JP2015097499A - Cell capture system, and operating method of cell capture system - Google Patents

Abstract

PROBLEM TO BE SOLVED: To make it possible to detect abnormality in a cell capture device or its neighborhood and accurately perform processing related to the capture of a cell.SOLUTION: According to a cell capture system 100 and an operating method thereof, a pressure gauge 21 measures a fluid pressure of a test solution or a processing liquid that flows in a flow channel 9 (a flow channel for discharge out of a system), and based on this fluid pressure, a control unit 30 which is an abnormality determination means, determines whether the upstream side is in an abnormal operation state by using the pressure gauge 21. Thereby, in processing related to the capture of the cell, when any abnormality has occurred in a cell capture device 1 or its neighborhood, it is possible to know as a system that the processing is performed in abnormal operation state. Since the system can deal with, for example, stopping of the processing to recover from the abnormal operating state, or the like, the system prevents the processing related to the capture of the cell in abnormal operating state from continuing, and it becomes possible to efficiently and accurately perform the processing related to the capture of the cell.

Description

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

  Cancers called “malignant tumors” have serious problems in maintaining the life 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 cancer cells moving to other organs. Therefore, the presence or absence and the amount of cancer cells in the blood is important information for predicting cancer metastasis. Cancer cells that circulate in a 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 anti-cancer evaluation, 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. 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 immune cells is subjected to physical action, chemical action and physiological activity. A body fluid treatment system including a cell treatment cartridge capable of providing at least one kind of 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

  Here, when the devices described in Patent Documents 1 to 4 are used, if a narrowing of a flow path or clogging of a filter occurs, it is necessary to recover them. However, it is difficult to visually monitor a series of processes by the cell trapping system.

  The present invention has been made in view of the above, a cell capturing system capable of detecting abnormalities in a cell capturing device and its vicinity, and performing processing related to cell capturing with high accuracy, and a method for operating the cell capturing system. The issue is to provide.

  In order to achieve the above object, a cell capture system 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 liquid or a filter disposed so that the treatment liquid passes through the through-hole, and a selection for selecting a liquid to be supplied to the cell capture device from the test liquid and the treatment liquid And a liquid supply means for supplying the test liquid or the treatment liquid to the cell trapping device based on the selection result of the selection means, and a discharge passage connected to the discharge passage. The test solution or the treatment solution to be discharged is discharged to the outside. An out-of-system discharge channel, a hydraulic pressure measuring unit that measures the liquid pressure of the test liquid or the treatment liquid flowing in the out-of-system discharge channel, and a hydraulic pressure measured by the hydraulic pressure measuring unit And an abnormality determining means for determining whether or not the upstream side of the hydraulic pressure measuring means is in an abnormal operation state.

  Further, the operation method of the cell capturing system 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 test A discharge flow path for discharging the liquid or the treatment liquid to the outside, and a plurality of through-holes are formed in the thickness direction, and the test liquid or the test liquid is placed on the flow path between the introduction flow path and the discharge flow path A filter that is arranged so that the treatment liquid passes through the through-hole, and a selection unit that selects 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, the liquid supply means for supplying the test liquid or the treatment liquid to the cell trapping device and the discharge flow path connected to the discharge flow path System for discharging the test liquid or the processing liquid to the outside And a fluid pressure measuring means for measuring a fluid pressure of the test liquid or the treatment liquid flowing in the extravasation fluid passage by a fluid pressure measuring means. And an abnormality determination step of determining whether or not the upstream side of the hydraulic pressure measurement unit is in an abnormal operation state based on the hydraulic pressure measured in the hydraulic pressure measurement step. Features.

  According to the cell capturing system and the method for operating the cell capturing system described above, the fluid pressure measuring means measures the fluid pressure of the test liquid or the treatment liquid flowing in the flow path for discharge outside the system. Based on this, the abnormality determining means determines whether or not the upstream side of the hydraulic pressure measuring means is in an abnormal operation state. As a result, when any abnormality occurs in the cell capturing device or in the vicinity thereof in the processing related to the cell capturing, the system can grasp that the processing has been performed in the abnormal operation state, for example, stop the processing. Therefore, it is possible to cope with the recovery of the abnormal state and the like, and it is possible to efficiently and accurately perform the process related to the cell capture by preventing the process related to the cell capture in the abnormal operation state from continuing.

