JP2005131475A - Crusher - Google Patents

Abstract

<P>PROBLEM TO BE SOLVED: To provide a cell crusher constituted so as to freeze tissue pieces or the like including the liver, heart and smooth muscle of an experimental animal or the like to crush the same simply and rapidly and enhanced in crushing efficiency and the recovery of a useful substance. <P>SOLUTION: In this cell crusher constituted so that a screw shaft driven by a motor is supported in a housing cylinder having a supply port in a rotatable manner and the rotary blade directly connected to the screw shaft is attached to the discharge side of the housing cylinder, a means for detecting overload is provided to the motor and the motor is instantaneously reversed after the detection of overload to efficiently crush the frozen tissue pieces of a living body. <P>COPYRIGHT: (C)2005,JPO&NCIPI

Description

  The present invention relates to a crushing device for the purpose of taking out enzymes, DNA, RNA, proteins, anticancer substances, intracellular micro-tissue, etc., which are useful substances in cells or tissues by crushing living tissue cells, The present invention relates to a crushing apparatus that is easy to handle, can process a large amount of ecological tissue quickly and safely, and has high crushing efficiency and a high recovery rate of useful substances.

  Conventional cell crushing devices include, for example, those disclosed in Japanese Published Unexamined Patent Application No. 2-504360 or those shown in FIG. This is also called a homogenizer, and is a method of crushing with a screw-shaped blade rotating at high speed, or a method of crushing between a rotating inner blade and a fixed outer blade.

In such a crushing apparatus, it is suitable for crushing a small soft piece of tissue that has been cut into small pieces. However, for a tissue containing smooth muscle such as an animal heart or a hard tissue such as bone, only a rotating blade is used. In this method, the crushing efficiency is low, and it takes time to crush the tissue pieces to a size that allows protein, DNA, RNA, etc. to be collected. The tissue is caught, the crushing capacity is remarkably reduced, and the processing capacity is not satisfactory. Furthermore, in the cell crushing apparatus, since the rotating blade for crushing rotates at a high speed (for example, 27,000 revolutions per minute or more), when crushing the cells, the rotating blade is applied to the tissue piece many times at a high speed. The frictional heat generated by the friction between the rotating blade and the tissue causes the heat-sensitive material such as protein, DNA, RNA, etc. to be extracted from the cells to be altered, and the protein, DNA, Since substances such as RNA are easily damaged, the recovery rate is significantly reduced.
Furthermore, the tissue may be stored frozen, and there is a desire to grind the tissue that has been frozen.

Publication 2-504360

  The present invention provides a crushing apparatus that improves the above problems, is easy to handle, can process a large amount of biological tissue cells quickly and safely, can crush a large amount of samples efficiently, and has a high recovery rate of useful substances. The purpose is to do.

  In order to solve the above problems, a crushing apparatus according to the present invention includes a housing, a motor disposed in the housing, a storage cylinder having a supply port and a discharge port, and a screw driven by the motor in the storage cylinder. The shaft is rotatably supported, a rotary blade that rotates together with the screw shaft is attached to the discharge side of the storage cylinder, and a fixed blade is attached to the storage cylinder side facing the rotary blade. In the crushing apparatus, the motor is provided with means for detecting an overload, and the motor is reversed by an output signal from the means for detecting the load.

  The means for detecting the overload of the crushing device is achieved by detecting the rotational speed of the motor.

  The means for detecting the overload of the crushing device is achieved by detecting the driving current of the motor.

The means for detecting the overload of the device is achieved by a strain gauge. The time for reversing the motor after detecting the overload of the crushing device is achieved by changing based on the time interval at which the overload is detected.

  The time for reversing the motor after detecting the overload of the crushing device is achieved by lengthening the time interval at which the overload is detected.

  The time for reversing the motor after detecting the overload of the crushing device is achieved by changing it based on the number of times the overload is detected per certain time.

  The time for reversing the motor after detecting the overload of the crushing device is achieved by increasing the number of times the overload is detected per fixed time.

  By adopting the above-described configuration, the rotation of the screw shaft driven by the motor of the crushing device rotates the frozen biological tissue piece, the tissue containing the frozen smooth muscle, or the hard tissue such as bone. By crushing with a blade, a large amount of sample can be crushed in a short time, and heat during crushing can be minimized. As a result, it is possible to provide a crushing apparatus that is easy to handle, can be processed quickly and safely, and has high crushing efficiency and a high recovery rate of useful substances.

