JPH0636732B2 - Cell disruptor - Google Patents

Abstract

PURPOSE:To enable the disintegration of cells of a number of specimens at a time by providing a rotary shaft with an inclined shaft part, fitting a ring to the inclined shaft in a state to keep the ring from rotation together with the rotary shaft and attaching closed vessels to the outer circumference of the ring. CONSTITUTION:A ring member 15 freely rotatable relative to an inclined shaft 8 clamped and fixed to a rotary shaft 6 is fit to the outer circumference of the inclined shaft 8 via a pair of bearings 13. A closed vessel 20 containing various kinds of cells to be disintegrated and micro-beads of 0.1-0.5mm in diameter is attached between the holder pieces 18a, 18b on the outer circumference of the ring member 15. A number of protrusion 17 are radially protruded from the outer circumference of the ring member 15 at a prescribed pitch in circumferential direction and a tension spring 23 is extended between a spring hook 22 of the protrusion and a mount 3 to prevent the rotation of the ring member 15 together with the rotary shaft 6. The rotary shaft 6 is rotated at 1,200-1,800rpm with a motor 5 to effect the swinging motion of the ring member to both sides in the direction of shaft center. The closed vessel 20 is oscillated in the form of figure 8 according to the high-speed rotation of the rotary shaft 6 to disintegrate the cells in the closed vessel 20 with the micro-beads.

Description

DETAILED DESCRIPTION OF THE INVENTION (Field of Industrial Application) The present invention relates to a cell disruption device for disrupting cells and tissues.

(Prior art) As a cell crushing device for screening substance-producing bacteria such as yeast and bacteria and producing cells, various crushing methods such as crushing with a high-speed rotary knife, crushing with ultrasonic waves, crushing with pressure, etc. are adopted. Among them, the cells to be crushed and the micro beads made of glass or ceramics are housed in a sealed container, and this sealed container is vibrated at high speed, and the collision of the micro beads and the cells is repeated to move the cells. A crushing method is also known.

(Problems to be Solved by the Invention) However, the conventional micro-bead impact crushing device is configured so that a single closed container can be set, which is very difficult when crushing cells of a large number of samples. There was a problem that it took time and was inefficient.

Therefore, a configuration in which a large number of container holders are arranged on a vibrating body so that a large number of containers can be set is conceivable. However, in order to cause such a vibrating body to perform an 8-shaped vibration, which is the optimum vibration mode. Then, there is a problem that the configuration is complicated and large.

In view of the above conventional problems, it is an object of the present invention to provide a cell disruption device capable of performing cell disruption of a large number of samples at a time, and having a simple configuration and an optimal vibration mode into a figure-eight shape. And

(Means for Solving the Problem) In order to achieve the above-mentioned object, the cell disruption device of the present invention is provided with an inclined shaft portion whose center line is inclined with respect to the axis of the rotational shaft that can be rotationally driven. An annular body is rotatably fitted to the shaft so that its center line coincides with the shaft, and a spring that elastically restrains the rotation of the annular body is attached to the annular body. And a plurality of holders for holding a closed container containing beads are arranged in the circumferential direction.

(Operation) According to the cell disruption device of the present invention, the rotation of the ring-shaped body is such that the ring-shaped body is relatively rotatably externally fitted to the inclined shaft portion provided on the rotation shaft and the ring-shaped body does not rotate together with the rotation shaft. The structure is elastically constrained by a spring, and the center line of the inclined shaft portion and thus the annular body is inclined with respect to the axis of the rotating shaft. Therefore, the outer periphery of the annular body is rotated every time the rotating shaft makes one rotation. Each point of moves in a figure eight shape. Therefore, when the rotating body is rotated at a high speed, the contents (the crushed object and the beads) of the closed container arranged through the holder on the outer periphery of the annular body are vibrated in the shape of a figure eight, and the crushed object is Effective collision with beads,
It is crushed efficiently in a short time due to friction.

In addition, since many holders are arranged on the outer circumference of the annular body, it is possible to disrupt many cells at one time.

