JP2017070410A - Automatic blood separator - Google Patents

Abstract

PROBLEM TO BE SOLVED: To provide an automatic blood separator for subjecting blood to centrifugation and reliably obtaining prescribed components such as a cell and a lymphocyte.SOLUTION: The automatic blood separator has an enclosure and a controller disposed inside or outside the enclosure, with a centrifugal machine, a sample housing part, an action part and a drive part within the enclosure. The action part has at least a test tube holding mechanism and an auto-pipet mechanism; the drive part transfers the action part in an upper space of the centrifugal machine and the sample housing part; the centrifugal machine has a protective wall surface with a see-through window opened in part of the surrounding lateral face, with an imaging device capable of imaging the inside of the centrifugal machine via the see-through window, disposed outside the see-through window; and the controller controls the centrifugal machine, the action part, the drive part and the imaging device.SELECTED DRAWING: Figure 1

Description

  The present invention relates to an automatic blood separation apparatus that automatically performs a series of operations for separating a specific component from blood.

  As an apparatus for automatically separating and collecting blood components, for example, there is an apparatus disclosed in Patent Document 1. The device of Patent Document 1 is a device for separating necessary components (plasma and platelets) and other components in component blood donation, and has a device main body and a blood collection kit attached to the device main body. The apparatus main body has a box-shaped mechanism main body, the mechanism main body has a centrifuge mechanism, and blood is introduced from the blood collection kit into the centrifuge mechanism, and the introduced blood is centrifuged. Through the device, the desired components are recovered and the remaining components are returned to the donor.

  The blood separation device as described above is suitable for continuously processing a large amount of blood, but for a device that accurately and automatically separates specific cells and lymphocytes from a smaller amount of blood. In such a case, the work is performed manually. Work by hand is generally performed according to the following procedure.

  First, the blood to which the anticoagulant is added is diluted twice with physiological saline, and the specific gravity separation liquid is appropriately dispensed into a round bottom test tube, and the blood dilution liquid is layered on the specific gravity separation liquid so as not to become turbid. . Subsequently, the round-bottom test tube is carried to a centrifuge and set in the centrifuge to adjust the balance as necessary. Subsequently, specific gravity separation is performed at a predetermined number of revolutions, and the test tube is taken out of the centrifuge with extreme care so that the layer separated by specific gravity does not become turbid, and the target layer to be sampled separated by specific gravity is removed from the test tube. Collect with a Pasteur pipette. Next, the collected cells are discharged into a test tube (V-bottom test tube) into which physiological saline has been injected in advance as a washing solution, set in a centrifuge, and separated at a predetermined number of rotations. Then, the test tube is taken out from the centrifuge, and the supernatant is discarded from the test tube to obtain cells and lymphocytes to be collected.

  As described above, the work of separating cells and lymphocytes from blood requires complicated and precise operations and requires a large human burden. Therefore, separate means are required from the viewpoint of technology and cost and from the viewpoint of work accuracy. Was desired.

JP 2007-130283 A

  In view of the above situation, an object of the present invention is to provide an automatic device for subjecting whole blood to centrifugation and reliably obtaining a target component such as cells and lymphocytes.

  The inventor has proceeded with studies to solve the above-described problems, and includes a centrifuge, a sample storage unit, and an operation unit that performs a series of operations on behalf of a human hand, and the control unit performs centrifugation. It was conceived to provide a device capable of automatically performing a series of operations for separating a specific component from a blood sample by controlling the operation of the machine and the operation unit. Further, the inventor has proceeded with the study, provided a transparent window in a part of the protective wall of the centrifuge, arranged the imaging device on the outside of the transparent window, and placed it at a position facing the imaging device across the bucket of the centrifugal rotor. The inventors came up with a configuration in which a light source is arranged. Such a configuration makes it possible to clearly photograph the contents of the centrifuge tube inserted into the bucket, and reliably detects only the desired fraction by detecting the interface position of the blood separated in specific gravity in the centrifuge tube. The present invention was completed.

That is, the present invention is an automatic blood separation device,
A housing and a control unit provided inside or outside the housing;
A centrifuge, a sample storage unit, an operation unit, and a drive unit are provided inside the housing.
The operating unit has at least a test tube holding mechanism and an automatic pipette mechanism,
The drive unit moves the operation unit in an upper space of the centrifuge and the sample storage unit,
The centrifuge has a protective wall surface provided with a see-through window on a part of a peripheral side surface, and an imaging device capable of taking an image of the inside of the centrifuge through the see-through window is provided outside the see-through window. And
The said control part is related with what controls the said centrifuge, the said operation part, the said drive part, and the said imaging device.

  The centrifuge is preferably a swing rotor centrifuge.

