JP6295169B2 - Suspension apparatus and suspension method using the same - Google Patents

Description

  The present invention relates to a suspension apparatus and a suspension method using the suspension apparatus, and more particularly to a suspension apparatus using magnetism and a suspension method using the suspension apparatus.

Conventionally, as a suspension device, there is a method for realizing suspension by inserting a stirring bar into a suspension target liquid and vibrating it. However, if it is assumed that the suspension is repeatedly performed for the continuous suspension target liquid, there is a risk of occurrence of contamination via a stirring bar.
Therefore, as the contents of the container to be suspended, put the suspended object and magnetic particles, and move the magnetic particles closer to the magnet from the outside of the container without using a stirrer to realize suspension. There is a technique (for example, refer to Patent Document 1).

JP 2000-254472 A

By using the method as described above, the risk of contamination via a stirrer can be avoided.
However, especially when a highly viscous solution is suspended, the contents lump adheres to the bottom of the container to be suspended in the initial state before suspension, and the contents are quickly washed out and suspended. For this reason, in order to achieve sufficient suspension, the suspension time becomes long.

  Therefore, an object of the present invention is to shorten the time required for sufficient suspension.

  In order to achieve the above object, a suspension device according to the present invention is a suspension device for suspending the contents of a container, and includes a holding unit, a rotation operation unit, and a control unit. ing. The holding unit holds the container. The rotation operation unit rotates the container via the holding unit. The control unit changes the content from the initial state where the container is held by the holding unit to the first state where the contents are located at the upper end of the container, and the contents are located at the lower end of the container from the first state. The rotation operation unit is controlled to change to the second state, the first state period is longer than the second state period, and the first state and the second state are alternated. Control the rotating motion unit to repeat.

  Thereby, the period of a 1st state can be ensured comparatively long, the waiting time until the content adhering to the bottom part of a container falls below can be given stably, and a content is made into a lump. It can be easily dissolved in the solution without stopping. In addition, the contents attached to the bottom of the container are more likely to accumulate than the contents attached near the upper part (ceiling part) of the container, so the control unit controls the period of the first state to be relatively long. This improves the efficiency for melting the contents. As a result, the time required for sufficient suspension can be shortened.

  By configuring the suspension apparatus and the suspension method according to the present invention as described above, the time required for sufficient suspension can be shortened.

1 is a schematic diagram of a suspension device 1 according to an embodiment of the present invention. The flowchart of the suspension method which concerns on embodiment of this invention. The state change explanatory drawing of the centrifuge tube 2 in the rotation suspension which concerns on embodiment of this invention. Explanatory drawing of rotation operation | movement of the centrifuge tube 2 in the rotation suspension which concerns on embodiment of this invention. Explanatory drawing of the rotation center position of the centrifuge tube 2 in the rotation suspension which concerns on embodiment of this invention. Explanatory drawing of the rotation center position of the centrifuge tube 2 in the rotation suspension which concerns on embodiment of this invention. Explanatory drawing of the time change of the rotation angle of the centrifuge tube 2 in the rotation suspension which concerns on embodiment of this invention.

Hereinafter, a suspension apparatus 1 and a suspension method according to an embodiment of the present invention will be described.
(Overview of suspension system)
FIG. 1 is a schematic view of a suspension device 1 according to an embodiment of the present invention, in which FIG. 1 (a) is a top view, FIG. 1 (b) is a front view, and FIG. 1 (c) is a side view.
As shown in FIG. 1, the suspension device 1 mainly includes a holding unit 3, a rotation operation unit 4, a control unit 5, and a magnet unit 6. The suspension device 1 is generally a device that suspends the contents of a container. In this embodiment, the suspension device 1 suspends the solution in the centrifuge tube. In the suspension target solution, a solution containing cells to be suspended and magnetic beads, which are magnetic particles, are mixed, and some of the cells adhere to the magnetic beads.

