JP6124285B2 - Circulating cancer cell trap - Google Patents

Description

  The present invention relates to a circulating cancer cell capturing device for capturing circulating cancer cells that have entered into blood for use in testing for metastatic cancer or evaluating anti-cancer using a filter.

  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, riding on blood, spreading throughout the body, and cancer cells 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.

  As a conventional technique for capturing circulating cancer cells that are cancer cells in blood that cause metastasis, for example, as shown in Patent Document 1, a method of capturing cancer cells in blood with a filter is known. Patent Document 1 discloses a manufacturing method using a semiconductor technology of a filter, the shape of a cell unit containing a filter, and the structure of a flow channel for flowing blood and processing liquid. It is intended to capture cancer cells by flowing blood containing cells. Cells existing on the filter are identified as cancer cells by staining. In Patent Documents 2 and 3, a cell dispersion liquid 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.

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

  However, when the devices described in Patent Documents 1 to 3 are used, it is very troublesome to capture cancer cells in blood and start measurement, and it is difficult to process a large number of specimens. In addition, when air is mixed into the flow path of the apparatus, there is a concern that the efficiency of sample processing is significantly reduced and the measurement accuracy is further reduced.

  The present invention has been made in view of the above, and provides a circulating cancer cell capturing device capable of suppressing the mixing of air into the flow path and performing the capturing operation of circulating cancer cells in blood more efficiently. The issue is to provide.

  In order to achieve the above object, a circulating cancer cell capturing device according to the present invention includes a blood storage container for storing blood; and a treatment liquid for processing the blood or circulating cancer cells in the blood. An introduction channel, a discharge channel for discharging the blood or the treatment liquid to the outside, and a plurality of through holes formed in the thickness direction between the introduction channel and the discharge channel. A filter unit comprising a filter disposed on the flow path of the blood or the treatment liquid so as to pass through the through-hole; a blood flow connecting the blood storage container and the introduction flow path of the filter unit A processing liquid storage container for storing a processing liquid for processing circulating cancer cells captured by the filter by passing through the filter; and the introduction flow path of the processing liquid storage container and the filter unit And connect A treatment liquid flow path; selection means for selecting a liquid to be supplied to the filter unit from the blood and the treatment liquid; blood from the blood storage container or the treatment based on a selection result of the selection means; Liquid feeding means for supplying the processing liquid from the liquid storage container to the filter unit; degassing means for capturing air in the blood or in the processing liquid and discharging it to the outside upstream of the filter; It is characterized by providing;

  According to the above circulating cancer cell capturing device, the liquid to be supplied to the filter unit is selected by the selecting means, and the blood or the processing liquid is supplied to the filter unit by the liquid feeding means based on the result. It becomes possible to advance the capture | acquisition work of the circulating cancer cell in the blood efficiently compared with the past. Furthermore, by providing a deaeration means for capturing the air in the blood and discharging it to the outside upstream from the filter, it is possible to avoid the blood in a state where the air is mixed from passing through the filter. Therefore, it is possible to suppress a reduction in processing efficiency due to air mixing and maintain high measurement accuracy.

  Here, as a configuration that effectively exhibits the above-described operation, specifically, the degassing unit includes an air reservoir provided on the blood channel or the treatment liquid channel, and the blood channel Alternatively, the upstream side of the air reservoir in the processing liquid channel supplies the blood or the processing liquid into the air reservoir from the upper part of the air reservoir, and the air in the blood channel or the processing liquid channel A mode in which the blood or the treatment liquid is discharged from the lower part of the air pool is provided on the downstream side of the pool.

  Here, as another configuration that effectively exhibits the above action, specifically, the deaeration means is connected so that one end thereof communicates with the blood flow path or the treatment liquid flow path. The first and second deaeration channels are attached so as to be inclined so that the position gradually becomes higher toward the other end, and the other end is an open end. Can be mentioned.

  The deaeration means is connected to the other end of the first deaeration channel and stores a blood from the first deaeration channel, and on the blood channel or the processing liquid channel A first flow path valve provided on the upstream side of the connection position with the first degassing flow path to open and close the blood flow path or the treatment liquid flow path. It is also possible to adopt a mode in which the liquid feeding means is driven with the passage valve closed, and the blood stored in the stored solution is fed to the filter unit.

  Moreover, the said deaeration means can also be set as the aspect comprised including the 2nd deaeration flow path branched and extended upwards from the said introduction flow path of the said filter unit.

  The deaeration unit may further include a vibrator using a piezoelectric element installed so as to be in contact with the filter unit.

  In addition, a plurality of the processing liquid storage containers are provided, and the processing liquid flow paths are connected to a plurality of upstream flow streams connected to a first end which is one end with respect to each of the plurality of processing liquid storage containers. A downstream channel that connects the channel, any one of the second ends that are different from the first ends of the plurality of upstream channels, and the introduction channel of the filter unit; And a selection valve that can be connected to the downstream flow path by selecting any one of the second end portions of the plurality of upstream flow paths, The selection valve can be controlled based on the selection result of the selection means.

  In addition, a plurality of the processing liquid storage containers are provided, and the processing liquid flow path includes a suction nozzle that sucks the processing liquid in the processing liquid storage container, and a processing liquid sucked by the suction nozzle. A nozzle pipe for supplying the introduction flow path; and a position changing means for changing the position of the suction nozzle with respect to the processing liquid storage container. The liquid feeding means is based on a selection result of the selection means. Thus, it is possible to move the suction nozzle by the position changing means and suck the processing liquid in the processing liquid storage container to perform liquid feeding.

