JPH0135699B2 - - Google Patents

Description

DETAILED DESCRIPTION OF THE INVENTION [Field of Application of the Invention] The present invention relates to a suspension processing device, and particularly for suspending particles in a container when processing a solution containing particles with sedimentation properties in a container. The present invention relates to a suspension processing device suitable for.

[Background of the invention]

A blood cell washing device that uses centrifugation is known as one of the devices that handle sedimentary particles. As shown in Figure 1, the conventional washing process of a washing centrifuge that washes blood cells (biological particles) is as follows: First, a washing liquid is injected into a test tube containing a sample using a dispenser, and then, After rotating the centrifuge to sediment particles such as blood cells, the supernatant liquid was discarded.

This conventional cleaning method has no problems when cleaning blood cells such as relatively free-flowing red blood cells or polymer particles such as latex, but when cleaning blood cells with relatively high viscosity such as lymphocytes, ,
It is not suitable for cleaning biological particles that are easily destroyed by high-speed injection of cleaning fluid. This is because relatively free-flowing blood cells such as red blood cells are stirred almost uniformly by the ejection force when injecting the washing liquid, but blood cells with relatively high viscosity such as lymphocytes are mixed almost uniformly. The discharge force for injecting the cleaning liquid alone will not stir and create a uniform suspension, and increasing the discharge force for injecting the cleaning liquid will improve the stirring effect, but
This is because if this is done, the cleaning liquid may spill out of the test tube or the sample to be cleaned (biological particles) may be destroyed.

Particularly in the case of washing lymphocytes, etc., strong stirring will destroy the platelets mixed in the lymphocytes. When platelets are destroyed, lymphocytes form pellet-like clumps, and individual lymphocytes are not washed at all.

Conventional blood cell washing centrifuges have the above-mentioned problems, and when a blood cell washing centrifuge is not used, the operator has to repeatedly inhale and dispense the washing liquid with a pipette to uniformly suspend the sedimented blood cells. After
The extremely troublesome operations of performing centrifugation and discarding the supernatant solution require a great deal of effort.

However, in recent years, lymphocytes have become an indispensable research subject in the field of immunology, and the number of processed specimens has increased significantly.

[Purpose of the invention]

An object of the present invention is to provide a suspension processing device that has a simple configuration and can enhance the suspension effect of sedimentary particles.

[Summary of the invention]

The present invention provides a rotating body in which a plurality of containers containing a liquid containing particles having sedimentation properties are arranged on a circumference, and the rotating body is given rotation in one direction according to the process, and rotation in forward and reverse directions. A single drive device that alternately applies the power, and controls this drive device to cause the rotating body to repeat forward and reverse rotations in small steps to perform a stirring operation of the liquid in the plurality of containers, and then the rotating body The present invention is characterized in that a control device is provided for controlling the drive device so that the drive device rotates in one direction.

[Embodiments of the invention]

As an example of handling a container containing a liquid containing sedimentary particles, an example will be described in which the present invention is applied to a blood cell washing centrifuge. The configuration and operation of an embodiment of the present invention will be described with reference to FIGS. 2 to 6.

FIG. 2 shows a schematic configuration of an embodiment of the present invention. In FIG. 2, a plurality of centrifugal test tubes 2 each containing a liquid containing sedimentary particles, such as a lymphocyte suspension, are arranged on the circumference of a rotor 3. When processing lymphocytes for immunological research, it is necessary to centrifuge the lymphocytes in peripheral blood cells to separate them from other components. However, since plasma and other blood cells are mixed in the centrifuged lymphocyte layer, unnecessary substances must be removed by washing. Test tube 2 contains a liquid containing lymphocytes before washing.

The plurality of test tubes 2 are each attached to a test tube holder 1 swingably attached to the outer circumferential direction of the rotor 3. An iron piece 4 is attached below the holder 1, so that when the electromagnet 5 is energized, the lower part of the holder 1 is attracted to the electromagnet 5 side via the iron piece 4. 6 is rotor 3
Motor 6 is a centrifugal rotation pulse motor that rotates
The rotation speed, rotation time, and forward/reverse operation of the rotation are controlled by the motor controller 7. In addition,
The motor 6 is fixed to a pedestal 10 via a seismic isolation spring 8 and a mounting bracket 9.

On the other hand, a washing liquid (physiological saline, phosphate buffer, etc.) 11 is sucked from a bottle 13 by a washing liquid dispensing syringe 12, passed through a flow path switching valve 14, and injected into the test tube 2 from a washing liquid injection nozzle 15. Ru.
The flow path switching valve 14 switches the flow path when dispensing the cleaning liquid.
This is performed by the interlocking operation of the motor 16 for vertically driving the syringe 12.

After the washing centrifugation, the waste liquid discarded as a supernatant is collected in the waste liquid tank 17. 18 is the case, 19
is an opening/closing lid.

Next, the cleaning process in this example will be explained. FIG. 3 is an explanatory diagram of the cleaning process. First, test tube 2 containing about 0.5 to 1 ml of lymphocyte suspension solution.
is attached to the swing-type centrifugal test tube holder 1, and as shown in FIG. 4, the rotor 3 is repeatedly rotated forward and reverse in small steps by controlling the forward and reverse rotation of the centrifugal rotation motor 6 by the motor controller 7. As a result, each test tube 2 swings and the liquid is stirred by vibration, and the lymphocytes are floated and uniformly suspended in the liquid. Next, after this state, the rotor is stopped and cleaning liquid is injected. At this time, since the lymphocyte suspension has already become suspended and homogeneous, stirring is reliably performed by the ejection force generated by the injection of the washing liquid. The lymphocytes are therefore completely disentangled and suspended in the washing solution. In this case, resuspension and agitation cannot be achieved completely by simply injecting a washing solution directly into the lymphocyte suspension as in the conventional method.

Next, after the cleaning liquid is injected, the rotor 3 is centrifugally rotated in one direction at high speed. The state of centrifugal rotation at this time is shown in FIG. Due to the centrifugal rotation of the rotor 3, the lymphocytes in the test tube 2 are sedimented by centrifugal force and settle to the bottom of the test tube 2. After centrifugation, the rotor is temporarily stopped and the supernatant liquid is discarded (decanted). FIG. 6 is a diagram showing the state of decantation. At this time, the electromagnet 5 of the centrifugal test tube holder 1 is energized, so the iron piece 4 is attracted to the electromagnet 5, and the bottom part is on the side of the rotating shaft of the rotor 3. attracted to test tube 2
When the opening of the rotor 3 is turned outward and the rotor 3 is rotated in this state, the supernatant liquid flows out and is discarded. At this time, the rotation time is controlled so that about 0.5 to 1 ml of cleaning liquid remains. After the supernatant liquid is discarded, stir by rotating forward and backward in the same way as above.
Thereafter, the cleaning process described above is repeated.

The above-described cleaning process is automatically performed by controlling the forward rotation, reverse rotation, rotation time, and rotation speed of the centrifugal rotation motor 6.

Next, the relationship between stirring and cleaning effect will be explained. If the stirring is insufficient and the lymphocytes are localized at the bottom of the test tube 2, foreign substances mixed in the lymphocytes will not come out into the washing solution. Therefore, if agitation (resuspension) is insufficient, no matter how many times the cleaning is repeated,
Lymphocytes cannot be washed well. Also, when decanting the washing solution, leaving a small amount of the supernatant rather than completely discarding it may reduce the washing effect, but if the supernatant is completely discarded, the next This is not preferable because it causes insufficient stirring in the step. Furthermore, if the washing solution is completely discarded, there is a risk that a portion of the lymphocytes will also be thrown away, leading to a decrease in the recovery rate in the washing step.

Therefore, the cleaning process as in this example is effective in improving the cleaning effect and at the same time preventing the recovery rate from decreasing in the cleaning process.

According to the embodiment of the present invention described above, the following advantages can also be obtained.

(b) Lymphocytes can be sufficiently stirred, resuspended completely, and lymphocytes can be thoroughly washed, which was previously impossible.

(b) Automatic washing of lymphocytes is possible, which saves labor.

(c) It is possible to improve the recovery rate in the washing process, and specifically, a recovery rate of 80% or more can be obtained.

(d) Platelets mixed in lymphocytes are not destroyed during washing, and lumps of lymphocytes during the washing process can be eliminated, and a homogeneous lymphocyte suspension can be provided as an analysis sample.

(E) A series of operations of stirring, centrifugation, and decantation can be performed by controlling a single motor without adding any other stirring mechanism.

Note that it is conceivable to attach a vibrating mechanism such as a pibrator to the rotor 3 to vibrate the test tube 2, but in this case, another mechanism would be added.

In the above embodiment, an example of application to a blood cell washing centrifuge was described, but the suspension stirring method according to the present invention can also be applied to an immunoassay device using particles such as latex.

〔Effect of the invention〕

As explained above, according to the present invention, a liquid containing sedimentary particles in a container can be stirred with a simple configuration, so that sedimentary particles can be uniformly suspended in the liquid, especially when the particles are relatively viscous. Even when processing particles with a high particle content or particles that are easily destroyed, they can be suspended well by forward and reverse rotation stirring.

[Brief explanation of drawings]

FIG. 1 is a diagram explaining the washing process in a conventional blood cell washing centrifuge, FIG. 2 is a diagram showing a schematic configuration of a blood cell washing centrifuge which is an embodiment of the present invention, and FIG. 3 is an implementation of the method shown in FIG. 2. A diagram illustrating a cleaning process in an example,
Figure 4 shows the stirring state of the rotor in Figure 2 by forward and reverse rotation, Figure 5 shows the centrifugal operation of the rotor in Figure 2, and Figure 6 shows how the supernatant liquid is discarded from the test tube. FIG. 1... Test tube holder, 2... Centrifuge test tube,
3... Rotor, 4... Iron piece, 5... Electromagnet, 6...
...Centrifugal rotating motor, 7...Motor controller, 11...
...Cleaning liquid, 12...Cleaning liquid dispensing syringe, 15...
...Cleaning liquid injection nozzle.

Claims (1)

[Claims] 1. A rotating body in which a plurality of containers containing a liquid containing particles having sedimentation properties are arranged in a circumferential manner, and this rotating body is given rotation in one direction and alternately with forward and reverse rotation depending on the process. A single motor and the operation of the motor are controlled to cause the rotating body to repeat forward and reverse rotations in small steps to stir the liquid in the plurality of containers, and then the rotating body moves in one direction. 1. A suspension processing device, comprising: a control device for controlling the motor so as to rotate the motor.