  Here, the abnormality determining means holds in advance information serving as a hydraulic pressure threshold for determining the presence or absence of abnormal operation, and compares the hydraulic pressure measured by the hydraulic pressure measuring means with the threshold. Thus, it is preferable to determine whether or not it is an abnormal operation state.

  As described above, the abnormal pressure based on the measured hydraulic pressure is determined by comparing the hydraulic pressure measured by the hydraulic pressure measuring means with the previously held threshold value to determine whether or not it is in an abnormal operation state. It is possible to quickly determine whether or not there is an operation.

  In addition, the abnormality determination unit previously stores target hydraulic pressure information that is information indicating a target range of the hydraulic pressure with respect to an elapsed time from the start of supply of the test liquid or the processing liquid by the liquid feeding unit during normal operation. Holding and comparing the hydraulic pressure measured by the hydraulic pressure measuring means at a predetermined elapsed time from the predetermined time point with the target hydraulic pressure information at the elapsed time to determine whether or not an abnormal operation state has occurred. It is preferable to adopt a mode for determining whether or not.

  As described above, whether the fluid pressure measured by the fluid pressure measuring unit at a predetermined elapsed time from the predetermined time point is compared with the target fluid pressure information at the elapsed time, thereby being in an abnormal operation state. In the case of determining whether or not, for example, even when the hydraulic pressure threshold fluctuates depending on the elapsed time, it can be accurately determined whether or not it is in an abnormal operation state according to the elapsed time.

  Further, the target hydraulic pressure information is a liquid for an elapsed time from a predetermined time when each of the plurality of types of liquids is supplied based on a supply order of the plurality of types of liquids supplied to the cell capturing device. It is information indicating the target range of pressure, and it is preferable that the predetermined time point is created as a time point before the start of supply of the first type of liquid.

  As described above, when the plurality of types of liquids supplied to the cell trapping device are supplied based on the supply order from the time before the supply of the first type of liquid is started, Information indicating the target range is held as target hydraulic pressure information, and based on this, it is determined whether or not it is in an abnormal operation state. Since it is possible to integrally determine whether or not it is in an operating state, it is possible to accurately and efficiently determine whether or not it is in an abnormal operating state.

  Further, the apparatus further comprises a flow rate measuring means for measuring the flow rate of the test liquid or the treatment liquid flowing in the out-of-system discharge flow path, and the abnormality determining means is based on the flow rate measured by the flow rate measuring means. It is preferable that the upstream side of the hydraulic pressure measuring unit and the flow rate measuring unit determine whether or not the abnormal operation state is present.

  As described above, by determining whether or not it is in an abnormal operation state based on the flow rate measured by the flow rate measuring means, the determination as to whether or not it is in an abnormal operation state is based on two pieces of information. Since it can be performed, it can determine more accurately.

  ADVANTAGE OF THE INVENTION According to this invention, the operation method of the cell capture system which can detect the abnormality in a cell capture device or its vicinity, and can perform the process which concerns on the capture | acquisition of a cell accurately is provided.

It is a figure explaining the structure of the cell capture system which concerns on embodiment of this invention. It is a figure explaining the structure of the cell capture device which comprises a cell capture system. It is a figure explaining the driving | running state monitoring method in a cell trapping system. It is a figure explaining an example of the driving | running state monitoring method. It is a figure explaining other examples of a driving state monitoring method. It is a figure explaining the usage method of the flowmeter of a driving | running state monitoring method.

  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 system)
FIG. 1 is a diagram illustrating a configuration of a cell trapping system according to this embodiment. The cell capture system is a device that captures cells contained in a test liquid by filtering a cell dispersion liquid to be the 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 trapping system, 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 system 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. 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 a sealing 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.

  Connected to the flow path 4 connected to the cell capture device 1 is a test liquid storage container 10 in which blood containing cells is stored as a test liquid. 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.

  Further, when supplying any one of the treatment liquid and the test liquid to the cell trapping device 1, a flow path 9 (external discharge flow path) made of a soft tube downstream of the cell trapping device 1. The liquid is supplied by sucking the target liquid by driving a pump 14 (supply means) provided above. 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.

