Patent Grant
12220712
This application is a 371 application of the International PCT application serial no. PCT/JP2020/017761, filed on Apr. 24, 2020, which claims the priority benefits of Japan Patent Application No. 2019-119351, filed on Jun. 27, 2019. The entirety of the above-mentioned patent application is hereby incorporated by reference herein and made a part of this specification.
The present invention relates to a centrifuge that automatically washes living cells such as blood cells or the like by using centrifugal force, and more particularly to a centrifuge that can precisely adjust the remaining amount of a supernatant discharged from a plurality of test tubes in a supernatant discharging step (remaining amount of decanting).
In a supernatant discharging step of a conventional cell washing centrifuge, a test tube holder is sucked by a magnetic device, a test tube is rotated while being held in a substantially perpendicular direction, and the supernatant in the test tube is discharged by a centrifugal force. A technique of Patent literature 1 is known as this cell washing centrifuge that discharges a supernatant. In Patent literature 1, the cell washing centrifuge includes: a plurality of test tube holders that are rotatably mounted on a rotor in a circular row and rotated in an outer horizontal direction of the circular row by a centrifugal force generated by the rotation of the rotor; a cleaning liquid distribution element that supplies a cleaning liquid into a plurality of test tubes that are mounted on the inner side of the rotor; and a magnetic element (holding part) that sucks the test tube holder vertically or at a nearly vertical angle by a magnetic attraction force generated by energization of a magnetic coil. The cleaning liquid distribution element has a nozzle (cleaning liquid injection port) installed radially from the outer periphery of a bottom surface of a container whose inner surface has a conical shape, and the cleaning liquid distribution element uniformly divides the cleaning liquid injected by the centrifugal force from the center of the cleaning liquid distribution element that rotates with the rotor, and supplies the cleaning liquid into the plurality of test tubes held by the test tube holders through the nozzle. A cleaning process of the cell washing centrifuge, including a cleaning liquid injection step, a centrifuging step, a supernatant discharging step, and an agitating step, is automatically performed in sequence. Of these, in the supernatant discharging step, the test tube holder is held on the rotor by the magnetic element in a state of being tilted outward at a small angle from the vertical direction, and the rotor is rotated at a low and constant speed. Thereby, the supernatant of the cleaning liquid is discharged from an upper opening of the test tube by the centrifugal force.
In the supernatant discharging step of the conventional cell washing centrifuge, the test tube holder is sucked to hold the test tube in a substantially vertical state, and the supernatant in the test tube is discharged by the centrifugal force when the rotor is accelerated and settled. Thus, the discharge amount of the supernatant is determined by a rotation speed when the rotor is settled and a centrifugal time including an acceleration time. In this way, the conventional supernatant discharge control depends greatly on the rotation speed control of the motor, and thus a highly accurate motor control technique such as a technique that does not overshoot the rotation speed at the time of settling or the like is required. In addition, after the supernatant discharging step is completed, it is difficult to remain the cleaning liquid in the test tube in an amount desired by a user, that is, it is difficult to finely control the discharge amount of the supernatant.
The present invention has been made in view of the above background, and an object of the present invention is to provide a centrifuge that can precisely control the discharge amount of a supernatant. Another object of the present invention is to provide a centrifuge that performs a first decanting operation in which a supernatant discharging step is performed in a state that a test tube holder is sucked, and a second decanting operation in which the test tube holder is made to swing by releasing the suction state of the test tube holder using a holding part during rotation of a rotor. Still another object of the present invention is to provide a centrifuge that can adjust the amount of a cleaning liquid remaining in a test tube by moving a timing for releasing the suction of a test tube holder during the supernatant discharging step (during the rotation of the rotor).