  Here, the position changing unit may change the position of the suction nozzle with respect to the processing liquid storage container by moving the processing liquid storage container. Further, the position changing means may change the position of the suction nozzle with respect to the processing liquid storage container by moving the suction nozzle. Further, the position of the suction nozzle with respect to the processing liquid storage container may be changed using a linearly moving table that moves only along a predetermined direction.

  The suction nozzle may further include at least one of a cleaning mechanism that cleans an area in contact with the processing liquid stored in the processing liquid storage container and an exchange mechanism that replaces the area.

  In addition, a plurality of the filter units are provided, and the blood flow path and the processing liquid flow path are provided corresponding to the plurality of filter units, respectively, and the selection unit supplies the plurality of filter units. Each of the plurality of filter units is configured to supply the blood from the blood storage container or the processing liquid from the processing liquid storage container to each of the plurality of filter units based on the selection result of the selection means. It is good also as an aspect supplied with respect.

  In addition, a plurality of liquid feeding units may be provided so as to correspond to the plurality of filter units.

  Furthermore, it is good also as an aspect further equipped with the circulating cancer cell specific | specification means which performs identification of the circulating cancer cell captured on the said filter, and count of quantity.

  Further, as the circulating cancer cell specifying means, an image acquiring means for acquiring a circulating cancer cell image on the filter is provided, and the identification and quantity of the circulating cancer cells based on the image acquired by the image acquiring means It is also possible to adopt a mode in which counting is performed.

  Moreover, it is good also as an aspect further equipped with the output means which outputs the image acquired by the said image acquisition means as said circulating cancer cell identification means.

  Furthermore, it is also possible to adopt a mode further comprising circulating cancer cell specifying means for identifying and counting the number of circulating cancer cells captured on the filter corresponding to each of the plurality of filter units.

  The plurality of filter units may be integrated.

  Furthermore, the liquid feeding means may be configured to include a liquid feeding pump. In that case, the liquid feeding pump may be a peristaltic pump.

  Further, the liquid feeding means is configured to include a pressure-variable decompression tank connected to the flow path and a flow meter for measuring the pressure in the flow path, and according to the measured value of the flow meter, It is good also as an aspect which changes the pressure of the said pressure reduction tank.

  ADVANTAGE OF THE INVENTION According to this invention, the circulating cancer cell capture device which suppresses mixing of the air in a flow path and can perform the capture | acquisition operation | work of the circulating cancer cell in blood more efficiently is provided.

It is a figure explaining the structure of the circulating cancer cell capture device which concerns on this embodiment. It is a figure explaining the filter unit in the circulating cancer cell capture device of FIG. It is a figure explaining the modification of a circulating cancer cell capture device. It is a figure explaining the modification of a circulating cancer cell capture device. It is a figure explaining the modification of a circulating cancer cell capture device. It is a figure explaining the modification of a circulating cancer cell capture device. It is a figure explaining the modification of a circulating cancer cell capture device. It is a figure explaining the modification of a circulating cancer cell capture device. It is a figure explaining the modification of a circulating cancer cell capture device. It is a figure explaining the modification of a circulating cancer cell capture device. It is a figure explaining the nozzle used for the circulating cancer cell capture device of FIG. It is a figure explaining the circulating cancer cell capture | acquisition system containing a circulating cancer cell specific means. It is a figure explaining the modification of a circulating cancer cell capture system. It is a figure explaining the deaeration means of the circulating cancer cell capture device. It is a figure explaining the modification of the deaeration means of the circulating cancer cell capture device. It is a figure explaining the modification of the deaeration means of the circulating cancer cell capture device. It is a figure explaining the modification of the deaeration means of the circulating cancer cell capture device. It is a figure explaining the modification of the suction means of a circulating cancer cell capture device. It is a figure explaining the modification of the suction means of a circulating cancer cell capture 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.

(Circulating cancer cell capture device)
FIG. 1 is a diagram illustrating the configuration of a circulating cancer cell capturing device according to this embodiment. A circulating cancer cell capturing device is a device that captures circulating cancer cells contained in blood by filtering blood as a test solution using a filter. In addition, the circulating cancer cells captured by the filter are stained using a treatment solution such as a staining solution, whereby the circulating cancer cells are identified and the number of cancer cells is counted.

  As shown in FIG. 1, a circulating cancer cell capturing device 100 is provided with a filter unit 1 in which a filter for capturing circulating cancer cells is provided, and for supplying a treatment liquid (reagent) to the filter unit 1. A flow path 3 (treatment liquid flow path / downstream flow path) made of a soft tube and a flow path 4 (blood flow path) made of a soft tube for supplying blood to the filter unit 1 are provided. Different processing liquids (reagents) are placed in the plurality of processing liquid storage containers 5 on the upstream side of the flow path 3. Examples of the treatment liquid charged into the treatment liquid storage container 5 include a staining liquid for staining circulating cancer cells, a washing liquid for washing circulating cancer cells captured by a filter, a rinsing liquid, and the like. 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 path / upstream flow path). These flow paths are connected to the selection valve 8, and the processing liquid (processing liquid storage container 5) to be connected to the flow path 3 is selected by rotating the selection valve 8 (selection valve). The

  Connected to the flow path 4 connected to the filter unit 1 is a blood storage container 10 in which blood containing circulating cancer cells is stored as a test solution. The filter unit 1 is configured to supply either one of the processing liquid and blood instead of supplying the processing liquid and blood at the same time. Switching is performed by valves 12 and 13 attached to 3 and 4 respectively. For example, when supplying blood to the filter unit 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.