  A pressure gauge 21 (hydraulic pressure measuring means) and a flow meter 23 (flow rate measuring means) are connected to the flow path 9 on the downstream side of the cell trapping device 1. In FIG. 1, the pressure gauge 21 is connected to the upstream side of the pump 14. The pressure gauge 21 is also connected to the upstream side of the cell trapping device 1 and to the downstream side of the valve 12 on the flow path 3 connecting the processing liquid storage container 5 and the cell trapping device 1. Thereby, the pressure gauge 21 is provided so that the liquid pressure of the liquid flowing through the flow path 9 and the liquid pressure of the liquid flowing through the flow path 3 can be measured. In the cell trapping system 100 according to the present embodiment, the liquid pressure of the liquid flowing through the flow path 3 and the flow path 9 are set as the liquid pressure of the liquid flowing through the flow path 9 when the treatment liquid is flowed to the cell trapping device 1. The pressure difference (differential pressure) with the liquid pressure of the liquid flowing through is calculated and used. In addition, the liquid pressure of the liquid flowing through the flow path 9 when the test liquid is flowed to the cell capture device 1 is replaced with a configuration in which the liquid pressure of the test liquid on the upstream side of the cell capture device 1 is measured. The atmospheric pressure is used, and the pressure difference (differential pressure) between the atmospheric pressure and the liquid pressure of the liquid flowing through the flow path 9 is calculated and used. In order to measure the fluid pressure of the test fluid upstream of the cell capture device 1, the fluid pressure is measured in the flow path 4 connecting the test fluid storage container 10 and the cell capture device 1 to measure the differential pressure. However, in that case, it is necessary to replace the pressure gauge every time a new test solution is measured (to replace the test solution) in order to prevent foreign matter from entering. Note that the configuration for calculating the differential pressure as in the present embodiment is not essential, and the measurement position by the pressure gauge 21 may be configured to include only the flow path 9 on the downstream side of the cell trapping device 1.

  The flow meter 23 is provided on the downstream side of the pump 14 and measures the flow rate of the liquid flowing through the flow path 9. Information on the pressure and flow rate of the liquid measured by the pressure gauge 21 and the flow meter 23 is used to confirm whether the cell capture process by the cell capture system 100 is appropriately performed. This point will be described later. The measurement positions of the pressure gauge 21 and the flow meter 23 in the flow path 9 are not particularly limited as long as they are downstream of the cell capture device 1. When the hydraulic pressure is measured on the upstream side of the cell capture device 1 as in this embodiment, the measurement position is not particularly limited as long as the measurement position is on the flow path upstream of the cell capture device 1.

  The above-described control of each unit is performed by control by the control unit 30 (selection unit, liquid feeding unit, abnormality determination unit). Specifically, the selection valve 8, the valves 12 and 13, and the pump 14 are driven by an instruction from the control unit 30. The control unit 30 is provided with a program input function for inputting a program that enables control such as driving and stopping for each of the above-described units. A drive mechanism that operates in order is added. The control unit 30 selects a line through which the liquid flows, and the control unit 30 instructs each unit to open and close the valve and drive the pump based on the selection result.

  Further, the control unit 30 has a function of evaluating whether or not the cell trapping system 100 is in an abnormal operation state based on information from the pressure gauge 21 and the flow meter 23. And when it is evaluated that there is some abnormality in the cell capture system 100, while recording about an abnormal operation state, it can also be set as the structure which stops operation | movement of the cell capture system 100 depending on the case. The operation state evaluation by the control unit 30 and the control based on the evaluation result will be described later.

  Next, the cell trapping device 1 will be described with reference to FIG. 2A is a top view of the cell trapping device 1, and FIG. 2B is a cross-sectional view taken along the line IIB-IIB in FIG. 2A.

  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.

  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. The “treatment liquid flow path” and the “test liquid flow path” refer to the flow paths 3 and 4 respectively connected to the upstream sides of the flow paths 3A and 4A of the cell trapping device 1.

  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 cell 65 that is a capture target from passing.