Typical features of the invention disclosed in the present application are described as follows. According to one feature of the present invention, a cell washing centrifuge includes: a motor; a rotor that is mounted on a drive shaft of the motor; a plurality of test tube holders that are arranged side by side in a circumferential direction of the rotor and are rotatable (can agitate) in a radial direction by a centrifugal force generated by the rotation of the rotor; a cleaning liquid distribution element that is held in the rotor and supplies a cleaning liquid into a plurality of test tubes held by the test tube holders; a holding part capable of preventing the rotation of the test tube holder; and a control device for controlling rotation of the motor and the operation of the holding part. In the cell washing centrifuge, the control device performs the following steps: a cleaning liquid injection step of injecting the cleaning liquid into the test tube by the cleaning liquid distribution element during the rotation of the rotor; a centrifuging step of rotating the test tube holders by the centrifugal force generated by the rotation of the rotor; and a supernatant discharging step of rotating the rotor in a state in which the test tube holders are held by the holding part and discharging the supernatant of the cleaning liquid from the test tube. In the supernatant discharging step, during the rotation of the rotor, particularly, during the acceleration of the rotor, by releasing the holding state of the test tube holders held by the holding part, the test tube holders can be made to swing from the fixed state and the discharge of the supernatant can be stopped halfway. In this way, in the present invention, the amount of the supernatant remaining in the test tube can be adjusted according to a timing for releasing the test tube holders from the holding state.
According to another feature of the present invention, the supernatant discharging step includes control of “acceleration, settling, and deceleration” of the rotor. Furthermore, when the holding of the test tube holders by the holding part is released during the acceleration of the rotor, the rotation of the rotor is controlled to be decelerated without being settled thereafter. When the holding of the test tube holders are released during the acceleration of the rotor, the amount of the cleaning liquid remaining in the test tube can be adjusted according to the rotation speed of the rotor when the holding of the test tube holders are released. With this configuration, the residual amount of the cleaning liquid can be adjusted to an amount desired by the user by changing the timing for releasing the holding of the test tube holders back and forth. The holding part includes an electromagnet, and the control device fixes (prevent swinging of) the test tube holders by sucking the test tube holders which includes a magnetic material by the electromagnet. With this configuration, the suction or the release of the test tube holders can be easily controlled according to an electric signal from the control device. Furthermore, a stopper that restricts the agitating angle of the test tube holders with respect to the drive shaft during the centrifugation is formed on the rotor, and a maximum agitating angle during the centrifugal separation operation is constant.
According to still another feature of the present invention, a centrifuge includes: a rotor rotated by a motor; a cleaning liquid distribution element that injects a cleaning liquid into a test tube mounted on the rotor during the rotation of the rotor; an agitating angle changing part that can switch the agitating angle of the test tube with respect to the rotor; and a control device for controlling the rotation of the motor, the injection of the cleaning liquid, and the change of the agitating angle. The control device performs two types of decanting operations. In a first decanting operation, the rotor is rotated in the order of “acceleration, settling, and deceleration” in a state that the agitating angle of the test tube is restricted, and the supernatant of the cleaning liquid is discharged from the test tube. In a second decanting operation, the rotor is accelerated, the restriction on the agitating angle is released during the acceleration, and then the rotor is decelerated. That is, the second decanting operation does not include the “settling” operation of the rotor. The amount of the cleaning liquid remaining in the test tube after the second decanting operation can be easily adjusted according to the switching timing of the agitating angle during the acceleration of the rotor. The second decanting operation is performed after the first decanting operation, and is preferably performed as the final decanting operation. Furthermore, a switching timing of the agitating angle during the second decanting operation can be set in advance by the user, and thus the user can freely set the amount of the cleaning liquid remaining in the test tube.
According to the present invention, the amount of the supernatant discharged from the test tube (decant amount) can be controlled by adjusting the timing for releasing the suction of the test tube holders in the supernatant discharging step, that is, the rotation speed at the time of releasing the suction. In particular, different from the conventional adjustment of the decant amount which is performed by agitating (swinging) the test tube holders during the acceleration of the rotation of the rotor and depends on the rotation speed at the time of the settling and the time, the decant amount can be precisely adjusted by the control device 10. Furthermore, in the conventional control, the remaining amount of the supernatant after decanting is precisely remained only in a small amount (less than 1 mL), but in this method, by freely changing the timing for releasing the suction, the remaining amount of the supernatant after decanting can be precisely remained even in a large amount (1 mL or more).