  Further, when any one of the processing liquid and the blood is supplied to the filter unit 1, the liquid is supplied by being sucked by a pump 14 (supply means) provided downstream of the filter unit 1. . 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, a liquid such as a reagent or blood flows through the flow path 3 or the flow path 4 in the direction toward the filter unit 1 and is supplied to the filter unit 1. The liquid that has passed through the filter unit 1 flows into the waste liquid tank 16. With these structures, circulating cancer cells in the blood are captured by a filter provided on the flow path in the filter unit 1, and the circulating cancer cells are stained with a staining solution.

  The above-described control of each part is performed by control by the control part 48 (selection means, liquid feeding means). Specifically, the selection valve 8 is controlled by the selection valve driver 49 based on an instruction from the control unit 48. Further, the valves 12 and 13 are controlled by two valve drivers 50 connected thereto. Also, the drive of the pump 14 is controlled by the pump driver 51. For example, by changing the number of rotations of the pump, the flow rate is variable, and the determined optimal flow rate is supplied to the filter unit 1, Capturing circulating cancer cells in the blood and staining the captured circulating cancer cells can be performed under an optimal flow rate condition. The control unit 48 is provided with a program input function for inputting a program that enables control such as driving / stopping for each of the selection valve 8, the valves 12, 13, and the pump 14. As described above, a drive mechanism for operating each device in sequence is added according to the program.

  Next, the filter unit 1 will be described with reference to FIG. 2A is a top view of the filter unit, and FIG. 2B is an additional view of deaeration means attached to the flow path 4 with respect to the IIB-IIB arrow view of FIG. 2A. Is.

  The filter unit 1 is configured such that a filter 57 having a plurality of through holes 61 is sandwiched between an upper container 58 and a lower container 59 in which a space is formed. The filter 57 is made of, for example, metal and has a plurality of through holes 61 formed in the thickness direction. The upper container 58 of the filter unit 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. An upper space 62 that is formed and communicates with the flow paths 3A and 4A and is formed above the filter 57 is provided. That is, in the present embodiment, the “introduction channel” refers to the channels 3A and 4A provided in the filter unit 1 among the channels. “Blood flow path” and “treatment liquid flow path” refer to the flow paths 3 and 4 connected to the outside of the filter unit 1.

  The lower container 59 communicates with the lower space 63 and the lower space 63 formed below the filter 57 and having a central portion deeper than the peripheral edge portion. A flow path 59A (discharge flow path) for discharging to the outside is provided. The through-hole 61 provided in the filter 57 sandwiched between the upper container 58 and the lower container 59 is set to a size that prevents the circulating cancer cells 65 serving as a capture target from passing therethrough. Thereby, when blood passes through the through hole 61, the circulating cancer cells 65 are captured on the filter 57. Here, the washing and staining for detecting the captured circulating cancer cells are performed by supplying the washing solution and the staining solution from the flow path 3 of the tube connected to the flow path 3A after the blood filtering is completed. . By this operation, the circulating cancer cells are captured using the filter 57 and stained with the treatment liquid. The filter unit 1 is removed from the flow paths 3 and 4 for identification of circulating cancer cells and measurement of the number of circulating cancer cells as required.

  A deaeration means 150 for removing air mixed in the flow path 4 is attached on the flow path 4 for supplying blood to the filter unit 1. This deaeration means is a characteristic part of the circulating cancer cell capture device of the present invention, and details thereof will be described later.

  In the circulating cancer cell capturing apparatus 100 described above, the control of the control unit 48 allows a series of processes of capturing circulating cancer cells in the blood by the filter 57 of the filter unit 1 and washing and staining the captured circulating cancer cells. It can be done as an action. Therefore, the operation of capturing circulating cancer cells in blood can be performed more efficiently than in the past.

(Modification of circulating cancer cell capture device)
Next, a modified example of the circulating cancer cell capturing device will be described with reference to FIGS.

(Modification 1)
In the circulating cancer cell capturing device 100 </ b> A shown in FIG. 3, the filter unit 1 that houses the filter 57 is connected to the flow paths 3 and 4. A plurality of processing liquid storage containers 5 in which reagents such as a staining liquid, a cleaning liquid, and a rinsing liquid are stored are provided upstream of the flow path 3. A flow path 6 is attached to each of the plurality of processing liquid storage containers 5. Each flow path 6 is connected to the flow path 3 connected to the filter unit 1, but a selection valve 18 is provided on the upstream side of the position where each of the plurality of flow paths 6 joins the flow path 3. It has been. That is, the selection valve 18 is disposed so as to correspond to the plurality of processing liquid storage containers 5 and the flow paths 6 attached thereto. The liquid to be supplied to the flow path is selected by opening and closing the selection valve 18. Further, similarly to the circulating cancer cell capturing device 100 of FIG. 1, a valve 13 is also attached to the flow path 4 on the blood supply side, and the liquid supplied to the filter unit 1 is opened and closed by these valves. Selected. The liquid selected by opening and closing the valve is supplied by driving the pump 14.

  A plurality of valves 18 attached to each of the plurality of processing liquid storage containers 5 and a valve 13 attached to the flow path 4 for supplying blood are respectively connected by a valve driver 50 connected to the control unit 52. Be controlled. The driving of the pump 14 is controlled by a pump driver 51. Accordingly, the control unit 52 opens and closes the valves 13 and 18 and drives the pump 14, and a desired liquid is supplied to the filter unit 1. Therefore, similarly to the circulating cancer cell capturing device 100 shown in FIG. 1, the circulating cancer cell capturing device 100A shown in FIG.