  The holding | maintenance part 3 hold | maintains the centrifuge tube 2 which is a container. Although various structures can be considered for the rotation operation unit 4, any configuration that is necessary for rotating the centrifuge tube 2 via the holding unit 3 may be used. As a specific configuration example of the holding unit 3, one side of the holding unit 3 is made of rubber, and the other side is made of metal, and the pressure control on the rubber unit is performed by air blowing. Therefore, in the operation of removing the lid of the centrifuge tube 2, the centrifuge tube 2 is configured to have an adsorptive structure so that the centrifuge tube 2 does not move.

In the configuration of the rotation operation unit 4 in FIG. 1C, the rotation center of the centrifuge tube 2 is shown to be the right part of the centrifuge tube 2, but the rotation operation unit 4 includes the centrifuge tube 2. The lower portion, the vicinity of the center of gravity, or the bottom 2c (see FIG. 4 described later) may be configured to be the center of rotation.
The control unit 5 can change the centrifuge tube 2 to various states by controlling the rotation operation unit 4. The magnet unit 6 is disposed in the vicinity of the side portion of the centrifuge tube 2 and is used to draw magnetic beads, which are part of the contents of the centrifuge tube 2, to the side portion of the centrifuge tube 2.

(Overview of suspension method)
Next, a rough flow of the suspension method using the suspension device 1 according to the embodiment of the present invention will be described with reference to FIG.
First, a user installs the centrifuge tube 2 containing the solution to be suspended so that the holding unit 3 of the suspension device 1 can be stably held. In order to perform suspension by a rotating operation, the centrifuge tube 2 is usually one having an upper lid.

  And the control part 5 operates the magnet part 6 so that the magnet part 6 may adjoin the centrifuge tube 2 (S01). Thereby, the magnetic force of the magnet part 6 can be turned ON, and the magnetic beads in the centrifuge tube 2 can be attracted to the side part of the centrifuge tube 2. Here, as a method of switching ON / OFF of the magnetic force, a permanent magnet that always works as a magnetic part may be used as the magnet part 6, and the magnet part 6 may be physically attracted. It is good also as a structure which switches and turns ON / OFF the magnetic force of the magnet part 6 by an electrical switching means.

  Next, the control unit 5 controls the rotation operation unit 4 to rotate the centrifuge tube 2 about several times (S02). Here, to supplement this rotation, the centrifuge tube 2 may be configured to rotate all around, but particularly when the magnetic beads are concentrated and attracted to one side of the centrifuge tube 2, By configuring to rotate about half a turn, the rotation trajectory can be shortened and the suspension speed can be increased.

Thereafter, the control unit 5 leaves the centrifuge tube 2 for about several minutes (S03). At this time, the centrifuge tube 2 is arranged so that the lid of the centrifuge tube 2 is located on the upper side.
Thereafter, after the centrifuge tube 2 is opened, a part of the solution is recovered (S04). Then, the control unit 5 turns off the magnetic force of the magnet unit 6 by operating the magnet unit 6 so that the magnet is separated from the centrifuge tube 2 (S05).

Next, the control unit 5 tilts the centrifuge tube 2 (S06), and the suspension for use in suspension hits the magnetic beads adhering to the side surface in the centrifuge tube 2, and the magnetic beads are removed from the side surface of the container. It is peeled off and added so that it can be recovered in the solution (S07).
Thereafter, after the centrifuge tube 2 is closed, the control unit 5 performs suspension by rotation which is the main embodiment of the present invention (S08). Details will be described separately.

  After suspension of the centrifuge tube 2 in the solution, the control unit 5 operates the magnet unit 6 again so that the magnet unit 6 is close to the centrifuge tube 2 (S09). Thereby, suspended objects, such as a cell adhering to a magnetic bead, can be drawn again. Thereafter, the control unit 5 leaves the centrifuge tube 2 for a few minutes (S10). And after leaving still, the solution of the centrifuge tube 2 is collect | recovered (S11), and suspension is complete | finished.

(Rotating suspension)
Next, the rotational suspension (S09) of FIG. 2 will be described in detail with reference to FIG.
FIG. 3A is a diagram showing the state of the centrifuge tube 2 in the initial state of the rotating suspension, the solution 8 therein, and the magnetic beads 7. As shown in this figure, the centrifuge tube 2 in the initial state is shown in FIG. The centrifuge tube 2 is arranged so that the lid 2a is positioned on the upper side. And inside the centrifuge tube 2, under the influence of the magnetic force of the magnet portion 6, a thick mass of magnetic beads 7 is formed in the side portion 2 b where the magnet portion 6 was close. Further, the magnetic beads 7 are also attached in the vicinity of the lid 2a of the centrifuge tube 2 due to the influence of the rotation in step S02.