(Operation method of cell capture system)
In the cell capturing system 100 having the above-described configuration, first, the test liquid in the test liquid storage container 10 is introduced into the cell capturing device 1 through the flow path 4. In the cell capture device 1, the test liquid is introduced from the flow path 4 A for the test liquid, passes through the filter 57, is discharged from the flow path 9 A, and is sent to the waste liquid container 16. At this time, since the cell that is the capture target cannot pass through the through hole 61 of the filter 57, the cell 65 is captured on the filter 57.

  Next, washing and staining are performed to detect the captured cells. Washing and staining are performed by supplying a washing solution, a fixing solution, a permeating solution, and a staining solution from the channel 3 of the tube connected to the channel 3A after filtering of the test solution using the cell capture device 1 is completed. Done. By this operation, cell capture using the filter 57 and staining with the treatment liquid are performed. Moreover, the cell capture device 1 is removed from the flow paths 3 and 4 and the flow path 9 for identification of cells and measurement of the number of cells as required.

  These operations are controlled by the control unit 30 so that capturing of cells in the test solution by the filter 57 of the cell capturing device 1 and washing and staining of the captured cells are performed as a series of operations related to capturing the cells. Done.

  Here, the monitoring method of the driving | running state by the control part 30 is demonstrated using FIG. FIG. 3 is a flowchart for explaining an operation state monitoring method by the control unit 30.

  First, the principle of operation state monitoring will be described. As described above, the control unit 30 of the cell trapping system 100 has a function of evaluating the operating state of the cell trapping system 100, that is, whether it is a normal operation state or an abnormal operation state. Here, the abnormal operation means that the fluid in the flow paths 3, 4, 9 and the cell trapping device 1 does not flow properly as expected for some reason. The test liquid and the processing liquid are introduced into the cell trapping device 1 from the test liquid storage container 10 or the processing liquid storage container 5 by driving the pump 14, and pass through the filter 57 in the cell trapping device 1. The cells to be captured are captured by the filter 57, and the liquid containing the other cells is discharged to the waste liquid container 16 through the discharge channel 9A and the channel 9. Such abnormalities in the cell trapping system 100 may be, for example, constriction in any flow path, clogging of the filter 57, or the like. When such a phenomenon occurs, even if the rotation speed of the pump 14 is constant and the liquid in the flow path is sucked at a constant pressure, the liquid in the flow path can be sufficiently sucked by the pump 14. As a result, the hydraulic pressure in the pressure gauge 21 increases compared to the normal operation state, and the flow rate in the flow meter 23 decreases compared to the normal operation state.

  Further, for example, it is conceivable that the flow paths 3, 4, 9 are not properly connected to the cell trapping device 1. When such a phenomenon occurs, even if the rotation speed of the pump 14 is constant and the liquid in the flow path is sucked at a constant pressure, the liquid in the flow path can be sufficiently sucked by the pump 14. As a result, the hydraulic pressure in the pressure gauge 21 decreases as compared with the normal operation state, and the flow rate in the flow meter 23 decreases as compared with the normal operation state.

  Therefore, in the cell trapping system 100, when any problem occurs in the upstream channel, the cell trapping system 100 is utilized by utilizing the fact that the fluid pressure and flow rate in the channel greatly change. The configuration of monitoring the operating state of the was adopted.

  Returning to FIG. 3, as a method of monitoring the operation state by the control unit 30, first, the fluid pressure and the flow rate are acquired from the pressure gauge 21 and the flow meter 23 as information (evaluation information) for evaluating the operation state ( S10, hydraulic pressure measurement step). Information related to the hydraulic pressure measured by the pressure gauge 21 and information related to the flow rate measured by the flow meter 23 are sent from the pressure gauge 21 and the flow meter 23 to the control unit 30.

  In the control part 30, based on the information sent from the pressure gauge 21 and the flowmeter 23, evaluation about an operation state is performed (S20, abnormality determination step). Then, it is determined whether or not it is in an abnormal operation state (S30, abnormality determination step). As a result, when it is determined that the operation state is abnormal, the control unit 30 operates the cell capture system 100. Is stopped (S40). Further, when it is determined that the state is not an abnormal operation state (a normal operation state), the control unit 30 ends the process related to the evaluation. By repeating such processing, it is possible to evaluate the operation state of the cell trapping system 100 in real time. Note that the process shown in FIG. 3 in which the operation is stopped by the control unit 30 when it is determined to be in an abnormal operation state (S40) is an example of the process in the case where it is determined to be in an abnormal operation state, It can be changed as appropriate.