(Modification 2)
The circulating cancer cell capturing device 100B shown in FIG. 4 is different from the circulating cancer cell capturing device 100 of FIG. 1 in the following points. That is, two selection valves that are connected via the flow path 6 to each of the processing liquid storage containers 50 in which various reagents are stored are provided. The flow path 6 from the processing liquid storage container 5 to the selection valves 24 and 25 has a structure that branches in the middle. For this reason, it has a structure in which one set of processing liquid storage container 5 can be shared and used by two filter cell units. The selection valves 24 and 25 are driven independently. Further, the flow paths 3 and 3 ′, the filter units 1 and 1 ′, the blood storage containers 10 and 10 ′, and the pumps 14 and 14 ′ in the subsequent stage of the selection valves 24 and 25 are provided separately, and can be operated independently. It has a possible structure. In addition, although the structure which each arrange | positioned the waste liquid tanks 16 and 16 'corresponding to the filter units 1 and 1' is shown in FIG. 4, it is also possible to share and arrange one. Similarly to the circulating cancer cell capturing devices 100 and 100B, the selection valves 24 and 25, the blood storage containers 10 and 10 ′, and the pumps 14 and 14 ′ are controlled by the control unit. As described above, the circulating cancer cell capturing apparatus 100B shown in FIG. 4 can capture the circulating cancer cells in the blood more efficiently as in the circulating cancer cell capturing apparatus 100 shown in FIG. Furthermore, in the circulating cancer cell capturing device 100B of FIG. 4, two specimens can be processed simultaneously.

(Modification 3)
In the circulating cancer cell capturing device 100C shown in FIG. 5, unlike the circulating cancer cell capturing devices 100, 100A, 100B described above, there is one flow path for supplying a liquid to the filter unit. A reagent for discriminating blood or circulating cancer cells as a test solution is supplied into the flow path. In the circulating cancer cell capturing device 100C, the flow path 3 is connected to the filter unit 1 that houses the filter 57 therein. A supply pipe 32 (nozzle pipe) for supplying the reagent to the flow path 3 is attached to the upstream side of the flow path 3. A pump 14 is disposed on the downstream side of the filter unit 1, and has a structure capable of flowing the solution supplied from the supply pipe 32 and passing through the flow path 3 and the filter unit 1 toward the waste liquid tank 14. Yes.

  The blood storage container 11 in which blood is stored and the treatment liquid storage container 5 in which a reagent is stored are arranged in an annular shape centering on a rotary table 43 (position changing means) of the movable pipette 40, and the movable pipette 40 is arranged on the rotary table 43. , The liquid in the movable pipette 40 is discharged by sucking each liquid from the blood storage container 11 and the processing liquid storage container 5 and discharging the sucked liquid on the supply pipe 32. It is supplied to the supply pipe 32 and sent to the filter unit 1 through the flow path 3. Further, a nozzle replacement mechanism 45 (exchange mechanism) for replacing the nozzle 44 (suction nozzle) of the movable pipette 40 is provided on the ring on which the blood storage container 11 and the treatment liquid storage container 5 are provided. The nozzle replacement mechanism 45 has a function of attaching to the movable pipette 40 and a nozzle corresponding to the liquid that the movable pipette 40 supplies to the supply pipe 32 while rotating around the axis. Instead of the replacement mechanism, a cleaning mechanism for cleaning the nozzle to which the processing liquid or the like adheres may be provided.

  In the circulating cancer cell capturing device 100 </ b> C described above, the above-described operation is performed according to an instruction from the control unit 53. That is, the pump 14 is controlled by the pump driver 51 in accordance with an instruction from the control unit 53, and the rotation speed is changed to control the start / stop of the pump 14 and to change the flow rate. Further, the rotational position control and the vertical position control of the movable pipette 40 are controlled based on an instruction from the control unit 53 via the rotational drive driver 54 and the vertical drive driver 55. Further, the nozzle replacement mechanism 45 is controlled by the control unit 53 via the nozzle replacement mechanism driver 56. That is, in the circulating cancer cell capturing device 100C, the control of the pump 14, the movable pipette 40, and the nozzle replacement mechanism 45 by the control unit 53 enables the control of automatically capturing circulating cancer cells in the blood. As described above, the circulating cancer cell capturing device 100C shown in FIG. 5 can perform the capturing operation of circulating cancer cells in blood more efficiently.

(Modification 4)
In the circulating cancer cell capturing device 100D shown in FIG. 6, a movable pipette 73 is used in the same manner as the movable pipette 40 in which the circulating cancer cell capturing device 100C shown in FIG. Other configurations are the same as those of the circulating cancer cell capturing apparatus 100. That is, the flow path 4 from the blood storage container 10 and the flow path 3 from the reagent supply pipe 67 are connected to the filter unit 1. The reagent supply pipe 67 is provided separately from the processing liquid storage container 5 that stores a reagent such as a staining liquid. A pump 14 for flowing liquid from the flow paths 3 and 4 to the filter unit 1 and a drain tank 16 are disposed downstream of the filter unit 1. Valves 70 and 71 are attached to the flow paths 3 and 4, respectively. Therefore, when blood is supplied to the filter unit 1, the valve 70 is closed and the valve 71 is opened. Supply to the reagent supply pipe 67 of the staining solution or the like placed in the processing solution storage container 5 is performed using a pipette 73 (a suction nozzle) that is located above these and is movable in the horizontal direction. The pipette 73 has a structure that can be moved in the vertical direction as well. The reagent is selected by driving the pipette moving mechanism 75, and the circulating cancer cells captured on the metal filter in the filter unit 1 are washed and stained. Perform the work. These pipette moving mechanism 75 (position changing means / straight moving table) and pump 14 are controlled by a control unit (not shown) so that the circulating cancer cells can be captured and stained efficiently in a predetermined process. Can do.