(A) of
Hereinafter, an embodiment of the present invention is described in detail with reference to the drawings. Note that, in all the diagrams for describing the embodiment, the members having the same function are designated by the same reference signs, and the repeated description thereof is omitted.
The rotor 20 is a dedicated rotor for washing cells, and has a plurality of (for example, 24) test tube holders 31 arranged side by side at equal intervals in a circumferential direction when viewed from the upper surface. The test tube holder 31 is held in a centrifugal direction (radial direction) in a swingable (rotatable) manner by pivoting an inner peripheral side surface by a rotor plate 22 (reference sign is shown in
The rotor 20 includes a holding part 27 for holding a longitudinal central axis of the test tube holder 31 vertically or at a nearly vertical agitating angle. The holding part 27 keeps the metal test tube holders 31 in a non-swingable state by sucking the metal test tube holder 31 by a magnetic force, and uses a magnetic element such as an electromagnet or the like. The holding part 27 can electrically switch between a suction state (fixed state or non-swingable state) and a released state (swingable state) of the test tube holder 31. The holding part 27 functions as a so-called angle rotor having a negative swinging angle when the test tube holder 31 is in the suction state, and functions as a so-called swing rotor when the test tube holder 31 is in the released state. A swinging angle θ of the test tube in the released state is about 45 degrees, which is described later in
The rotor 20 for cell washing is detachable with respect to the drive shaft 9. Therefore, the drive shaft 9 can also be equipped with a normal angle rotor or a normal swing rotor that cannot supply a cleaning liquid during the rotation. When the rotor 20 for cell washing as in the example is mounted on the drive shaft 9, a cleaning liquid distribution element 25 is mounted on an upper portion of the rotor 20, and a cleaning liquid supply pipe 18 arranged in the door 6 is used to supply a liquid such as cleaning liquid or the like into the test tube 40 described later in
A nozzle 19 serving as an outlet of the cleaning liquid supply pipe 18 is arranged on a rotation axis A1 at an upper portion of the cleaning liquid distribution element 25, and the liquid falling from the nozzle 19 flows into a cleaning liquid inflow port 25a located on the upper side of the cleaning liquid distribution element 25. The cleaning liquid inflow port 25a is on the rotation axis A1 at the upper portion and forms a space that is connected to a cleaning liquid passage 25b having a conical internal space. An outer edge portion of the cleaning liquid passage 25b is divided in the circumferential direction, and a plurality of cleaning liquid injection ports 25c extending in the radial direction (see
A pump (not shown) is coupled to an outer end portion (an end portion apart from the nozzle 19) of the cleaning liquid supply pipe 18 that supplies the cleaning liquid to the cleaning liquid distribution element 25. By turning on (ON) an operating power of the pump by the control device 10, a cleaning liquid 17 can be supplied from an external cleaning liquid tank (not shown) to the nozzle 19 located at the upper portion of the centrifuge 1 through the cleaning liquid supply pipe 18. In a cleaning liquid injection step described later, the cleaning liquid ejected downward from the nozzle 19 enters the central portion of the cleaning liquid distribution element 25 that rotates integrally with the rotor 20 at a high speed, is distributed to flow to the outer periphery by the centrifugal force in the cleaning liquid distribution element 25, is branched into each of flow paths having the same number (24) as that of the test tubes 40 held in the tube holders 31, and is vigorously injected into each test tube 40 from the cleaning liquid injection ports 25c of the cleaning liquid distribution element 25.