  Moreover, it is preferable that a light source is provided inside the centrifuge, and the imaging device and the light source are opposed to each other with a bucket of the centrifuge interposed therebetween.

  In the automatic blood separation device according to the present invention, the casing has a vent, the vent has a HEPA filter, and the centrifuge has a lid surface fixed to the top surface and a bottom surface. The automatic blood separation according to any one of the above, wherein an opening is provided in each of the lid surface and the bottom surface, and only air that has passed through the HEPA filter is vented through the opening in the centrifuge. Relates to the device.

  In the automatic blood separation apparatus of the present invention, it is preferable that the articles are taken into and out of the centrifuge only through the opening on the lid surface of the centrifuge.

  Further, in the automatic blood separation apparatus of the present invention, the operating part includes a test tube holding mechanism that can hold a test tube, an automatic pipette mechanism that attaches a pipette tip to a tip and performs suction and discharge, and a tube connected to a waste liquid bottle. And a suction mechanism that performs only suction.

  According to the present invention, since a series of operations conventionally performed by a person can be automatically executed, human burden is reduced, and desired cells and lymphocytes are reliably obtained from blood without fluctuations in work accuracy. Can be separated. Further, according to the present invention, it is possible to detect the state of the sample in the centrifuge tube (position of the interface) by photographing the test tube after the specific gravity separation with an imaging device provided outside the centrifuge. Only the target component can be reliably collected. Further, by arranging the imaging device outside the protective wall of the centrifuge, there is less risk of contamination and excellent maintainability.

It is a schematic diagram which shows the front inside of the automatic blood separation apparatus of this invention. It is a mimetic diagram showing the inside of the side of the automatic blood separation device of the present invention. It is a schematic diagram which shows the upper surface of the automatic blood separation apparatus of this invention. It is a schematic diagram which shows the inside (during stop) of the centrifuge of the automatic blood separation apparatus of this invention. It is a schematic diagram which shows the inside (during rotation) of the centrifuge of the automatic blood separation apparatus of this invention. It is a schematic diagram which shows the inside (during imaging) of the centrifuge of the automatic blood separation apparatus of this invention. It is a control block diagram which shows the control part of the automatic blood separation apparatus of this invention. It is a flowchart which shows the operation | movement procedure of the automatic blood separation apparatus of this invention. It is a flowchart which shows the operation | movement procedure of the automatic blood separation apparatus of this invention.

  Hereinafter, an automatic blood separation device according to the present invention will be described with reference to FIGS. In this specification, the X-axis direction means the depth direction of the device, the Y-axis direction means the left-right direction of the device, and the Z-axis direction means the up-down direction of the device. Moreover, in each drawing, the same code | symbol is attached | subjected to the same structure and description may be abbreviate | omitted.

  FIG. 1 is a diagram schematically showing the inside of the front of the automatic blood separation device 1 of the present invention. The automatic blood separation device 1 has a housing 2 that covers the entire device, and includes a centrifuge 3, a sample storage unit 4, an operation unit 5, and a driving unit 6 inside the housing 2. The housing 2 is a box-type device exterior partly or entirely made of stainless steel or the like, and a vent 8 that houses a fan and accommodates the HEPA filter 7 is provided on the upper side surface. The outside air taken in from the vent 8 passes through each part inside the housing and is discharged from the exhaust port 9. The centrifuge 3 will be described in detail later.

  The sample storage unit 4 includes a blood collection tube and / or test tube containing blood collected from a patient or a subject, test tubes such as an empty tube for balance adjustment, various liquids such as a specific gravity liquid and a cell washing liquid, various equipment ( A sample rack 10 that can store a disposable pipette tip, a U-bottom test tube, a V-bottom test tube, and the like and can be taken in and out of the housing 2 is housed. The sample rack 10 may be configured to accommodate samples, test tubes, pipette tips, and the like in an integrated rack, or may be composed of a plurality of racks that accommodate equipment and the like by type. In the automatic blood separation apparatus 1 of the present invention, a dedicated disposable tip, a U-bottom test tube, a V-bottom test tube, or the like may be used, or commercially available equipment may be used. The sample rack 10 is placed on the rack rail 11 and can move in the X-axis direction on the rack rail 11 according to the operation of the motor.

  The operation unit 5 includes three operation arms 5a, 5b, and 5c, and the three operation arms are collectively supported by the drive unit 6. The operation unit 5 is supported by the drive unit 6 and can move in the upper space of the centrifuge 3 and the sample storage unit 4. In the example of FIG. 1, the operation unit 5 can move in the X-axis and Y-axis directions. However, the operation unit 5 can be moved only in the Y-axis direction depending on the arrangement of each device constituting the apparatus.