  Next, FIG.3 (b) is a figure which shows the 1st state after the control part 5 controls the rotation operation part 4 so that the lid | cover of the centrifuge tube 2 may be located below. As described above, when the control unit 5 controls the rotation operation unit 4, the mass of the magnetic beads 7 attached to the side portion of the centrifuge tube 2 is scraped off due to the shearing force caused by the flow of the solution 8 itself. The magnetic beads 7 are easily dissolved in the solution. Further, at this time, since the solution 8 flows into the vicinity of the lid 2 a of the centrifuge tube 2, which is the lower part in FIG. 3B, the magnetic beads 7 that have been adhered to the vicinity of the lid 2 a of the centrifuge tube 2 until that time are also converted into the solution 8. It becomes easy to melt.

  Next, FIG.3 (c) is a figure which shows the 2nd state after the control part 5 controls the rotation operation part 4 so that the cover 2a of the centrifuge tube 2 may be located in the upper side again. As described above, when the control unit 5 controls the rotation operation unit 4, the shearing force due to the flow of the solution 8 itself is applied, as in FIG. 3B, so that the control unit 5 adheres to the side portion 2 b of the centrifuge tube 2. The lump of the magnetic beads 7 is further scraped off so that the magnetic beads 7 are more easily dissolved in the solution 8. Further, at this time, since the solution 8 flows into the bottom 2c of the centrifuge tube 2 which is the lower part in FIG. 3C, the impact of the solution 8 falling on the lump of the magnetic beads 7 adhering to the bottom 2c of the container is given. Therefore, the magnetic beads 7 attached to the bottom 2c are also easily dissolved in the solution 8.

Then, after repeating the states of FIG. 3B and FIG. 3C, that is, the first state and the second state several times, the initial position as shown in FIG. The control unit 5 controls the rotation operation unit 4 so as to be in the same position.
In addition, the shape of the centrifuge tube 2 in FIG. 3 is a so-called tapered shape in which the bottom 2c is pointed in a conical shape. In such a shape, cells and magnetic beads 7 having a large specific gravity are adsorbed and precipitated as a lump in the vicinity of the bottom, particularly when the suspension is suspended. Therefore, a technique for shortening the suspension time of the solution as in the suspension device according to the present embodiment is particularly useful.

(Rotation operation)
Next, the rotation operation of the centrifuge tube 2 according to the present embodiment will be described with reference to FIG.
As shown in FIG. 4, the centrifuge tube 2 at the initial position is arranged perpendicular to the horizontal line H. Although not shown in the figure, the centrifuge tube 2 is rotated by the rotation operation unit 4 with the bottom 2c as the rotation center axis S1.

  The stationary position 1 where the centrifuge tube 2 is stationary in the first state and the stationary position 2 where the centrifuge tube 2 is stationary in the second state have the same angle with respect to the horizontal line H. The control unit 5 performs software control so that the angle to be formed is switched at predetermined intervals of 30 degrees, 45 degrees, and 60 degrees, for example. In addition, since it becomes difficult to flow a solution stably only to the side surface (especially side part 2b) of the centrifuge tube 2 when it becomes close to 90 degree | times, in order to make use of the force which a solution flows, a solution Although it varies depending on various conditions such as viscosity, concentration or magnetic beads, a setting of about 60 degrees is desirable. The end position of the centrifuge tube 2 is the same as the initial position.

  The adjustment of the rotation angle can be realized by software control of the control unit 5 as described above. However, by separately providing a partition plate (not shown), the centrifuge tube 2 can be mechanically rotated. By mounting, it can be suppressed to a predetermined range, and further stabilization of suspension processing can be realized. As an implementation example other than providing a partition plate, by using a positioning sensor (not shown) for continuously observing whether or not a jig such as the rotary operation unit 4 has passed a predetermined position, the centrifuge tube 2 It is good also as a structure which controls a rotation angle. Moreover, it is good also as a structure which controls a rotation angle according to a suspension condition by image | photographing a suspension condition with a camera etc., and performing an analysis using an algorithm.

(Modification of the position of the rotation center)
Next, the position of the rotation center of the centrifuge tube 2 according to the embodiment of the present invention will be described with reference to FIGS. 5A and 5B.
As shown in FIG. 5A, the center point S2 of the rotation of the centrifuge tube 2 by the rotation operation unit 4 may be at a position different from the center position S1 of the centrifuge tube 2 held by the holding unit 3. This is to increase the centrifugal force to the solution 8 and the magnetic beads 7 inside the centrifuge tube 2 by increasing the distance from the rotation center point S2 of the centrifuge tube 2 to the end of the centrifuge tube 2. By applying this centrifugal force, it is possible to easily separate the magnetic beads 7 that easily collect in the bottom 2c from the bottom 2c. Further, since the kinetic energy of the solution 8 and the magnetic beads 7 is increased by the centrifugal force, the impact force that strikes the magnetic beads 7 near the bottom 2c and the lid 2a of the centrifuge tube 2 at the stationary position 1 and the stationary position 2, and the solution The impact force that 8 breaks the magnetic beads 7 can be increased, and the efficiency for melting the magnetic beads 7 is improved. As a result, the time required for sufficient suspension can be shortened.

As shown in FIG. 5A, the centrifuge tube 2 may be arranged so that the rotation axis S2 of the rotation of the centrifuge tube 2 is positioned on the extension line from the lid 2a to the bottom 2c of the centrifuge tube 2, As described above, the centrifuge tube 2 may be arranged so as to be in contact with a line connecting one point of the bottom 2c of the centrifuge tube 2 and the rotation axis S3 of the rotation of the centrifuge tube 2 only at one point of the bottom 2c.
(Time change of rotation angle)
Next, the time change of the rotation angle of the centrifuge tube 2 according to the embodiment of the present invention will be described with reference to FIG. The horizontal axis in FIG. 6 is time, and the vertical axis is the rotation angle of the centrifuge tube 2. The centrifuge tube 2 is appended in the drawing so that the rotation angle of the centrifuge tube 2 can be easily seen.

At time t0, the centrifuge tube 2 is in the initial state. Thereafter, the state of the centrifuge tube 2 becomes the first state due to the rotation in the forward direction (see arrow J in FIG. 4), and stops at time t1. The period from time t1 to time t2 is the first stationary period.
Thereafter, the centrifuge tube 2 rotates in the reverse direction (see the arrow K in FIG. 4) so as to be in the second state, and stops at time t3. The period from time t3 to time t4 is the second stationary period. Thereafter, the centrifuge tube 2 rotates again in the forward direction (see arrow J in FIG. 4) so that the state of the centrifuge tube 2 becomes the first state, and stops at time t5. In this way, the first stationary period and the second stationary period are alternately repeated. The period of the first state is longer than the period of the second state, and the first stationary period is longer than the second stationary period. By securing the first state and the first stationary period, in particular, when the tapered centrifuge tube 2 is employed, there is a waiting time until the magnetic beads 7 that tend to collect on the bottom 2c of the centrifuge tube 2 fall downward. As a result, the magnetic beads 7 can be easily dissolved in the solution 8 without being retained as a lump. Then, by ensuring the second state and the second stationary period, the magnetic beads 7 attached to the vicinity of the lid 2a of the centrifuge tube 2 can be easily dissolved in the solution 8, and all of the solution 8 to be suspended is removed. It can be moved downward.

  Further, only the characteristic part including the configuration of the suspension device 1 will be described in detail. The controller 5 starts from the initial state in which the centrifuge tube 2 is held by the holding unit 3, and the contents are transferred to the upper end of the centrifuge tube 2. The rotation operation unit 4 is controlled so that the first state is changed, and the first state is changed to the second state where the contents are located at the lower end of the centrifuge tube 2. Furthermore, the control unit 5 controls the rotation operation unit 4 so that the period of the first state becomes longer than the period of the second state, and the first state and the second state are alternately repeated. To do.

  By comprising in this way, the period of a 1st state can be ensured comparatively long, and the waiting time until the magnetic bead 7 adhering to the bottom part 2c of the centrifuge tube 2 falls below is given stably. The magnetic beads 7 can be easily dissolved in the solution 8 without being retained as a lump. Since the magnetic beads 7 adhering to the bottom 2c of the centrifuge tube 2 are more likely to accumulate than the magnetic beads 7 adhering to the vicinity of the lid 2a, the control unit 5 controls the period of the first state to be relatively long. The efficiency for melting the magnetic beads 7 is improved. As a result, the time required for sufficient suspension can be shortened.

  Here, supplementing the change from the initial state to the first state and the change from the first state to the second state, as described above with reference to FIG. 4, the centrifuge tube 2 is in the initial state. The upper end 2d is arranged perpendicularly to the horizontal line H, the upper end 2d is inclined about 60 degrees downward with respect to the horizontal line in the first state, and the upper end 2d is arranged with respect to the horizontal line in the second state. It is tilted upward by about 60 degrees. However, the present invention is not limited to these arrangement settings, and the centrifuge tube 2 is inclined and arranged so that the contents are located at the upper end of the centrifuge tube 2 in the first state, and the contents are stored in the centrifuge tube 2 in the second state. If the centrifuge tube 2 is disposed so as to be positioned at the lower end, the efficiency for melting the magnetic beads can be increased as described above.

Then, as shown in FIG. 6, the control unit 5 has a first stationary period in which the rotation of the centrifuge tube 2 is stationary in the first state, and the rotation of the centrifuge tube 2 is performed in the second state. It is desirable to control the rotation operation unit 4 so as to have a second stationary period so as to be stopped, and to make the first stationary period longer than the second stationary period.
In FIG. 6, the rotation speed of the rotation operation unit 4 is controlled so that the waveform of the time displacement of the rotation angle is substantially trapezoidal, such as constant in the forward direction → zero (stationary) → constant in the reverse direction. Although controlled by the unit 5, the shape is not limited to this shape, and the configuration may be such that the control unit 5 controls the waveform of the temporal displacement of the rotation angle to be a sine waveform. Furthermore, in FIG. 6, the slope of the straight line indicating the temporal displacement of the rotation angle in the period t2 to t3 from the first stationary period to the second stationary period, that is, the constant speed in the forward direction, That is, the slope of the straight line indicating the time displacement of the rotation angle in the period t4 to t5 from the second stationary period to the first stationary period is steeper, and the period t2 to t3 is set shorter than the period t4 to t5. Has been. Thus, when changing from the first quiescent period to the second quiescent period, the period is set to be relatively short, so that the magnetic beads adhering to the side surface of the centrifuge tube 2 are washed away vigorously. Furthermore, the magnetic beads accumulated at the bottom of the centrifuge tube 2 can be crushed by the impact force caused by the dropping of the solution. Further, when changing from the second stationary period to the first stationary period, the period is set to be relatively long, so that the magnetic beads 7 accumulated at the bottom of the centrifuge tube 2 are reduced. Thus, it is possible to prevent only the solution from flowing and the magnetic beads 7 from staying near the bottom.

  When the control unit 5 performs control so as to repeat the period of the first state and the period of the second state, or the first stationary period and the second stationary period, each period is repeated. You may change so that the length of may be shortened as a whole. Furthermore, the control unit 5 determines the number of repetitions of the first stationary period and the second stationary period, or the first stationary period and the second stationary period, based on the results measured in the experiment. In addition, a predetermined configuration may be used. This number of times varies depending on various conditions such as the type of suspension object and the amount of liquid.

  Although not shown, the suspension device 1 may be configured to further include a sensor that detects a partial state of the centrifuge tube 2. In this case, the control unit 5 changes the control of the rotation operation unit 4 based on the detection result of the sensor. That is, after the magnetic beads 7 are no longer detected by continuously detecting the state of the part where the magnetic beads 7 are likely to collect, such as the bottom, side, or near the lid of the centrifuge tube, the control unit By configuring 5 to stop the control of the rotation operation unit 4, more stable suspension can be realized even if the number of repetitions in suspension is not set in advance as described above. As a type of sensor, a sensor having a light emitting function of reflecting the magnetic beads without transmitting light may be used.

  Here, as an example relating to a more specific sensor arrangement, the sensor is fixedly arranged in a place not affected by the rotation operation, and the centrifuge tube 2 in the first stationary state and the second stationary state described above, for example. It may be configured to detect only a part of the. Moreover, as another example regarding the arrangement of the sensor, the sensor is connected to the rotation operation unit 4 and rotated together with the rotation of the centrifuge tube 2 by the rotation operation unit 4, while the centrifuge tube 2 is rotating. You may arrange | position so that the detection of a part of location may be continued. Moreover, you may use an imaging means instead of a sensor.

  As mentioned above, although one Embodiment of this invention was described, this invention is not limited to the said embodiment, A various change is possible in the range which does not deviate from the summary of invention.

  Since the suspension device according to the embodiment of the present invention is a device having a short time required for sufficient suspension, the suspension target is difficult to dissolve in a solution such as a cell, such as a cell culture device. And is particularly useful in an apparatus using magnetic particles or the like for suspension.

DESCRIPTION OF SYMBOLS 1 Suspension apparatus 2 Centrifuge tube 2a Lid 2b Side part 2c Bottom part 2d Upper end part 3 Holding part 4 Rotation operation part 5 Control part 6 Magnet part 7 Magnetic bead 8 Solution

Claims (10)

A suspension device for suspending the contents of a container,
A holding part for holding the container;
A rotation operation unit for rotating the container via the holding unit;
The initial state in which the container is held by the holding unit is changed to a first state in which the contents are located at the upper end of the container, and the contents are moved from the first state to the lower end of the container. The rotation operation unit is controlled so as to change to the second state, and the period of the first state is longer than the period of the second state, and the first state A control unit for controlling the rotation operation unit so as to alternately repeat the second state;
A suspension apparatus comprising:   In the control unit, the first stationary period in which the rotation of the container is stationary in the first state is longer than the second stationary period in which the rotation of the container is stationary in the second state. The suspension device according to claim 1, wherein the rotation operation unit is controlled.   The suspension device according to claim 1 or 2, wherein the container is a centrifuge tube having a bottom portion that is pointed in a columnar shape.   The magnet part which is arrange | positioned near the side part of the said container, and is used in order to draw the magnetic particle which is a part of the contents of the said container to the side part of the said container is further provided. The suspension apparatus according to 1. The container is a centrifuge tube having a lid at the upper end,
The controller is
In the initial state, arrange the centrifuge tube so that the lid is located on the upper side,
In the first state, the rotation operation unit is controlled so that the lid is positioned on the lower side,
In the second state, the rotation operation unit is controlled so that the lid is positioned on the upper side.
The suspension device according to any one of claims 1 to 4.   The suspension device according to any one of claims 1 to 5, further comprising a partition plate that restricts the rotation operation of the rotation operation unit within a predetermined range. A sensor for detecting a state of a part of the container;
The suspension device according to any one of claims 1 to 6, wherein the control unit changes control of the rotation operation unit based on a detection result of the sensor.   The suspension device according to claim 7, wherein the sensor rotates together with the rotation of the container by the rotation operation unit, and continues to detect a part of the same portion of the container while the container is rotating.   The suspension device according to any one of claims 1 to 8, wherein a center point of rotation of the container by the rotation operation unit is different from a center position of the container held by the holding unit. A suspension device for suspending the contents of a container, a holding unit for holding the container, a rotation operation unit for rotating the container via the holding unit, and a control unit for controlling the rotation operation unit; A suspension method using the suspension device comprising:
The control unit controls the rotation operation unit so that the content is changed from an initial state in which the container is held by the holding unit to a first state in which the contents are located at an upper end of the container;
The control unit controls the rotation operation unit so that the content of the container changes from the first state to a second state located at the lower end of the container, and the period of the first state Controlling the period of the second state to be shorter than
The control unit controlling the rotation operation unit so as to alternately repeat the first state and the second state;
A suspension method comprising:

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