(Abnormal operation status judgment method)
Next, a method used when the control unit 30 evaluates the operating state based on information sent from the pressure gauge 21 and the flow meter 23 (S20, S30) will be described. Broadly speaking, there are (1) a method using a threshold value and (2) a method using target hydraulic pressure information during normal operation. In the following description, a determination method based on information on the hydraulic pressure from the pressure gauge 21 that is an essential configuration in the cell trapping system according to the present embodiment will be described, and then information on the flow rate from the flow meter 23 will be described. The determination method when using also will be described.

(1-Method using threshold)
First, the method using the threshold is that the information related to the hydraulic pressure threshold for determining the presence or absence of abnormal operation is stored in the control unit 30 in advance, and the hydraulic pressure measured by the pressure gauge 21 and the control unit 30 This is a method for determining whether or not an abnormal operation state exists by comparing with a held threshold value. For example, when the upper limit value and the lower limit value of the allowable range of the hydraulic pressure measured in the pressure gauge 21 in the normal operation state are set as threshold values, the hydraulic pressure in the pressure gauge 21 is higher than the upper limit side threshold value, or This is a method of determining that the cell trapping system 100 is in an abnormal operation state when it is lower than the lower limit side threshold. By using this method, it is possible to easily determine that the operation state is abnormal.

  However, in the cell trapping system 100, a test solution and a treatment solution having different viscosities are applied to cells according to the progress of the series of operations during a series of operations for performing a process of capturing cells contained in one test solution. It is necessary to flow toward the capture device 1. It is conceivable that the inflow speed and the like at this time need to be changed depending on the liquid introduced into the cell trapping device 1. Moreover, in the time slot | zone which stops introduction | transduction of a liquid with respect to the cell capture device 1, a liquid pressure or a flow volume may consider removing from a threshold value. Therefore, it is conceivable to use (2) a method that utilizes target hydraulic pressure information during normal operation.

(2-Method of using target hydraulic pressure information during normal operation)
“Target hydraulic pressure information” in the method of using the target hydraulic pressure information during normal operation is information indicating a target range of hydraulic pressure with respect to an elapsed time from a predetermined time point. FIG. 4 illustrates the case where the blood that is the test liquid passes through the filter 57. As a predetermined time point, here, the elapsed time from the time when the liquid supply of the test liquid is started is shown on the horizontal axis. When the pressure is set and the pressure (fluid pressure) is set on the vertical axis, the fluctuation of the liquid pressure assumed during normal operation is shown as an assumed fluctuation C0. In FIG. 4, it is recorded that the fluid pressure increases at the start time T1 and decreases at the end time T2 by flowing the test solution. This is because the liquid pressure increases because components such as red blood cells contained in blood pass through the filter 57, for example, when the blood has passed through the filter 57 by flowing the next liquid (for example, washing liquid) (T2). ) Shows that the hydraulic pressure decreases. At this time, an allowable range is provided on the low pressure side and the high pressure side with respect to the assumed fluid pressure fluctuation C0 during normal operation, and the low pressure side lower limit fluctuation C1 and the high pressure side upper limit fluctuation C2 are set. And between the lower limit fluctuation | variation C1 and the upper limit fluctuation | variation C2 with respect to predetermined | prescribed elapsed time becomes the target range of the hydraulic pressure with respect to the elapsed time from a predetermined | prescribed time.

  When the target range is set as described above, if the hydraulic pressure measured by the pressure gauge 21 at the elapsed time t from a predetermined time is P, the control unit 30 refers to FIG. Based on whether P is included between the value of the lower limit variation C1 and the value of the upper limit variation C2 at time t, a determination is made as to whether or not it is in a normal operating state. And when it determines with it being a normal operation state, while continuing the process which concerns on the to-be-tested liquid using the cell capture system 100, the measurement by the pressure gauge 21 and the determination of the normal operation state by the control part 30 are carried out. repeat. Thereby, when a series of processes related to the test liquid using the cell trapping system 100 is performed in a normal operation state, that is, when abnormality is monitored and it is determined that the operation is abnormal, the operation is performed. Can be stopped.