The crushing apparatus of the present invention will be described by taking a cell crushing apparatus as an example with reference to the drawings.
An embodiment of the present invention will be described with reference to FIGS. FIG. 1 is a side view illustrating a structure and a component configuration showing an embodiment of a cell disrupting apparatus. FIG. 2 is a perspective view showing an embodiment of the cell crushing apparatus.

  In FIG. 1, a motor 2 for driving the screw shaft 4 is built in a housing 22 of the cell crushing apparatus 1, and a crushing unit 3 is detachably attached to the housing 22 to constitute the crushing unit 3. A screw shaft 4 is rotatably disposed inside the storage cylinder 7, and a rotary blade 5 disposed to rotate together with the screw shaft 4 is held on the discharge port 21 side of the storage cylinder 7. Yes.

  Furthermore, the fixed blade 6 having the perforated hole 6a provided so as to face the rotary blade 5 is attached so that it cannot be rotated by the fixed blade holder 20 and a rotation preventing member (not shown). Further, the storage cylinder 7 is provided with a supply port 8 for continuously introducing into the crushing section 3 a living tissue piece frozen from a tissue containing smooth muscle such as an animal heart or a hard tissue such as a bone. ing.

  In this embodiment, the screw shaft 4 is installed so as to be detachably fitted to the rotating shaft of the motor 2 via the overload detector 9. 3 is a cross-sectional view taken along the line AA in FIG. The screw shaft 4 has a structure in which the shaft diameter gradually increases toward the discharge port 21, and a plurality of grooves 12 are formed in the direction of the rotation axis of the screw shaft 4 in the housing cylinder 7 that holds the screw shaft 4. .

In the above configuration, when a switch (not shown) of the cell crushing apparatus 1 is turned on, the motor 2 is driven by the output from the motor control unit 10 and the screw shaft 4 and the rotary blade 5 rotate.
In FIG. 4, the frozen biological tissue piece 11 put into the cell crushing device 1 enters a gap formed by the screw shaft 4 and the storage cylinder 7, and the frozen biological tissue piece 11 is screw blades when the screw shaft 4 rotates. It is pushed by 4 a and sent to the discharge port 21 along the plurality of grooves 12. The gap between the screw shaft 4 and the storage cylinder 7 is gradually reduced, and the biological tissue piece 11 frozen with liquid nitrogen or the like receives a compressive force, and is crushed through the processes from a to d in FIG. Extruded in the direction.

  The plurality of grooves 12 in the direction of the rotation axis formed inside the storage cylinder 7 serve to suppress the movement of the frozen biological tissue piece 11 in the circumferential direction along with the rotation of the screw shaft 4, and the frozen and crushed biological tissue. It has a function as a guide groove for feeding the piece 11 to the discharge side.

  The crushed frozen biological tissue piece 11 extruded in the direction of the discharge port 21 is further crushed by the porous hole 6a of the rotary blade 5 and the fixed blade 6, and the biological tissue is frozen or in a sherbet-like state. Then, it is discharged out of the crushing part 3 from the discharge port 21.

  In FIG. 5, when a biological tissue piece 11 containing a very hard tissue (for example, bone) is put into the apparatus and can withstand the compressive force created by the screw shaft 4 and the storage cylinder 7, an excessive load is applied to the motor 2. And the signal from the overload detector 9 is output. Upon receiving a signal from the overload detector 9, the motor control unit 10 immediately stops the rotation of the motor 2, and the screw shaft 4 stops at the position shown in FIG. 5a.

  Thereafter, the motor rotates in reverse to rotate the screw shaft 4 as shown by the arrow in FIG. 5b. By rotating in reverse, the gap between the screw shaft 4 and the storage cylinder 7 becomes large, and the living tissue piece 11 including the hard tissue changes its position and orientation.

  Biological tissues such as animals and plants are highly directional dependent on mechanical strength (for example, tensile strength and compressive strength), and can be crushed with a small force by changing the direction in which the force is applied. If the direction of the biological tissue piece 11 is changed and then the motor 2 is rotated forward again, the biological tissue piece 11 containing a hard tissue can be finely crushed as shown in FIGS. 5c to 5e.

  FIG. 6 shows the timing of reverse rotation of the motor after overload detection, and shows a case where a method of detecting the rotation speed of the motor is used as the overload detector 9. Further, a method of detecting when a strain gauge is used as an overload detector 9 of another method is also described.