  The drive unit 6 is assembled to the Y-axis rail 12, and the operation unit 5 can move along the Y-axis rail 12 in accordance with the operation of the drive unit 6. The Y-axis rail 12 is attached to the support body 13, and the support body 13 is placed on the X-axis rail 14. That is, the operation unit 5 and the drive unit 6 can move on the X-axis rail while being supported by the support 13. That is, the operation unit 5 can move to any position in the XY plane by a combination of movement in the X-axis direction and movement in the Y-axis direction. Since the three operating arms of the operating unit 5 are collectively supported by the driving unit 6, the three move in the X-axis direction and the Y-axis direction, but move in the Z-axis direction (described later). Then, each arm is controlled independently.

  The operation unit 5 will be described. The operating arm 5a is a test tube holding mechanism, and can fit and hold the test tube at the lower end of the arm and release the holding in accordance with an instruction from the control unit. The test tube holding mechanism 5a moves to a predetermined position, engages the test tube at the lower end, moves with the test tube fitted and held, and performs an operation of releasing the holding at the predetermined position. By this operation, for example, the test tube is conveyed from the sample storage unit 4 to the centrifuge 3 or from the centrifuge 3 to the sample storage unit 4. Further, the position of the test tube can be moved in the sample storage unit 4 (sample rack 10).

  The operating arm 5b is an automatic pipette mechanism, and a disposable pipette tip is fitted and attached to the lower end. The automatic pipette mechanism 5b is connected to a syringe pump (not shown), and performs suction and discharge according to the operation of the syringe pump controlled by the control unit. The automatic pipette mechanism 5b attaches a disposable pipette tip set in advance to the sample rack 10 to the lower end, moves to a predetermined position to suck or discharge liquid, and moves to the position of the passage 18 after a predetermined operation. To remove the disposable pipette tip. By such an operation, the automatic pipette mechanism 5b performs functions such as sucking target cells from the test tube after separating the specific gravity of blood, and discharging the sucked cells to another test tube for washing. Take on.

The operating arm 5c is a suction mechanism, and a disposable pipette tip is fitted and attached to the lower end. A waste liquid tube (not shown) connected to a waste liquid bottle 15 installed at the lower part of the housing 2 is connected to the suction mechanism 5c. Further, the operation arm 5c is connected to a suction pump (not shown), and performs only the suction operation, and is used when suctioning and discharging unnecessary components and cleaning liquid after the specific gravity separation.
Note that the operation arms 5a, 5b, and 5c do not have to be arranged in the order shown, and may be arranged in any order according to the function, wiring, and the like.

  A waste liquid bottle 15, a cleaning liquid bottle 16, and a trash can 17 are disposed at the bottom of the housing 2. A passage 18 is provided in the upper part of the trash can 17, and disposable pipette tips and test tubes that have been used are put into the trash can 17 through the passage 18. In the housing 2 of the automatic blood separation apparatus 1, a portion that accommodates the trash can 17, the waste liquid bottle 15, and the cleaning liquid bottle 16 is provided with a partition plate and other portions, and a door is provided on the front panel of the housing. It is also preferred that the user of the device can be arbitrarily opened and closed. A damper 19 is provided at the lowermost part of the casing, and the air that has passed through the centrifuge 3 is discharged to the outside through the exhaust port 9 after the wind direction and the wind speed are adjusted by the damper 19.

  A cleaning liquid reservoir 20 is provided between the centrifuge 3 and the sample storage unit 4. The cleaning liquid reservoir 20 is a cylindrical liquid reservoir, and is provided with a supply port 21 in the vicinity of the bottom surface. The supply port 21 is connected to the cleaning liquid bottle 16 by a tube, and the cleaning liquid is discharged from the supply port 21 in accordance with a command from the control unit, and the cleaning liquid is supplied to the cleaning liquid reservoir 20. The automatic pipette mechanism 5b or the suction mechanism 5c sucks the cleaning liquid supplied to the cleaning liquid reservoir 20 as necessary.

  FIG. 2 is a diagram schematically showing the inside of the side surface of the automatic blood separation apparatus 1. The lower part of the casing 2 projects to the front side rather than the upper part, and an openable / closable small door (FIG. 3) for inserting and removing articles to and from the control panel (FIGS. 3 and 30) and the sample storage unit 4 is provided on the upper surface of the projecting part. , 31).

  Housed in the upper part of the housing 2 are a support 13 assembled with a drive unit 6 that supports each arm of the operation unit 5 and an X-axis rail 14. The drive unit 6 does not move in the Z-axis direction. On the other hand, each operation arm (the operation arm 5c in the drawing) includes an electric slider and is movable in the Z-axis direction while being supported by the drive unit 6. For example, a spiral groove that engages with each other is provided on the outer surface of the operation arm 5c and the inner surface of the drive unit 6, and the operation arm 5c rotates to move in the Z-axis direction relative to the drive unit 6. The operation unit 5 includes three operation arms 5a, 5b, and 5c, and each operation arm is independently movable in the Z-axis direction.