A bowl-shaped bottom surface portion 23 is formed at the lower portion of the rotor 20. The bottom surface portion 23 is a container for receiving the cleaning liquid spilled without entering the test tube 40 and also serves as a stopper for restricting the swinging angle of the test tube holder 31. That is, the test tube holder 31 that holds the test tube rotates in a radial horizontal direction of the circumference of the rotor 20 and tilts until the lower portion (a holding bottom portion 31c described later) of the test tube holder 31 contacts an outer edge portion of the bottom surface portion 23. In the contact state, the sample such as the blood cells or the like in the test tube 40 is centrifuged.
Because the cleaning liquid is injected in a state in which the rotor 20 is rotated and the excess cleaning liquid is discharged from the inside of the test tube 40, the spilled cleaning liquid is accumulated on a bottom surface portion of the chamber 3. Thus, a drain hose 7 is connected to a portion of the bottom surface of the chamber 3, and a discharge port 7a of the drain hose 7 is arranged extending to the outside of the housing 2. The user collects or discards the excess cleaning liquid (waste liquid) using a hose or the like at the front end of the discharge port 7a.
The test tube holder 31 is a member that holds the test tube 40 made of glass or synthetic resin not to fall when the test tube 40 is stopped or when the centrifugal separation operation is performed. The test tube holder 31 is made of a magnetic material, for example, a stainless alloy that is sucked by a magnet made of SUS430 material. Holding insertion portions 31a and 31b are formed in the middle of a longitudinal direction of the test tube holder 31, and the holding bottom portion 31c that supports a bottom of the test tube 40 is formed at a lower end portion in the longitudinal direction. The holding insertion portions 31a and 31b are portions formed by bending a portion of a metal plate into a ring shape, and the holding bottom portion 31c is a portion that holds the bottom of the test tube 40 by bending a portion of the metal plate cut out by press working radially outward. Each test tube holder 31 is held on an outer peripheral edge of a circular shape holding portion (the rotor plate 22) in a state in which the test tube holder 31 can be swinged by a rotating shaft 30. A torsion spring 32 is arranged on the rotating shaft 30, and when an external force caused by the centrifugal force is not applied to the test tube holder 31, the test tube holder 31 is urged to move to a position shown in
The holding part 27 includes a magnetic element (electromagnet) that generates magnetism by electric power. The holding part 27 includes a disk-shaped upper magnetic member 27a and a lower magnetic member 27b, and is further constituted of a ring-shaped coil (magnetic coil) 27c of an insulated wire installed so as to be clamped between the upper magnetic member 27a and the lower magnetic member 27b. The holding part 27 is fixed to the rotor 20, thus rotating together with the rotor 20. In addition, when the rotor 20 is removed from the drive shaft 9, the holding part 27 is also removed. Wiring of the holding part 27 to the magnetic coil 27c is performed from the bottom surface side of the chamber 3 by a slip ring 16, and an electric current can be supplied to the magnetic coil 27c not only when the rotor 20 is stopped but also when the rotor 20 is rotating. The on or off of the electric current supply is controlled by the control device 10 that has a microcomputer. When the electric current is applied to the magnetic coil 27c, a strong magnetic force can be generated which passes through the upper magnetic member 27a and the lower magnetic member 27b. Because the test tube holder 31 is made of a magnetic material, the test tube holder 31 forms a magnetic circuit together with the upper magnetic member 27a and the lower magnetic member 27b. That is, by supplying the electric current to the magnetic coil 27c, the holding part 27 (the magnetic members 27a and 27b) acts as one magnet and sucks the test tube holder 31 made of the magnetic material.
An outer diameter of the upper magnetic member 27a is larger than that of the lower magnetic member 27b. Accordingly, the suction surfaces of the upper magnetic members 27a and lower magnetic member 27b can hold the test tube holder 31 in a state in which a bottom side of the test tube 40 is slightly tilted inward, in other words, an upper opening is slightly tilted radially outward (agitating angle θ=about −7 degrees) with respect to a vertical line (completely parallel to the rotation axis A1 of the rotor). A labyrinth portion 29 is formed on a bottom surface of the lower magnetic member 27b to limit flow of air between a bearing 15 and the rotor chamber 4.