(Modification 5)
In the circulating cancer cell capturing device 100E shown in FIG. 7, compared with the circulating cancer cell capturing device 100D shown in FIG. 1 is different in that a flow path 77 connected to 1 is provided. That is, the suction port 78 for supplying the reagent is movable in the horizontal direction and the vertical direction by the suction port moving mechanism 81, and the cartridge 82 provided at the tip of the suction port 78 is used as the reagent in the processing liquid storage container 5. Contact and aspirate reagent from here. The reagent is supplied to the filter unit 1 via the flow path 77 as it is. The supply of blood from the blood container 10, the driving of the pump 14, the opening and closing of the valves 70 and 71 on the flow path, and the like are the same as those of the circulating cancer cell capturing device 100D. In the case where contamination is a problem because the same suction port 78 is used for a plurality of processing liquid storage containers 5 in which different reagents are stored, a cartridge that replaces the cartridge 82 for each suction step An exchange mechanism can be attached. Further, the driving of the mouthpiece moving mechanism 81 and the pump 14 and the opening and closing of the valves 70 and 71 are controlled by the control unit, so that the circulating cancer cells can be captured and stained efficiently in a predetermined process. it can.

(Modification 6)
In the circulating cancer cell trapping apparatus 100F shown in FIG. 8, the circulating cancer cell trapping apparatus 100E shown in FIG. 7 accommodates the reagent as compared with the configuration in which the mouthpiece 78 is movable by the mouthpiece moving mechanism. The difference is that the reagent supply base 85 in which a plurality of processing liquid storage containers 5 are arranged is movable. That is, in the circulating cancer cell capturing apparatus 100F shown in FIG. 8, the reagent supply base 85 is attached on the reagent moving stage 86, and the reagent moving stage 86 enables horizontal and vertical movement. On the other hand, the suction port 84 with the cartridge 89 attached to the tip is attached to a fixed base (not shown). The reagent supplied to the filter unit 1 is introduced into the flow path 83 from the suction port 84 by contacting the cartridge 89 of the suction port 84 by moving the reagent moving stage 86 under the control of a control unit (not shown). Is done. When contamination of the suction port 84 becomes a problem, a cartridge replacement mechanism that replaces the cartridge 89 of the suction port for each suction process can be attached. The driving of the reagent moving stage 86 and the pump 14 and the opening and closing of the valves 70 and 71 are controlled by the control unit, so that the circulating cancer cells can be captured and stained efficiently in a predetermined process. .

(Modification 7)
In the circulating cancer cell capturing device 100G shown in FIG. 9, a plurality of sets of the circulating cancer cell capturing device 100F shown in FIG. 8 are arranged in parallel to improve the processing capability. Specifically, except that the drain tank 16 is combined into one, the configuration of the circulating cancer cell capturing device 100F shown in FIG. 8, that is, the reagent supply base 85 attached to the movable reagent moving stage 86 is shown. The processing liquid storage container 5 is attached on the top. The supply of blood and reagents to the four filter units 1 arranged in parallel can be performed independently. As described above, in the circulating cancer cell capturing apparatus 100G, it is possible to simultaneously handle a plurality of specimens, up to four specimens, and to increase the number of processes. In addition, the number arrange | positioned in parallel can be changed suitably.

(Modification 8)
In the circulating cancer cell capturing apparatus 100H shown in FIG. 10, a multi-filter unit 1B in which a plurality of metal filters 1A are arranged in a flat plate and a suction port 107 is attached to each of the plurality of metal filters 1A is used. Yes. Independent flow paths 108 are connected to each of the plurality of metal filters 1A so that they do not mix with each other. Further, the plurality of flow paths can all be sucked by driving the pump 109.

  The multi-filter unit 1B is attached to the vertical moving table 110 and has a structure that can move in the vertical direction. A rotary moving table 112 is provided below the multi-filter unit 1B. In addition to the blood supply unit 11 in which a plurality of blood storage containers 10 are arranged corresponding to the arrangement of the plurality of metal filters 1A on the rotary moving table 112, a plurality of treatment liquid storage containers 5 are formed of the plurality of metal filters 1A. Reagent units 7 arranged corresponding to the arrangement are arranged. A plurality of types of reagent units 7 are arranged according to the type of reagent.

  A multi-filter unit 1B to which a plurality of metal filters 1A are attached will be described with reference to FIG. FIG. 11 is a schematic cross-sectional view of the multi-filter unit 1B and shows an example in which four filters 1A are arranged. An upper space 114 and a lower space 115 for flowing a fluid are arranged above and below the filter 1A, and the processing liquid or the like flows from the lower side to the upper side after passing through the metal 1 filter 1A after being sucked by the suction port 107. . Here, when the blood passes through the metal filter 1A, circulating cancer cells in the blood are captured. In addition, in FIG. 11, the structure which the suction port 107 was attached to the multi filter unit 1B via the tube 117 is shown. When this structure is adopted, it is not necessary to fix the filter unit 1B to the vertical movement table 110 shown in FIG. By adopting this structure, the moving parts can be miniaturized, and the circulating cancer cell capture device can be miniaturized.