  A partition panel 22 is provided inside and on the back side of the upper portion of the housing 2, and a control unit (not shown) including a substrate is housed on the back surface of the partition panel 22. The vent hole 8 is provided so as to extend before and after the partition panel 22, and clean air that has passed through the HEPA filter is supplied to both the front surface and the back surface of the partition panel 22.

  At the lower part of the housing 2, a centrifuge 3, a sample storage unit 4, a damper 19, a trash can 17, and a cleaning liquid bottle 15 are arranged. Inside the sample storage unit 4, the sample rack 10 is placed on a rack rail 11 extending in the X-axis direction, and can move on the rack rail 11 in the X-axis direction. In other words, by combining the movement of the drive unit 6 in the Y-axis direction and the movement of the sample rack 10 in the X-axis direction, the operation unit 5 can move a sample or test tube placed at an arbitrary position of the sample rack 10 Access to pipette tips. In addition to the movement of the sample rack 10 in the X-axis direction, or in place of the movement of the sample rack 10 in the X-axis direction, the movement of the operation unit 5 and the drive unit 6 along the X-axis rail 14 is used. You can also.

  FIG. 3 is a schematic view showing the upper surface of the automatic blood separation apparatus 1. A control panel 30 is provided on the front upper surface of the housing 2. Further, an openable / closable small door 31 is provided for putting an article into and out of the sample storage unit 4. Other configurations have been described above. The centrifuge 3 will be described in detail below.

  FIG. 4 is a schematic diagram showing the centrifuge 3 (stopped) of the automatic blood separation apparatus of the present invention. The centrifuge 3 is a swing rotor type centrifuge, and the rotor 41 is supported by a rotating shaft 42 connected to a motor 40. The rotor 41 has buckets 43 that can accommodate test tubes 44 at both ends. In the example of the drawings, only two buckets 43 are shown for easy understanding, but the number of buckets 43 may be four, six, eight, etc. according to a known configuration. Further, the bucket 43 may be configured to accommodate one test tube in each bucket, or may be configured to accommodate two or more. However, for imaging (described later), it is preferable to have a configuration in which only one test tube does not overlap in the rotational radius direction.

  A protective wall surface 45 is provided on the peripheral side surface of the rotor 41. The protective wall surface 45 is made of a material such as stainless steel having a vinyl chloride lining on the inner surface, and is configured to cover the entire periphery of the rotor 41. The protective wall surface 45 has a see-through window 46, and the see-through window 46 is sealed with transparent glass or the like. An imaging device 47 capable of imaging the inside of the centrifuge 3 is disposed outside the fluoroscopic window 46. The imaging device 47 is, for example, a digital video camera. Further, a light source 48 is disposed at a position facing the imaging device 47 across the bucket 43 inside the centrifuge. The light source 48 can supply light in the direction of the imaging device 47. The surface of the side wall of the bucket 43 that faces the imaging device 47 and the light source 48 is at least partly made of a transparent material, or at least partly provided with an opening, and the test tube 44 inserted into the bucket 43. Can be photographed by the imaging device 47.

  A lid surface 49 is provided on the upper surface of the centrifuge 3, and the lid surface 49 is fixed to the protective wall surface 45 with screws or the like. The lid surface 49 is provided with an opening 50. The opening 50 is an opening having a size through which a pipette tip and a test tube can pass. For example, a circular hole having a diameter of about 30 mm to 60 mm or a square hole having a side length of about 30 mm to 60 mm. The opening 50 may be provided anywhere as long as it is a position directly above the bucket 43, that is, a position on the lid surface 49 corresponding to the rotation trajectory of the bucket 43. In the illustrated embodiment, the cover surface 49 is provided in the depth direction (see FIGS. 3 and 50). However, it is also preferable to provide the opening 50 on a line connecting the imaging device 47 and the light source 48. Although the number of the openings 50 in the lid surface 49 may be one or plural, it is preferably one.

  In the automatic blood separation apparatus of the present invention, the test tube, various samples, and the liquid are put into and out of the centrifuge in principle only through the opening provided on the lid surface. By adopting such a configuration, it is possible to prevent dust and liquid from entering the inside of the centrifuge from the inside, and to prevent contamination in the apparatus due to splashing of the splash from the inside of the centrifuge.