When the cleaning liquid injection step is performed using this swingable rotor 20, the test tube holder 31 rotates in the outer horizontal direction of the circular row by the centrifugal force generated by the rotation of the rotor 20. In the rotating state as shown in
(A) of
Next, an execution procedure of a cleaning cycle is described with reference to
When the injection of the cleaning liquid is completed in the middle of the acceleration section and the rotation speed of the rotor 20 reaches the set rotation speed R3 of the centrifugal separation operation at time t1, the operation is performed for the set time (centrifugal separation operation time=t2−t1). Here, as shown in the column of {circle around (2)} in
When the rotation of the rotor 20 is stopped at time t3 in
An agitating step is performed after the rotor 20 is stopped at time t4. The agitating step is a step of stirring the remaining cleaning liquid and the sample by agitating the test tube holder a plurality of times in a short time (AGITATE). Here, the rotation speed of the rotor 20 is accelerated to R1, settled for a short time, and then decelerated immediately. The operation of repeating rotation and stop is performed a plurality of times (here, 5 times) in steps of acceleration, settling, and stop. As described above, the cleaning cycle from {circle around (1)} to {circle around (4)} is repeated a plurality of times, for example, about 3 to 4 times, and as shown in
The supernatant discharging step shown in {circle around (3)}-2 of the third cycle has the following four features. (1) During the operation of the rotor 20, the settling section is eliminated, and the operation pattern is set to “acceleration and deceleration” only. (2) The acceleration is started when the test tube holder 31 is sucked by the holding part 27, and the suction of the test tube holder 31 by the holding part 27 (see
As a result of the above control, in the final supernatant discharging step (second decanting operation indicated by {circle around (3)}-2), the discharge of the supernatant from the test tube 40 is interrupted during the acceleration (the timing indicated by the arrow 51). In the embodiment, after the supernatant discharging step ({circle around (3)}-2), the amount of the cleaning liquid remaining in the test tube 40 can be precisely adjusted to a desired amount by adjusting the timing for releasing the test tube holder 31 (rotation speed of the arrow 51).
If it is desired to increase the amount of the cleaning liquid remaining inside the test tube 40, the suction of the test tube holder 31 may be released at a timing earlier than the release timing 51, for example, at a timing 51a. If it is desired to reduce the amount of the cleaning liquid, the suction of the test tube holder 31 may be released at a timing later than the release timing 51, for example, at a timing 51b. The releasing of the suction of the test tube holder 31 can be achieved only by releasing the power supply to the ring-shaped coil 27c, and thus can be easily controlled by the control device 10. In this way, because the release timing 51 is assigned during the acceleration of the rotor 20, the amount of the residual cleaning liquid can be increased by shifting the release timing 51 toward the direction of an arrow 52a (advancing the release timing), and conversely, the amount of residual cleaning liquid can be reduced by shifting the release timing 51 toward the direction of an arrow 52b (delaying the release timing). The adjustment of the amount of the residual cleaning liquid according to the embodiment can also be freely specified by the user. For example, in a case that a standard release timing is 51, if the actual release timing is set to two stages (adjustment levels of the remaining amount +1 and +2) in the direction of the arrow 52a, and similarly, if the actual release timing is set to two stages (adjustment levels of the remaining amount −1 and −2) in the direction of the arrow 52b, the amount of the residual cleaning liquid can be set to a total of five stages. The setting level including these five stages may be configured to be settable by the user from the operation display panel 12. Note that, the number of stages that the release timing can be set is optional, and the release timing may be set continuously variably instead of being set in stages.