  By having the above configuration, it is possible to simultaneously process samples as many as the number of metal filters 1A. Therefore, it is possible to efficiently capture and stain circulating cancer cells and increase the number of treatments.

(About observation of circulating cancer cells after capture)
Circulating cancer cells captured by the above-described circulating cancer cell capturing device are advanced to an observation process for quantity measurement and the like. FIG. 12 shows an example of an observation apparatus 200 for observing captured circulating cancer cells. The observation apparatus 200 includes a microscope 96, a stage 97, a monitor 101 (output means), a computer 102, and a stage driver 103.

  The stained circulating cancer cells are visually recognized by mounting the filter unit 1 that has been stained after the circulating cancer cells in the blood are captured by the circulating cancer cell capturing device described above on the stage 97 of the microscope 96. be able to. A motor 98 is attached to the stage 97 on which the filter unit 1 is mounted and is movable. Thereby, the entire surface of the filter in the filter unit 1 can be scanned in the field of view of the microscope 96. In addition to viewing with the eyepiece lens 99, the microscope 96 can be projected on the monitor 101 by attaching a camera. In addition, by transmitting image information captured by a camera (image acquisition means) to the computer 102, it is possible to determine and evaluate the characteristics of the circulating cancer cells using the computer 102. In addition, since the computer 102 and the stage 97 are linked via the stage driver 103, the position of the cancer cell can be specified on the filter. With the above configuration, the observation apparatus 200 according to the present embodiment can automatically count the number of captured circulating cancer cells.

  Furthermore, the example which combined the observation apparatus 200 and the circulating cancer cell capture apparatus in FIG. 13 is shown. In the circulating cancer cell observation system 200A, five circulating cancer cell capturing devices 100 (which may be modified circulating cancer cell capturing devices 100A to 100H) are arranged, and the circulation shown in FIG. One cancer cell observation device 200 is connected. Here, the observation apparatus 200 takes in the filter unit that has captured the circulating cancer cells from each circulating cancer cell capturing apparatus 100, and identifies and counts the circulating cancer cells captured by the metal filter in the filter unit. Has been. Here, in consideration of the time required for capturing the circulating cancer cells in the circulating cancer cell capturing device 100 and the processing time for counting the circulating cancer cells in the observation device 200, the connection is made to the observation device 200. The number of circulating cancer cell capturing devices 100 is determined. In the circulating cancer cell observation system 200A, since the capture time of the circulating cancer cells is about five times longer than the time for identifying and counting the captured cells, the capture device is compared with one observation device. It is structured to connect 5 units. As a result, a circulating cancer cell capture and observation system can be configured that has no waiting time for the apparatus and is lean.

(Deaeration means)
Next, the deaeration means attached to the filter unit, which is one of the features forming the present invention, will be described with reference to FIG. FIG. 14 is an enlarged view of the vicinity of the filter unit 1 in the circulating cancer cell capturing apparatus 100 according to the present embodiment.

  In FIG. 14, the deaeration means provided in the flow path 4 connected to the filter unit 1 will be described. One end portion is attached to the deaeration means so as to branch from the flow path 4, and the branch flow path (the first flow path is attached so as to be inclined upward toward the other end side). A degassing flow path) 131, a container 132 (storage container) attached to the other end of the flow path 131, and a valve 133 provided above the container. By attaching the branch flow path 131 and the container 132 to the upper side with respect to the flow path 4, the bubbles 67 flowing in the flow path 4 move to the container 132 side branched from the flow path 4. Then, by opening and closing the valve 133 attached to the container 132, the air flowing in the flow path 4 as bubbles is discharged out of the container 132. That is, when the valve 133 is opened, the end of the separation channel 131 becomes an open end, and the bubbles 67 are discharged from the end.

  Thus, by providing the deaeration means, the bubbles 67 in the flow path 4 can be discharged to the outside, the state in which air remains in the flow path can be reduced, and the measurement accuracy can be improved. In addition, the deaeration means in the circulating cancer cell capturing apparatus 100 in the present embodiment aims to remove air or the like mixed in the flow path and flowing together with blood or treatment liquid. When air or the like enters the flow path, the volume of liquid in the flow path decreases (the volume of liquid that can be transported with respect to the flow rate decreases), or when air enters even at the same pump speed Is affected by the fact that it is difficult to move the liquid at the assumed flow rate, resulting in the problem that the liquid (for example, blood) in the flow path cannot be moved as expected, resulting in circulation. Even if the number of cells is counted after capturing the cancer cells, it is difficult to obtain an accurate value. Therefore, by providing the deaeration means in this way, air mixed in the flow path 4 can be removed, and the circulating cancer cells can be captured and measured with higher accuracy. Although FIG. 14 shows a configuration in which a container 132 serving as a degassing unit is connected to the flow channel 4, a degassing unit may be provided in the flow channel 3 for supplying the reagent. It is also possible to provide deaeration means in both the flow path 3 and the flow path 4.

(Modification of deaeration means)
Next, several examples of modified examples of the deaeration means will be described. First, the 1st modification of a deaeration means is demonstrated using FIG. In FIG. 15, the deaeration means provided on the flow path 4 connected to the filter unit 1 will be described. The deaeration means 150A includes a container 134 that functions as an air reservoir. The flow path 4 supplies blood into the container from the upper part of the container 134 on the upstream side of the container 134. On the downstream side, blood is discharged from the lower side of the container 134 with respect to the flow path 4C. At this time, the bubbles 67 flowing in the flow path 4 are mixed with the gas in the vicinity of the water surface of the container or in the container 134 when discharged from the flow path 4 to the container 134. When the valve 133 is opened, the upper part of the container 134 is opened and air bubbles are discharged therefrom. Then, the blood from which the bubbles 67 have been removed is sent to the downstream channel 4C. By having such a configuration, the bubbles in the flow path 4 can be discharged to the outside, the state in which air remains in the flow path can be reduced, and the measurement accuracy can be improved.