  An exhaust port 51 is provided on the bottom surface of the centrifuge 3. The exhaust port 51 communicates with the damper 19 (FIGS. 1 and 2). In addition to air, the contents of the test tube are also discharged from the exhaust port 51 if the test tube is damaged. While the centrifuge 3 is rotating, the opening 50 on the lid surface serves as an intake port, and an air flow from the opening 50 to the exhaust port 51 on the bottom surface is formed. As described above, since the air that has passed through the HEPA filter 7 is supplied from the vent 8 to the automatic blood separator 1, the air that passes through the centrifuge 3 is only the air that has passed through the HEPA filter. That is, the entire interior of the housing 2 including the centrifuge 3 can be filled with only air passing through the HEPA filter, and the environment is equivalent to a clean room of cleanliness class 3 (JIS B 9220, ISO 14644-1). be able to.

  FIG. 5 is a schematic view showing the centrifuge 3 (during rotation) of the automatic blood separation apparatus of the present invention. During rotation, the bucket 43 that accommodates the test tube 44 is swung outward by centrifugal force and rotates in a state of reaching the horizontal. The number of revolutions and time can be set arbitrarily. For example, when lymphocytes are to be separated from whole blood, use a specific gravity liquid with a specific gravity of 1.077 and 1500 rotations in a 15 mL test tube (diameter 18 mm). , Centrifugation conditions for 35 minutes. During rotation, the imaging device 47 and the light source 48 are not operating.

  FIG. 6 is a schematic diagram showing a state during imaging by the imaging device 47. The rotation is stopped at the time of imaging, and the imaging device 47 images the test tube 44 through the fluoroscopic window 46. Since the test tube 44 is illuminated from behind by the light source 48, a bright and clear image can be obtained.

  FIG. 7 is a control block diagram of the automatic blood separation apparatus of the present invention. The control unit 100 includes a drive unit motor driver 101, a rack rail motor driver 102, a test tube holding mechanism motor driver 103, an automatic pipette mechanism motor driver 104, a suction mechanism motor driver 105, an X-axis rail motor driver 106, and a syringe pump motor driver 107. And controls the position of the drive unit 6, the sample rack 10, the test tube holding mechanism 5a, the automatic pipette mechanism 5b, the suction mechanism 5c and the operation of the syringe pump. The imaging device driver 108 controls the operation of the imaging device 47. The centrifuge stepping motor driver 109 controls the speed of the motor 40 of the centrifuge 3 and the position of the bucket 43.

  Among the above, the drive unit motor driver 101, the rack rail motor driver 102, the test tube holding mechanism motor driver 103, the automatic pipette mechanism motor driver 104, the suction mechanism motor driver 105, and the X-axis rail motor driver 106 are positioning mechanisms. That is, the position of the drive unit 6 (operation arm 5) in the X-axis and Y-axis directions, the position of the sample rack 10 in the X-axis direction, and the Z-axis direction of the test tube holding mechanism 5a, automatic pipette mechanism 5b, and suction mechanism 5c. Are controlled by these motor drivers.

  In addition, the automatic pipette mechanism motor driver 104 and the syringe pump driver 107 cooperate to suck or discharge the liquid corresponding to the operation amount of the syringe pump with a disposable pipette tip attached to the tip of the automatic pipette mechanism 5b.

  The suction pump 120, the cleaning liquid pump 121, and the fan 123 are also controlled by the control unit 100. The suction pump 120 is connected to the suction mechanism 5c and performs a suction operation. The cleaning water pump 121 supplies the cleaning liquid to the cleaning liquid reservoir 20 according to a command from the control unit.

  Various parameters for specific gravity separation, such as centrifugation conditions (speed, time), the amount of specific gravity liquid, and the like are input from the control panel 30 and input to the main microcomputer. In response to the input information, a command is sent from the main microcomputer to each motor driver through each motor control microcomputer. In addition, the suction pump, the cleaning liquid pump, and the fan are activated. The stepper motor of the centrifuge is operated through the motor driver 109.

  Each electric slider and stepping motor is provided with a position confirmation mechanism, and the current position of each part in operation is transmitted to the main microcomputer.

  Next, an outline of a series of operations for separating cells from blood using the automatic blood separation device of the present invention will be described with reference to FIGS. In the automatic blood separation apparatus of the present invention, blood separation operations can be performed roughly by two methods. One is a method of executing the step of overlaying blood and specific gravity liquid inside the centrifuge, and the other is a method of executing the step of overlaying blood and specific gravity solution on the sample rack.

  FIG. 8 is an operation flowchart in the case of executing the step of layering blood and specific gravity liquid inside the centrifuge. First, before the step S1, when the operator turns on the power, the control unit main microcomputer, the fan 123, the suction pump 120, and the cleaning liquid pump 121 are started up. The operator inputs various information from the control panel 30. A predetermined initialization process is performed in step S1 of FIG. Initialization processing includes confirmation of various parameters based on information input from the control panel 30, various liquids such as samples placed on the sample rack 10, specific gravity liquid, and various equipment (U bottom test tube, V bottom test tube) The position and quantity of the disposable pipette tips, etc.), the movement of the operating unit 5 and the bucket 43 of the centrifuge 3 to a predetermined initial position, and the like. Also, a predetermined amount of specific gravity solution is dispensed into the required number of U-bottom test tubes, and a predetermined amount of washing solution (water or PBS or the like) is also dispensed into the V-bottom test tubes.

  In step S2, a process of moving the U-bottom test tube and the V-bottom test tube from the sample rack 10 to the centrifuge 3 is performed. In this step, a U-bottom test tube into which a specific gravity liquid was dispensed in the previous step and a V-bottom test tube for collecting a target fraction after specific gravity separation were inserted into the bucket 43 of the centrifuge 3 and set. It is a process to do.

  As a specific operation in step S2, the operation arm 5a (test tube holding mechanism) is moved to the sample rack by the cooperative operation of the sample rack 10 (rack rail electric slider 11) and the drive unit 6 (drive unit electric slider 6). Move to a position on the target test tube placed on the. Subsequently, the test tube holding mechanism electric slider 5 a is operated, and the operation arm 5 a is lowered to a position where the lower end of the operation arm 5 a is in contact with the mouth of the test tube placed on the sample rack 10. After fitting and holding the test tube at the lower end of the operation arm 5a, the test tube holding mechanism electric slider 5a is actuated again, and the operation arm 5a is raised. Subsequently, the drive unit electric slider 6 is operated, and the operation arm 5 a moves to the position just above the opening 50 of the centrifuge 3. At the same time, among the tubes of the bucket 43 of the centrifuge 3, the centrifuge stepping motor is arranged so that the tube to receive the test tube conveyed by the operation arm 5 a is positioned directly below the opening 50 of the centrifuge 3. 40 operates. Next, the test tube holding mechanism electric slider 5a is operated to move down to the position of the bucket 43 while holding the test tube, and the holding of the test tube by the bucket 43 is released. Thereafter, the test tube holding mechanism electric slider 5a is operated to raise the operating arm 5a, and then the driving unit electric slider 6 is operated to return to the original position. By repeating this operation, predetermined U-bottom test tubes and V-bottom test tubes are moved into the centrifuge 3.

  In step S3, a blood overlaying process is performed. This step is a step of overlaying blood (sample) on the specific gravity liquid-containing U-bottom test tube set in the centrifuge in the previous step S2. In addition, when performing blood separation work by human hands, the specific gravity separation is performed after diluting whole blood about twice, but according to the apparatus of the present invention, a dilution step is unnecessary, and it is possible to remove from the patient or subject. The collected blood can be subjected to separation work as it is.

  As a specific operation of step S3, first, the operating arm 5b (automatic pipette mechanism) is sampled by the cooperative operation of the sample rack 10 (rack rail electric slider 11) and the drive unit 6 (drive unit electric slider 6). It moves over the unused disposable pipette tip placed on the rack, and then the automatic pipette mechanism electric slider 5b is actuated and lowered, and one of the disposable pipette tips placed on the sample rack 10 is moved. Attach to the tip and rise again. Subsequently, the cooperative movement of the sample rack 10 (rack rail electric slider 11) and the drive unit 6 (drive unit electric slider 6) causes the operation arm 5b (automatic pipette mechanism) to be placed on the sample rack. Move to a predetermined location on the test tube to be accommodated. Next, the automatic pipette mechanism electric slider 5b is actuated to descend into a test tube containing blood. Next, a predetermined amount of blood is sucked into the pipette tip by the operation of the syringe pump electric slider. The operating arm 5b is lifted with the blood held in the pipette tip, and then the driving unit electric slider 6 and / or the X-axis rail electric slider 14 is operated, and the operating arm 5b is moved to the true position of the opening 50 of the centrifuge 3. Move to the top. At the same time, the centrifuge stepping motor 40 operates so that one of the U-bottom test tubes accommodated in the bucket 43 in the centrifuge 3 is positioned directly below the opening 50 of the centrifuge 3. Next, the automatic pipette mechanism electric slider 5b operates to lower the operation arm 5b to a predetermined position. Next, the blood held on the pipette tip is discharged by the operation of the syringe pump electric slider, thereby superposing the blood on the specific gravity liquid previously placed in the U-bottom test tube. Thereafter, the automatic pipette mechanism electric slider 5b is operated to raise the operating arm 5b, and then the driving unit electric slider 6 and / or the X-axis rail electric slider 14 are operated to return to the original positions. Before returning to the original position, the pipette tip may be detached on the passage 18 by stopping on the passage 18. This operation is repeated so that a predetermined number of blood layers are stacked in different U-bottom test tubes.

  Step S4 is a specific gravity separation step. As a result of step S3, the bucket 43 of the centrifuge 3 holds a U-bottom test tube in which a specific gravity liquid and blood are layered. After performing balance adjustment as necessary, centrifugation is performed at a rotation speed (acceleration) and time according to a predetermined protocol. Specifically, the centrifuge stepping motor 40 operates according to a command from the control unit 100.

  In step S5, the target layer is recognized and collected. In step S5, the U-bottom test tube containing the blood separated in specific gravity is imaged by the imaging device, the interface position of the sample separated in specific gravity in the test tube is detected by image analysis, and the detected interface position information is obtained. In this step, the target layer is collected by suction and transferred to a V-bottom test tube. The separation method using the apparatus of the present invention is characterized by this step S5, and only the layer containing the target cell is collected more accurately and reliably by photographing each test tube after the specific gravity separation. Can do.

  As a specific operation in step S5, after the specific gravity separation process in step S4 is completed, the illumination 48 is turned on, and the bucket 43 that accommodates the test tube to be imaged is operated by the operation of the stepping motor 40. It is moved substantially in front (between the illumination 48 and the imaging device 47). Next, the imaging device 47 is operated to photograph the test tube. The photographed image is sent from the imaging device driver 108 to the main microcomputer, and the interface position of the liquid in the test tube (position of the target layer) is calculated by image analysis. Next, the automatic pipette mechanism 5b equipped with an unused pipette tip is operated, and the target layer is sucked and collected by the automatic pipette mechanism 5b. The suction amount of the automatic pipette mechanism 5b at this time is determined by converting the interface position information calculated above into the operation amount of the syringe pump. The collected target layer is discharged to a V-bottom test tube containing a cleaning liquid previously stored in the bucket 43. The automatic pipette mechanism 5b does not move in the XY plane from suction to discharge, and the transfer operation from the U-bottom test tube to the V-bottom test tube proceeds as the bucket 43 moves. This operation is repeated for each blood sample, and the target layer of each blood sample separated by specific gravity is transferred to each separate V-type test tube.

  Step S6 is a step of mixing the collected cells and the washing solution in a V-shaped test tube. Step S6 may be performed simultaneously with step S5. Specifically, the operation in step S6 is performed by positioning the automatic pipette mechanism 5b in the V-shaped test tube, operating the syringe pump, and repeating suction and discharge several times.

  Step S7 is a light centrifugation process. The collected cells and the washing solution mixed in step S6 are lightly centrifuged in step S7 to precipitate the collected cells, and the washing solution and the residue mixed with the collected cells are used as a supernatant. If necessary, adjust the balance before centrifugation. Specifically, in the operation of step S7, the centrifuge stepping motor 40 operates according to a command from the control unit 100. As a result of step S7, the target collected cells are precipitated at the bottom of the V-bottom test tube, and the washing solution and the residue become the supernatant.

  Step S8 is a supernatant suction / discard process. In this step, the layer separated as the supernatant in step S7 is discarded. Specifically, the operation in step S8 is as follows. First, one of the V-bottom test tubes containing the washing liquid layer (supernatant) and the collected cell layer (precipitation) is moved directly below the opening 50 of the centrifuge 3 by the operation of the centrifuge stepping motor 40. At the same time, the operation arm 5 c (suction mechanism) is moved onto the opening 50 by the operation of the drive unit 6 and / or the X-axis rail electric slider 14. A new pipette tip is attached to the tip of the operating arm 5c. Next, the suction mechanism electric slider 5c is actuated to lower the operation arm 5c, and the supernatant in the V-bottom test tube is sucked. The suction mechanism of the operating arm 5c is connected to the waste liquid bottle 15 by a tube, and the sucked supernatant is discarded to the waste liquid bottle 15 through the tube. After sucking a predetermined amount, the suction mechanism electric slider 5c is actuated again to raise the operating arm 5c. If necessary, the pipette tip at the tip of the operating arm 5c is replaced. The exchange of pipette tips can be performed in the same procedure as in the case of the automatic pipette mechanism 5b in step S3. Repeat this action and aspirate the supernatant from all V-bottom test tubes. As a result of step S8, collected cells (which may be suspended in a small amount of washing solution) remain in the V-bottom test tube.

  Step S9 is a step of taking the U-bottom test tube and the V-bottom test tube from the centrifuge into the sample rack. In this process, in addition to the V-bottom test tube containing the target collection cell obtained in step S8, all the test tubes existing in the centrifuge 3 are taken out from the centrifuge. In step S9, the U-bottom test tube and the V-bottom test tube are moved to the sample rack 10 by using the operation arm 5a (test tube holding mechanism) by executing the operation of step S1 in reverse order.

  Step S10 is collection of the target cells, and this step is performed manually. As a result of step S9, the sample rack 10 is placed with the V-bottom test tube in which the target cells are collected. Target cells are obtained from the V-bottom test tube.

  The above is an outline of the operation, and various operations not described are performed as necessary. For example, pipette tip replacement, cleaning, disposal, suction, bucket position adjustment, and the like are executed at each stage.

  FIG. 9 is an operation flowchart in the case of executing the step of overlaying blood and specific gravity liquid on the sample rack. Except for step S20 and step S30, the operation is the same as that described with reference to FIG.

  Step S <b> 20 is a blood layer process performed on the sample rack 10. As a specific operation of step S20, first, the drive unit electric slider 6 and / or the X-axis rail electric slider 14 is operated, and the operation arm 5b (automatic pipette mechanism) is placed on the unused disposable pipette tip of the sample rack 10. Move. Subsequently, the automatic pipette mechanism electric slider 5b is operated to lower the operation arm 5b, and a pipette tip is attached to the tip. Next, the operating arm 5b moves to the position of the test tube (collecting blood vessel) that stores the blood sample previously placed on the sample rack 10, and the syringe pump operates to enter the pipette tip at the tip of the automatic pipetting mechanism 5b. A predetermined amount of blood sample is aspirated. With the blood held in the pipette tip, the driving unit electric slider 6 and / or the X-axis rail electric slider 14 is operated again, and the U bottom test with specific gravity liquid placed at another position on the sample rack 10 is operated. The operating arm 5b is moved to the position of the tube. After the operation arm 5b moves onto the U-bottom test tube, the syringe pump operates to discharge blood into the U-bottom test tube, thereby superimposing blood on the specific gravity liquid. After the blood is discharged, the operating arm 5b moves onto the passage 18 and removes the pipette tip. By repeating this operation, each blood sample is layered on the specific gravity liquid in each U-bottom test tube on the sample rack 10.

  Step S <b> 30 is a step of moving the U-bottom test tube and the V-bottom test tube in which the specific gravity solution and the blood are layered, into the centrifuge 3. It is also preferable to dispense a washing solution into the V-bottom test tube in advance. The specific operation of moving the U-bottom test tube and the V-bottom test tube can be performed in the same manner as described in step S2 of FIG.

  Each operation described above is performed based on a command from the main microcomputer of the control unit to each motor control microcomputer. Further, the position information of each part is fed back from each part to the main microcomputer, and the operation is executed while checking the current position of each part. Also,

  According to the above-described series of operations, it is possible to automatically perform the blood separation operation without using a human hand, and there is a great advantage in terms of operation accuracy and operation load. In addition, since most of the work is performed inside the centrifuge, the risk of contamination and contamination is low, and a safe method can be provided.

DESCRIPTION OF SYMBOLS 1 Automatic blood separator 2 Case 3 Centrifuge 4 Sample storage part 5 Operation part 6 Drive part 8 Vent 10 Sample rack 11 Rack rail 12 Y axis rail 14 X axis rail 19 Damper 40 Motor 41 Rotor 43 Bucket 44 Test tube 45 Protective wall 46 Transparent window 47 Imaging device 48 Light source 49 Cover surface 50 Opening

Claims (6)

A housing and a control unit provided inside or outside the housing;
A centrifuge, a sample storage unit, an operation unit, and a drive unit are provided inside the housing.
The operating unit has at least a test tube holding mechanism and an automatic pipette mechanism,
The drive unit moves the operation unit in an upper space of the centrifuge and the sample storage unit,
The centrifuge has a protective wall surface provided with a see-through window on a part of a peripheral side surface, and an imaging device capable of taking an image of the inside of the centrifuge through the see-through window is provided outside the see-through window. And
The automatic blood separation device, wherein the control unit controls the centrifuge, the operation unit, the driving unit, and the imaging device. The automatic blood separation device according to claim 1, wherein the centrifuge is a swing rotor type centrifuge. The automatic blood separation device according to claim 1 or 2, wherein a light source is provided inside the centrifuge, and the imaging device and the light source are opposed to each other with a bucket of the centrifuge interposed therebetween. The housing has a vent, the vent has a HEPA filter, and the centrifuge has a lid surface fixed to the upper surface and a bottom surface, and each of the lid surface and the bottom surface has an opening. The automatic blood separation device according to any one of claims 1 to 3, wherein only air that has passed through a HEPA filter is ventilated through the opening. The automatic blood separator according to claim 4, wherein the articles are taken into and out of the centrifuge only through an opening on a lid surface of the centrifuge. A test tube holding mechanism capable of holding a test tube; an automatic pipette mechanism that attaches a pipette tip to a tip and performs suction and discharge; a suction mechanism that has a tube connected to a waste bottle and performs suction only; The automatic blood separation device according to any one of claims 1 to 5, comprising:

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