  In addition, in FIG. 4, the target fluid pressure information related to the change in fluid pressure when blood as the test solution passes through the filter 57 has been described, but in actuality, in a series of processes for capturing cells of the test solution. Needs to process the captured cells by allowing a plurality of types of processing liquids to pass through the filter 57 in addition to the test liquid. Therefore, it is preferable to collectively monitor the pressure fluctuation when the test solution and the plurality of processing solutions are sequentially passed through the filter 57.

  As a method for solving the above-described object, the target hydraulic pressure information held in the control unit 30 supplies a plurality of types of liquids based on the supply order of the plurality of types of liquids supplied to the cell trapping device 1, respectively. There is a method of using information created as information indicating the target range of the hydraulic pressure with respect to the elapsed time from the time before the start of supplying the first type of liquid. This point will be further described with reference to FIG.

  FIG. 5 shows fluctuations in fluid pressure when a plurality of types of liquids, that is, a test solution and a plurality of types of processing solutions are sequentially supplied to the cell trapping device 1. Specifically, the time point (start of driving of the pump 14) at the start of feeding of the test solution in the cell trapping system 100 is set as the starting point (time 0), the cell trapping process (supply of the test solution) L1, and the cleaning process ( Cleaning liquid supply) L2, fixing process (fixed liquid supply) L3, permeation process (permeate supply) L4, staining process (cleaning liquid supply) L5, and cleaning process (cleaning liquid supply) L6. It shows the fluctuation of the corresponding hydraulic pressure. In FIG. 5, regarding the cell capture process L1, the pressure difference (differential pressure) between the atmospheric pressure and the liquid pressure of the liquid flowing through the flow path 9 is shown. For the processes after the cleaning process L2, the liquid flowing through the flow path 3 is shown. A pressure difference (differential pressure) between the liquid pressure and the liquid pressure of the liquid flowing through the flow path 9 is shown.

  Here, assuming that the hydraulic pressure fluctuation shown in FIG. 5 corresponds to the assumed fluctuation C0 during normal operation in FIG. 4, the hydraulic pressure during normal operation so as to correspond to the lower limit fluctuation C1 and the upper limit fluctuation C2 shown in FIG. If an allowable range is provided on the low pressure side and the high pressure side with respect to the assumed fluctuation C0, information corresponding to the lower limit fluctuation C1 on the low pressure side and the upper limit fluctuation C2 on the high pressure side shown in FIG. 4 is obtained. The information corresponding to the lower limit fluctuation C1 and the upper limit fluctuation C2 obtained in this way can be used as target hydraulic pressure information relating to a series of processes relating to cell capture. Thereby, in the control part 30, the hydraulic pressure measured by the pressure gauge 21 according to the elapsed time from the predetermined time point (here, the drive start time of the pump 14) is between the lower limit fluctuation value and the upper limit fluctuation value. Is determined based on whether it is included in the normal operation state.

  By adopting such a configuration, for example, in the fixing process L3, when the fluid pressure becomes 0 due to disconnection somewhere in the upstream flow path, abnormal operation is performed in the control unit 30. It can be determined as a state, and a series of operations related to cell capture can be stopped. In this way, at any point in the series of operations related to cell capture, monitoring according to the progress of the operation can be performed, and when an abnormality occurs, an appropriate response can be made.

(3-Use of information from the flow meter)
In the above, the method for detecting an abnormal operation state in the cell trapping system 100 based on the fluid pressure measured by the pressure gauge 21 has been described. However, in the cell trapping system 100 according to the present embodiment, measurement is performed by the pressure gauge 21. In addition to the hydraulic pressure, information related to the flow rate measured by the flow meter 23 can also be used for detecting an abnormal operation state.

  As a specific method of use, instead of the above-described (1-method using a threshold) and (2-method using target hydraulic pressure information during normal operation), the evaluation target is a hydraulic pressure. What is necessary is just to make flow volume into evaluation object. That is, (1) information related to a flow rate threshold value for determining the presence or absence of abnormal operation is held in the control unit 30 in advance, and the flow rate measured by the flow meter 23 is compared with the threshold value held in the control unit 30 And (2) target flow rate information during normal operation (information indicating a target range of flow rate with respect to an elapsed time from a predetermined time point) This is a method for determining whether or not an abnormal operation state is present depending on whether or not the flow rate measured by the flow meter 23 is included between the lower limit fluctuation and the upper limit fluctuation.