(Embodiment) An embodiment of the present invention will be described below with reference to FIGS. 1 to 4.

Reference numeral 1 is a base, on which a pedestal 3 is installed via a vibration-proof rubber 2. A pair of bearing units 4a and 4b and a motor 5 are arranged on the pedestal 3 such that their axes are aligned. A pair of bearing units 4a,
4b is a bearing stand 3 which is arranged at an appropriate distance from each other.
It is installed on a and 3b, and rotatably supports both ends of the rotary shaft 6. One end 6a of the rotary shaft 6 is connected to the output shaft 5a of the motor 5 via a coupling 7. Between the bearing units 4a and 4b of the rotary shaft 6, an inclined shaft body 8 having an inclined hole 8a whose center line is inclined with respect to the central axis is fitted and fitted to a key (not shown). Are connected so that they cannot rotate relative to each other. The inclined shaft body 8 is formed on the outer peripheral surface of the rotary shaft 6 and the flange portion 10 projecting on the outer peripheral surface of the rotary shaft 6 via the inclined spacer rings 9a and 9b arranged in contact with both end surfaces thereof. A fixed nut 12 screwed into the screw portion 11 is used to narrowly fix the shaft in the axial direction, and constitutes an inclined shaft portion that rotates integrally with the rotating shaft 6.

An annular body 15 is fitted onto the outer periphery of the inclined shaft body 8 via a pair of bearings 13 so as to be relatively rotatable. Reference numeral 14 is a bearing fixing nut on the side of the inclined shaft body 8, and 16 is a bearing fixing nut on the side of the annular body 15. On the outer circumference of the annular body 15, a large number (12 in the illustrated example) of projections 17 are radially provided at a predetermined pitch in the circumferential direction. Holder pieces 18a and 18b made of elastic plates are attached to both end surfaces of the protrusion 17 by screws 19 so as to project outward in the radial direction and face each other, and a tubular shape is provided between the holder pieces 18a and 18b. The closed container 20 is detachably held.

A connecting portion 21 is formed between two adjacent protrusions 17, 17 and a spring hook 22 is attached by flattening the outer peripheral surface thereof, and a tension spring 23 is provided between the spring hook 22 and the gantry 3. It is interposed and prevents rotation of the annular body 15 so as not to rotate together with the rotating shaft 6. Further, a stopper shaft 24 passing through the central portion of the space formed between the one protrusion 17 connected to each other by the connecting portion 21 and the protrusion 17 adjacent to one side thereof is provided, and both ends thereof are provided. Part is bearing stand 3
It is supported by a and 3b.

Next, the operation will be described. A variety of cells to be crushed as samples and microbeads made of glass or ceramics are respectively housed in a tubular closed container 20, and the closed container 20
Is held between the holder pieces 18a and 18b on the outer periphery of the annular body 15. The fine beads are ceramic beads such as glass beads or zirconia having a diameter of 0.1 to 0.5 mm,
For example, it is selected and used according to the purpose of disruption such as disruption of microorganisms or disruption of eukaryotic cells.

When the mounting of the closed container 20 is completed, the motor 5 is driven to rotate the rotating shaft 6 at high speed, for example, 1200 to 1800 rpm. Then, the annular body 15 is fitted onto the outer periphery of the inclined shaft body 8 which rotates integrally with the rotating shaft 6 so as to be relatively rotatable, and the co-rotation is prevented by the tension spring 23, as shown in FIG. Every time the rotary shaft 6 rotates once, the annular body 6 oscillates to both sides in the axial direction.