  Next, a second modification of the deaeration means will be described with reference to FIG. In FIG. 16, the filter unit 1 itself is provided with a pipe and a valve that serve as a second degassing flow path, and functions as a degassing means. Specifically, as shown in FIG. 16, the filter unit 1 is arranged so that the filter 57 is in the vertical direction, and the end on the downstream side of the introduction flow path 4A is the upper side when the filter unit 1 is arranged. A pipe 141 (second deaeration channel) extending upward and a valve 142 are attached. As a result, the bubbles 139 that have flowed into the filter unit 1 from the flow path 4 move upward and remain in the space 140 formed in the upper part of the filter unit 1. And the bubble of the space 140 in the filter unit 1 can be discharged outside by opening the valve 142 attached to the upper part. Thus, by providing a deaeration means, the air mixed in the flow path 4 can be removed, and circulating cancer cells can be captured and measured thereafter with higher accuracy.

  Next, in the modification shown in FIG. 17, in addition to the second degassing flow path shown in FIG. 16, a configuration for vibrating the filter unit 1 with a piezoelectric element is added. In this configuration, the filter unit 1 is fixed on the minute vibration generating stage 144 using the piezoelectric element 145, and vertical vibration as indicated by a curve 146 is applied to the filter unit 1 by the piezoelectric element 145. The vibration causes the air bubbles in the filter unit 1 to be guided upward, and discharged to the outside through the pipe 141. That is, according to the configuration shown in FIG. 16, the bubbles are vibrated by the momentum by the piezoelectric element 145 to facilitate the discharge to the filter unit 1.

  Next, in the modification shown in FIG. 18, in addition to the configuration shown in FIG. 14, the gas stored in the container 132 and the gas in the flow path using the valve 151 (flow path valve) provided on the flow path are used. In particular, a configuration for discharging the air flowing into the filter unit 1 will be described. In the configuration shown in FIG. 18, a container 132 containing a fluid for discharging bubbles is attached to the flow path 4, and the pump 14 is driven after switching between the valve 149 and the valve 151, thereby This liquid can be guided to the filter unit 1 and the bubbles in the filter unit 1 can be discharged toward the pump 33. At this time, by increasing the flow rate by the pump 33, the momentum can be increased and the discharge effect can be enhanced. By this operation, the bubbles in the filter unit 1 can be effectively discharged.

(About change of liquid feeding means)
Finally, the configuration of the flow path using the decompression pump will be described with reference to FIG. In the configuration shown in FIG. 19, a flow meter 125 is attached to the tube 124 on the downstream side of the filter unit 1, and this can measure the blood flow rate in the tube 124. In addition, a decompression tank 126 is connected to the downstream side of the tube 124 so that the vacuum generator 127 can control the pressure in the decompression tank 126. That is, when the pressure in the decompression tank 126 is lowered, the flow rate of the compressed air supplied through the compressed air supply pipe 129 is increased by the regulator 128 that varies the pressure of the compressed air, and the suction capacity of the vacuum generator 127 is increased. The pressure in the decompression tank 126 can be measured with a pressure gauge 130. When capturing circulating cancer cells in blood using this device, for example, by capturing the circulating cancer cells, if the resistance of the filter 57 is increased, the flow rate flowing through the filter 57 is secured, In order to maintain the capture capability, the blood flow rate in the tube 124 is measured by the flow meter 125, and the pressure is adjusted by the decompression tank 126 so that this value is constant, thereby capturing the circulating cancer cells in the filter 57. Can be performed with higher accuracy.

  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 deaeration means is not limited to the above-described configuration, and may have other configurations. In the above-described embodiment, the configuration in which the deaeration unit is provided in the blood channel has been described. However, in the case where the filter unit 1 is not provided with the deaeration unit, for example, a configuration in which a deaeration unit is separately provided in the processing liquid channel. Also good.

DESCRIPTION OF SYMBOLS 1 ... Filter unit, 3, 4 ... Channel, 5 ... Treatment liquid supply container, 10 ... Blood supply container.

Claims (22)

A blood storage container for storing blood;
An introduction flow path for introducing a treatment liquid for treating the blood or circulating cancer cells in the blood, a discharge flow path for discharging the blood or the treatment liquid to the outside, and a plurality of flow paths A filter having a through-hole formed in a thickness direction and disposed on a flow path between the introduction flow path and the discharge flow path so that the blood or the treatment liquid passes through the through-hole. With units;
A blood channel that connects the front Kishirube Iriryu path of the filter unit and the blood storage container;
A processing liquid storage container for storing the processing liquid for processing circulating cancer cells captured by the filter by passing through the filter;
A treatment liquid flow path connecting the treatment liquid storage container and the introduction flow path of the filter unit;
Selecting means for selecting a liquid to be supplied to the filter unit from the blood and the treatment liquid;
Liquid feeding means for supplying blood from the blood storage container or processing liquid from the processing liquid storage container to the filter unit based on a selection result of the selection means;
Deaeration means for capturing air in the blood or the treatment liquid and discharging it to the outside upstream of the filter;
A circulating cancer cell capturing apparatus comprising: The deaeration means includes an air reservoir provided on the blood flow path or the treatment liquid flow path,
The upstream side of the air reservoir in the blood channel or the processing liquid channel supplies the blood or the processing liquid from the upper part of the air reservoir to the inside of the air reservoir,
The circulating cancer cell capturing apparatus according to claim 1, wherein the blood or the treatment liquid is discharged from a lower portion of the air reservoir downstream of the air reservoir in the blood channel or the treatment liquid channel.   The deaeration means is inclined so that one end thereof is connected to the blood flow path or the processing liquid flow path, and the position gradually becomes higher toward the other end. The circulating cancer cell capturing device according to claim 1, further comprising a first deaeration channel, wherein the other end is an open end. The deaeration means includes
A storage container connected to the other end of the first degassing channel and storing blood from the first degassing channel;
A first flow path that is provided on the blood flow path or the processing liquid flow path and upstream of the connection position with the first degassing flow path to open and close the blood flow path or the processing liquid flow path. A valve,
With
The circulating cancer cell capturing apparatus according to claim 3, wherein the liquid feeding means is driven in a state where the first flow path valve is closed to feed blood stored in the storage container to the filter unit.   The circulating cancer cell capturing apparatus according to claim 1, wherein the degassing unit includes a second degassing flow path that branches downstream from the introduction flow path of the filter unit and extends upward.   The circulating cancer cell capturing apparatus according to claim 4, wherein the deaeration means further includes a vibrator using a piezoelectric element installed so as to be in contact with the filter unit. A plurality of the treatment liquid storage containers are provided,
The processing liquid flow path includes a plurality of upstream flow paths to which a first end that is one end of each of the plurality of processing liquid storage containers is connected, and the plurality of upstream flow paths. A downstream channel that connects one of the second ends, which is an end different from the first end, and the introduction channel of the filter unit; and the second of the plurality of upstream channels. A selection valve that can be connected to the downstream flow path by selecting any one of the end portions of
The circulating cancer cell capture device according to any one of claims 1 to 6, wherein the liquid feeding unit controls the selection valve based on a selection result of the selection unit. A plurality of the treatment liquid storage containers are provided,
The processing liquid flow path includes a suction nozzle that sucks the processing liquid in the processing liquid storage container, a nozzle tube that supplies the processing liquid sucked by the suction nozzle to the introduction flow path of the filter unit, Position change means for changing the position of the suction nozzle with respect to the processing liquid storage container,
2. The liquid feeding unit performs liquid feeding by moving the suction nozzle by the position changing unit based on a selection result of the selection unit and sucking the processing liquid in the processing liquid storage container. The circulating cancer cell capture device according to any one of -6.   The circulating cancer cell capturing apparatus according to claim 8, wherein the position changing unit changes the position of the suction nozzle with respect to the processing liquid storage container by moving the processing liquid storage container.   The circulating cancer cell capturing apparatus according to claim 8 or 9, wherein the position changing means changes the position of the suction nozzle relative to the processing liquid storage container by moving the suction nozzle.   The circulating cancer cell capture according to any one of claims 8 to 10, wherein the change of the position of the suction nozzle with respect to the treatment liquid storage container is performed using a rectilinear moving table that moves only along a predetermined direction. apparatus.   12. The apparatus according to claim 8, further comprising at least one of a cleaning mechanism that cleans an area of the suction nozzle that contacts the processing liquid stored in the processing liquid storage container and an exchange mechanism that replaces the area. The circulating cancer cell capturing device according to Item. A plurality of the filter units are provided,
Corresponding to the plurality of filter units, the blood flow path and the treatment liquid flow path are respectively provided,
The selection unit selects a liquid to be supplied to the plurality of filter units, and the liquid feeding unit selects from the blood from the blood storage container or the processing liquid storage container based on the selection result of the selection unit. The circulating cancer cell capture device according to any one of claims 1 to 6, wherein the treatment liquid is supplied to each of the plurality of filter units.   The circulating cancer cell capturing apparatus according to claim 13, wherein a plurality of the liquid feeding means are provided so as to correspond to the plurality of filter units.   The circulating cancer cell capturing device according to any one of claims 1 to 14, further comprising circulating cancer cell specifying means for identifying and counting the number of circulating cancer cells captured on the filter. As the circulating cancer cell identification means,
Comprising image acquisition means for acquiring a circulating cancer cell image on the filter;
The circulating cancer cell capturing device according to claim 15, wherein the circulating cancer cell is identified and counted based on an image acquired by the image acquiring means. As the circulating cancer cell identification means,
The circulating cancer cell capturing apparatus according to claim 16, further comprising output means for outputting an image acquired by the image acquisition means.   The circulating cancer cell capturing device according to claim 13, further comprising circulating cancer cell specifying means for identifying and counting the number of circulating cancer cells captured on the filter corresponding to each of the plurality of filter units.   The circulating cancer cell capturing apparatus according to claim 13 or 18, wherein the plurality of filter units are integrated. The circulating cancer cell capturing apparatus according to any one of claims 1 to 19, wherein the liquid feeding unit includes a liquid feeding pump.   21. The circulating cancer cell capturing device according to claim 20, wherein the liquid feeding pump is a peristaltic pump. The liquid feeding means is configured to include a pressure-variable decompression tank connected to the flow path and a flow meter for measuring the pressure in the flow path, and according to the measured value of the flow meter, The circulating cancer cell capture device according to any one of claims 1 to 19 , wherein the tank pressure is changed.

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