  FIG. 6 is a diagram illustrating an example of flow rate measurement data measured by the flow meter 23. In FIG. 6, the horizontal axis indicates the elapsed time from a predetermined time point, and the vertical axis indicates the flow rate of the liquid flowing through the flow path 9. FIG. 6 shows the progress from the start to the end of a series of operations related to cell capture. For example, at time T5, the liquid is flowed at a flow rate of 200 μL / min (passes through the installation position of the flow meter 23 in the flow path 9), and then stops for 20 minutes, and then again at a flow rate of 200 μL / min from time T6. It is shown that the liquid has flowed. Thereafter, the state where the flow of the liquid is stopped and the state where the liquid is flowed at the determined flow rate continue alternately, indicating that the movement of the liquid is completed at time T7 of 106 minutes after the start. By comparing the flow rate information relating to the movement of the liquid in FIG. 6 and the target flow rate information during normal operation, the control unit 30 can determine whether or not it is in an abnormal operation state.

  Further, in the cell trapping system 100 according to the present embodiment, the information of the fluid pressure by the pressure gauge 21 and the information of the flow rate by the flow meter 23 are combined to control the status of each device upstream of these instruments. The part 30 can be grasped more accurately. Specifically, it can be used for monitoring a cell capture state of a filter of a cell capture device for capturing cells. In other words, if the pressure drops and the flow rate decreases by more than a certain slope with respect to time, it turns out that the filter is clogged for some reason, and as a monitor that performs operations to eliminate this Can be used. Moreover, since the flowmeter 23 can be used also for the measurement or control of the exact flow volume which concerns on a test liquid, it can improve the precision of a measurement result.

  In addition, the information on the fluid pressure measured by the pressure gauge 21 and the information on the flow rate measured by the flow meter 23 are, for example, information specifying the test liquid and information specifying the work execution date and time in the control unit 30. By holding the data in association with each other, it can also be used as log data indicating that a series of operations related to the capture of cells in the test solution have been performed appropriately. It can also be used as one piece of information that enhances the reliability.

  As described above, according to the cell trapping system 100 and the operation method thereof according to the present embodiment, the pressure of the test liquid or the processing liquid flowing in the flow path 9 (external discharge flow path) is measured by the pressure gauge 21. Is measured, and based on this hydraulic pressure, it is determined by the control unit 30 which is an abnormality determination means whether or not the upstream side of the pressure gauge 21 is in an abnormal operation state. As a result, when any abnormality occurs in the cell capturing device 1 or in the vicinity thereof in the processing related to the cell capturing, the system can grasp that the processing has been performed in the abnormal operation state, for example, stop the processing. Therefore, it is possible to cope with the recovery of the abnormal state and the like, and it is possible to efficiently and accurately perform the process related to the cell capture by preventing the process related to the cell capture in the abnormal operation state from continuing.

  In particular, the filter 57 of the cell trapping device 1 used in the cell trapping system 100 according to the present embodiment is a metal thin film filter, and the amount of cells to be trapped contained in the test solution like CTC Is a very limited cell, for example, if the internal fluid pressure becomes high due to clogging of the pipe or the like, the filter 57 itself is damaged, and all cells to be captured flow. There is a possibility. Therefore, by adopting a configuration such as the cell trapping system 100 according to the present embodiment, a change in fluid pressure of about ± 2 kPa, for example, compared to a normal operation state before an abnormality that cannot be recovered such as filter breakage occurs. Based on the above, it becomes possible to detect an abnormality.

  As a method for determining whether or not the controller 30 is in an abnormal operation state, there is a method for determining whether or not the fluid pressure measured by the pressure gauge 21 is in an abnormal operation state by comparing the hydraulic pressure with a previously held threshold value. By having such a configuration, it is possible to quickly determine whether there is an abnormal operation based on the measured fluid pressure.