At this time, an arbitrary point on the outer circumference of the annular body 15 is shown in FIG.
As shown in (a) and (b), it moves in a figure eight shape.
That is, as shown by the solid line in FIG. 4 (a), the behavior of the point a on the outer periphery of the annular body 15 at that time is taken as a reference position with the annular body 15 tilted in the vertical direction of the paper surface. Then, when the rotary shaft 6 rotates 90 ° in the direction of the arrow from the solid line state, the annular body 15 moves slightly in the front and back directions of the paper surface, and shifts to the state shown by the phantom line, corresponding to point a during that time. The moved position moves to the point c via the arcuate path b. Next, when the rotary shaft 6 further rotates 90 °, the annular body 15 shifts to the state shown by the solid line in FIG.
The position corresponding to the point returns from the point c to the original point a via the arc-shaped path d. When the rotary shaft 6 further rotates 90 °,
The annular body 15 shifts to a state shown by an imaginary line, the position corresponding to the point a moves to the point f via the arcuate path e, and the rotating shaft 6 rotates 90 ° to the original rotating position. Then, the point f corresponding to the point a returns to the original point a via the arcuate path g. The arcuate path b,
The loci of d, e, and g are determined by the balance between the elastic restraining force of the tension spring 23 and the frictional resistance of the inclined shaft body 8 with respect to the annular body 6, and when the elastic restraining force is large, the locus is It will approach a straight line.

Therefore, the closed container 20 held on the outer periphery of the annular body 15 is vibrated in the shape of a figure 8 as the rotating shaft 6 rotates at high speed, and as a result, the microbeads are crushed on the cells to be crushed in the closed container 20. The collision is effective, and the collision rapidly and uniformly breaks the cells. In this way, cell crushing of a large number of samples contained in a large number of closed containers 20 can be performed at once.

The annular body 15 has its outer peripheral position elastically pulled by a tension spring 23 to prevent its rotation while allowing the above-described 8-shaped displacement operation. In some areas, abnormal vibration due to resonance may occur, but in that case, since the protrusion 17 contacts the stopper shaft 24, the abnormal vibration is reliably prevented.

In the above-described embodiment, the example in which the single annular body 15 is arranged on the rotating shaft 6 is shown, but as shown in FIG. 5, a plurality of annular bodies 15 having the same structure are arranged in parallel on the rotating shaft 6. It is also possible to perform cell disruption of a larger sample at a time.

A box 50 that can be opened and closed as shown in phantom in FIG.
It is preferable that the entire annular body 15 be covered with and the cold air 51 be sent to the annular body 15 to cool the inside of the closed container 20 that is heated by vibration during crushing. The box 50 also functions as a safety box that prevents the closed container 20 from scattering even if it is separated from the holder pieces 18a and 18b.

(Effects of the Invention) According to the cell disruption device of the present invention, a large number of samples can be disrupted at one time by holding a closed container in a large number of holders arranged around a ring-shaped body, and the cell is provided on a rotating shaft. Since each closed container is held on the outer periphery of the annular body fitted to the inclined shaft portion so as not to rotate together by the restraint by the spring, each closed container moves in a figure 8 shape each time the rotating shaft makes one rotation. By rotating the rotary shaft at a high speed, it is possible to vibrate each closed container in an optimum vibrating form of a figure eight shape. In addition, the rotary shaft is provided with an inclined shaft portion, and the annular body is externally fitted to the rotary shaft, so that a simple structure can be obtained at a low cost.

[Brief description of drawings]

1 to 4 show an embodiment of the present invention, FIG. 1 is a partial sectional front view, FIG. 2 is a side view, and FIG. 3 is a front view of a main part showing an operating state, FIG. (a), (b) is explanatory drawing of the vibration form in the outer periphery of an annular body, and FIG. 5 is a front view of the principal part of other Example of this invention. 6 ... Rotating shaft 8 ... Inclined shaft body 15 ... Annular body 18a, 18b ... Holder piece 20 ... Closed container 23 ... Tension spring

Claims (1)

[Claims] 1. A rotary shaft which can be rotationally driven is provided with an inclined shaft portion whose center line is inclined with respect to the axis thereof, and an annular body is fitted relatively rotatably outside so that the center line coincides with the inclined shaft portion. At the same time, a spring that elastically restrains the rotation is attached to this annular body, and a large number of holders for holding a closed container containing the crushed object and beads are arranged in the circumferential direction on the outer periphery of the annular body. A cell disruption device characterized by: