge 70 and the cell capture device 1 are fixed on the same mounting base 19 and moved together. However, the bypass cartridge 70 is connected to the cell capture device 1 and the cell capture device 1A. May be configured to be moved by different moving means (second moving means).
(Modification-5)

  Next, a second modification of the configuration of the cell trapping device 100 will be described. The cell capture device 100B shown in FIG. 12 is obtained by changing the following points compared to the cell capture device 100A shown in FIG. That is, the object to be moved in the moving device 17B is changed.

  Specifically, the moving device 17B in the cell trapping device 100B includes a mounting base 19 that can move in the X-axis direction by driving of the drive motor 20B, and mounting bases 191 to 193 that can move in the Y-axis direction. Among these, the cell trapping device 1 and the bypass cartridge 70 are fixed on the mount 19. The device connection part 31 of the flow path 3 is fixed to the mounting base 191, the device connection part 41 of the flow path 4 is fixed to the mounting base 192, and the device connection part 91 of the flow path 9 is fixed to the mounting base 193. Yes. When the device connection portions 31, 41, 91 are respectively fixed to the mounting bases 191 to 193, the mounting bases 191 to 193 can be moved independently of each other. Therefore, the device to the cell trapping device 1 or the bypass cartridge 70 is used. The connection parts 31, 41, 91 can be connected or disconnected individually.

  Thus, the mechanism of the moving device and the object to be moved by the moving device can be changed as appropriate.

  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, the configuration of each part of the cell trapping apparatus 100 is not limited to the above, and for example, the shape of the cell trapping device 1 can be changed as appropriate.

  Moreover, you may add apparatuses, such as a deaeration apparatus, in addition to each part of the cell capture apparatus 100 demonstrated in the said embodiment as needed.

  In addition, the cell capturing device and the operation method of the device of the present invention are useful for capturing cells in blood (red blood cells, white blood cells, platelets, circulating blood cancer cells, etc.). Since (CTC) is more difficult to deform than leukocytes in blood, it is useful when separating and capturing circulating cancer cells in blood from blood containing leukocytes.

DESCRIPTION OF SYMBOLS 1 ... Cell capture device, 3, 4, 9 ... Flow path, 5 ... Treatment liquid storage container, 10 ... Test liquid storage container, 17 ... Moving apparatus, 30, 30A, 30B, 30C, 40, 90 ... Flow path connection Part, 31, 41, 91 ... device connection part, 70 ... bypass cartridge, 100, 100A, 100B ... cell trapping device.

Claims (7)

An introduction flow path for introducing a test liquid or a processing liquid for processing cells in the test liquid into the inside, and a discharge flow for discharging the introduced test liquid or the processing liquid to the outside A passage and a plurality of through holes are formed in the thickness direction, and the test liquid or the treatment liquid is disposed on the flow path between the introduction flow path and the discharge flow path so as to pass through the through hole. A cell capture device comprising: a filter; and a housing in which the introduction channel, the discharge channel, and the filter are provided,
The introduction flow path and the discharge flow path each have a flow path connection portion that can connect an external flow path for allowing the test liquid or the processing liquid to flow between the outside and the outside on the surface of the housing. Have
The flow path connecting portion includes an on-off valve that opens and closes the flow of the test liquid or the processing liquid,
When the external flow path is connected to the flow path connection portion, the on-off valve opens the flow of the test liquid or the processing liquid, and the external flow path is connected to the flow path connection portion. A cell-capturing device that closes the flow of the test liquid or the treatment liquid when a flow path is not connected.   The cell trapping device according to claim 1, wherein the on-off valve is configured by an elastic member provided so as to seal the introduction flow path or the discharge flow path at the flow path connection portion.   The on-off valve includes a sealing member on a flat plate provided so as to seal the introduction flow path or the discharge flow path in the flow path connection portion, and an elastic member that supports the sealing member so as to be movable. The cell capturing device according to claim 1, comprising:   The cell capture device according to any one of claims 1 to 3, wherein a flow path connection portion of the introduction flow path and the discharge flow path is provided on the same plane of the housing. The cell capture device according to any one of claims 1 to 4,
A test solution storage container for storing the test solution;
An external flow path that connects the test liquid storage container and the introduction flow path of the cell capture device, and has a device connection portion that can be connected to the flow path connection section of the introduction flow path A liquid flow path;
A processing liquid storage container for storing the processing liquid for processing the cells captured by the filter by passing through the filter;
A treatment liquid flow having an external flow path for connecting the treatment liquid storage container and the introduction flow path of the cell capture device, the device connection section being connectable to the flow path connection section of the introduction flow path Road,
An external flow path connected to the discharge flow path of the cell trapping device, and a system discharge flow path having a device connection portion connectable to the flow path connection portion of the discharge flow path;
A selection means for selecting a liquid to be supplied to the cell capture device from the test liquid and the treatment liquid;
Based on the selection result of the selection means, a liquid feeding means for supplying a test liquid from the test liquid storage container or a treatment liquid from the treatment liquid storage container to the cell capturing device;
A cell capture device comprising:   The cell capturing apparatus according to claim 5, further comprising first moving means for movably supporting the cell capturing device.   A bypass flow supported by the second moving means and having one end side connectable to the device connection part of the processing liquid flow path and the other end side connectable to the device connection part of the extra-system discharge flow path The cell capturing device according to claim 6, further comprising a path.

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