  As another method for determining whether or not the control unit 30 is in an abnormal operation state, the control unit 30 indicates the target range of the fluid pressure with respect to the elapsed time from the start of supply of the test liquid or the processing liquid during normal operation. The target hydraulic pressure information, which is information, is held in advance, and the abnormal pressure is detected by comparing the hydraulic pressure measured by the pressure gauge 21 at a predetermined elapsed time from the start of supply with the target hydraulic pressure information at this elapsed time. It can be determined whether or not. In this case, for example, even when the hydraulic pressure threshold fluctuates depending on the elapsed time, it can be accurately determined whether or not it is in an abnormal operation state according to the elapsed time.

  Furthermore, the target range of the hydraulic pressure with respect to the elapsed time when a plurality of types of liquid supplied to the cell trapping device are supplied based on the supply order from the time before the start of the supply of the first type of liquid. The information shown is held as target fluid pressure information, and based on this, it is determined whether or not it is in an abnormal operation state, and in a series of operations after the start of the supply of the first type of liquid, Since it can be integrally determined whether or not there is an abnormal operation state, it can be accurately and efficiently determined.

  Then, as described above, by determining whether or not it is in an abnormal operation state based on the flow rate measured by the flow meter 23, the determination as to whether or not it is in an abnormal operation state is made into two pieces of information. Since it can be performed based on this, it can be determined more accurately.

  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 system 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 system 100 demonstrated in the said embodiment as needed.

  In addition, the cell capturing system of the present invention and the method of operating this system are useful for capturing cells in blood (red blood cells, white blood cells, platelets, circulating cancer cells in the blood, 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 ... Processing liquid storage container, 10 ... Test liquid storage container, 21 ... Pressure gauge, 23 ... Flow meter, 100 ... Cell capture system.

Claims (6)

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 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 the test liquid or the treatment liquid to the cell capturing device;
An out-of-system discharge flow path that is connected to the discharge flow path and discharges the test liquid or the treatment liquid discharged from the discharge flow path;
A fluid pressure measuring means for measuring a fluid pressure of the test liquid or the treatment liquid flowing in the outside discharge flow path;
An abnormality determining unit that determines whether or not the upstream side of the hydraulic pressure measuring unit is in an abnormal operation state based on the hydraulic pressure measured by the hydraulic pressure measuring unit;
A cell capture system comprising: The abnormality determining means includes
Pre-hold information that serves as a hydraulic pressure threshold for determining the presence or absence of abnormal operation,
The cell trapping system according to claim 1, wherein it is determined whether or not an abnormal operation state is established by comparing the fluid pressure measured by the fluid pressure measuring means with the threshold value. The abnormality determining means includes
During normal operation, target fluid pressure information that is information indicating a target range of fluid pressure with respect to the elapsed time from a predetermined time point is retained in advance,
It is determined whether or not there is an abnormal operation state by comparing the hydraulic pressure measured by the hydraulic pressure measuring means at a predetermined elapsed time from the predetermined time point with the target hydraulic pressure information at the elapsed time. The cell capturing system according to claim 1. The target hydraulic pressure information is a hydraulic pressure with respect to an elapsed time from a predetermined time point when each of the plurality of types of liquids is supplied based on a supply order of the plurality of types of liquids supplied to the cell trapping device. Information indicating the target range,
The cell capture system according to claim 3, wherein the predetermined time point is created as a time point before the start of supplying the first type of liquid. A flow rate measuring means for measuring a flow rate of the test liquid or the processing liquid flowing in the outside discharge flow path;
The abnormality determination unit determines whether the upstream side of the fluid pressure measurement unit and the flow rate measurement unit is in an abnormal operation state based on the flow rate measured by the flow rate measurement unit. The cell trapping system according to claim 1. 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 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 the test liquid or the treatment liquid to the cell capturing device;
An external discharge flow path connected to the discharge flow path to discharge the test liquid or the treatment liquid discharged from the discharge flow path to the outside,
A liquid pressure measuring step for measuring a liquid pressure of the test liquid or the processing liquid flowing in the outside discharge flow path by a liquid pressure measuring means;
Based on the hydraulic pressure measured in the hydraulic pressure measurement step, an abnormality determination step of determining whether the upstream side of the hydraulic pressure measurement means is in an abnormal operation state in the abnormality determination means;
A method for operating a cell trapping system comprising:

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