When the injection of the cleaning liquid is completed and the rotation speed of the rotor 20 reaches the specified centrifugal rotation speed, the centrifuging step of {circle around (2)} is performed. In the centrifuging step, the operation is performed at a constant speed only for a time set according to the centrifugal rotation speed R3. Here, for example, the rotation speed of the rotor 20 is set to 3000 rpm and the centrifugation is performed for 45 seconds. Accordingly, the blood cells precipitate at the bottom of the test tube 40, and unnecessary substances such as serum or the like remains in a supernatant state (step 62). Next, the control device 10 determines whether the performed centrifuging step is the final cycle of the plurality of cycles (step 63). Here, when the performed centrifuging step is not the final cycle, that is, when the centrifuging step is performed at time t3 or time t8 in
In step 63, because the cleaning operation performed at time t13 is the final cycle among the three cycles, the “supernatant discharging step 2” according to the embodiment is performed in step 66. Here, a magnetic field is generated when the energization of the magnetic coil 27c is turned on (ON), and the test tube holder 31 is sucked and fixed in a substantially vertical state. In the state that the test tube holder 31 is held substantially vertically in this way, the rotor 20 is accelerated, and at an intermediate stage in which the rotation speed of the rotor 20 reaches the specified rotation speed of 400 rpm, that is, at the timing 51 of
When the energization of the magnetic coil 27c is turned off during the acceleration of the rotor 20, the lower portion of the test tube 40 agitates radially outward by the centrifugal force. At this time, because the upper opening surface of the test tube 40 faces the inner peripheral side of the rotor 20, the outflow of the supernatant from the test tube 40 is prevented (step 66). Then, when the lower portion of the test tube 40 remains agitating outward in the radial direction of the rotor 20, and the rotation speed of the rotor 20 is continuously increased and reaches the specified rotation speed of 400 rpm, the control device 10 decelerates the rotor 20.
Next, in the agitating step, by alternately repeating the rotation and stop of the rotor 20 at intervals, or by alternately repeating forward rotation and reverse rotation at intervals, the control device 10 agitates the test tube 40 in the test tube holders 31, and he blood cells that had precipitated and stuck to the bottom of the test tube 40 are dissolved (step 67). Finally, because when the test tube 40 is taken out, water droplets and the like may be attached to an outer wall of the test tube 40, in order to drop the water droplets, the rotor 20 is accelerated to a rotation speed sufficient to drop the water droplets and then stopped. (step 68). By the acceleration and the deceleration in step 68, the blood cells precipitated in the test tube 40 can be positioned at the center of the bottom surface, and as a result, the precipitate is easily taken out from the test tube 40 after the operation is completed. The steps described above complete the cleaning process for performing the blood transfusion tests or the like.
Although the present invention has been described above based on the embodiment, the present invention is not limited to the above-mentioned embodiment, and various modifications can be made without departing from the spirit of the present invention. For example, in the supernatant discharging step of the embodiment described above, the amount of residual cleaning liquid is adjusted by releasing the holding state of the test tube holder during the acceleration and shifting the release timing back and forth. This may be controlled in a manner that the holding state of the test tube holder is released at the time of settling the supernatant discharging step performed in the order of “acceleration, settling, and deceleration”, and the rotation speed at the time of settling is increased or decreased according to the amount of the residual cleaning liquid. In addition, in the embodiment described above, the test tube holder 31 is released during acceleration only in the last cycle among the plurality of cycles, but the test tube holder 31 may also be released during the acceleration in the supernatant discharge step of all cycles.
| Number | Date | Country | Kind |
|---|---|---|---|
| 2019-119351 | Jun 2019 | JP | national |
| Filing Document | Filing Date | Country | Kind |
|---|---|---|---|
| PCT/JP2020/017761 | 4/24/2020 | WO |
| Publishing Document | Publishing Date | Country | Kind |
|---|---|---|---|
| WO2020/261744 | 12/30/2020 | WO | A |
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| Number | Date | Country | |
|---|---|---|---|
| 20220184639 A1 | Jun 2022 | US |