  In a normal crushed state body, the motor is operated at a substantially constant rotational speed. However, as shown in FIG. 5b, when the rotation of the screw shaft 4 is remarkably reduced by the biological tissue piece 11, or when the screw shaft 4 is locked, an overload is applied to the motor 2 that rotationally drives the screw shaft 4, and the rotation of the motor Is suppressed, and the signal of the overload detector 9 is turned ON when the rotation speed becomes a predetermined value or less. Upon receiving this signal, the motor control unit 10 stops the rotation of the motor 2 and then performs reverse rotation at a predetermined rotation speed and time. When the reverse rotation is completed, the motor is operated again in the normal rotation. Further, the reverse rotation may be a rotation speed (for example, 1 to 2 rotations).

  The means for detecting the overload may be a method for detecting the drive current of the motor. When an overload is applied, the rotation of the motor is suppressed and the drive current increases. By detecting this and turning on the signal output of the overload detector 9, the same effect can be obtained.

  Furthermore, the output of the strain gauge may be used as a means for detecting overload. For example, the strain gauge is arranged on the motor shaft, and the rotation of the screw shaft 4 is significantly reduced by the biological tissue piece 11 as shown in FIG. 5b. In this case, or in a locked state, a torsion torque is generated between the screw shaft 4 and the motor shaft, and the output from the strain gauge increases. A similar effect can be obtained by turning on the signal output when the output of the strain gauge exceeds a predetermined value (threshold value) as shown in FIG.

  In addition, the position where the strain gauge is disposed need not be the motor shaft. If a torsional torque is generated between the screw shaft 4 and the motor shaft, the motor 2 itself tries to rotate around the motor rotation shaft. For example, it may be a leg portion that supports the motor, or may be a support shaft disposed on the motor rotation shaft between the housing 22 and the motor 2. .

FIG. 7 shows an embodiment in which the motor 2 is reversely rotated several times.
After operating for t1 hours in a steady state, the rotational speed of the motor is reduced to a predetermined rotational speed or less by the biological tissue piece 11 that has not been crushed, and the first ON signal is output from the overload detector 9. . Upon receiving this ON signal, the motor control unit 10 stops the rotation of the motor and then reversely rotates the motor for a predetermined rotation speed and time (T1 time). When the reverse rotation is completed, the motor 2 operates again in the normal rotation.

  After that, when the second ON signal is output again from the overload detector 9, the motor control unit 10 receives this signal and stops the motor, and the first ON signal and the second ON signal. The interval between the signals is t2 hours, and t1 and t2 are compared. As a result of the comparison, when t2 is shorter than t1, there is little change in the direction of the biological tissue piece 11 due to the reverse rotation of the motor 2, so it is considered that a compressive force is applied in the same direction, and crushing processing is performed. This is a problem that does not progress at all.

  Therefore, the motor control unit 10 controls the reverse rotation time of the motor so as to reversely operate the motor for T2 time longer than T1 time, and can change the direction of the biological tissue piece 11 greatly, The possibility that the tissue piece 11 can be crushed is improved.

Further, when the third ON signal is output from the overload detector 9, the motor control unit 10 stops the motor 2 and sets the interval between the second ON signal and the third ON signal. When t3 is set to be shorter than t2, the reverse rotation time of the motor is controlled to reversely run for T3 time longer than T2 time.
By performing such control, the direction of the biological tissue piece 11 can be changed further greatly, and the progress of the crushing process can be prevented from stopping.

  FIG. 8 shows the relationship between the overload detection time interval and the reverse rotation time. By increasing the reverse rotation time exponentially as the overload detection time interval becomes shorter, it is possible to prevent the crushing process from stopping. For example, the motor reverse rotation time when the overload detection time interval is t1 is T1, and the motor reverse rotation time when the overload detection time interval is t3 is T3. Alternatively, as shown in FIG. 9, several types of reverse rotation time may be set in advance, and the reverse rotation time may be increased stepwise as the overload detection time interval becomes shorter.

  Further, the same effect can be obtained by changing the time for reversing the motor according to the number of times of overload detection within a certain time instead of the overload detection time interval. In FIG. 10, when the number of overload detections within a certain time is counted n times, the direction of the biological tissue piece 11 is little or not changed due to the reverse rotation of the motor 2, so the compression force is continuously applied in the same direction. It can be considered that the crushing process does not proceed at all.

  In such a case, the direction of the biological tissue piece 11 can be changed largely from the previous time by performing the reverse rotation of the motor 2 for a longer time than the previous reverse rotation time, and the biological tissue piece 11 is crushed by the forward rotation again. The possibility of being improved. In FIG. 11, the progress of the crushing process can be prevented by increasing the reverse rotation time as the number of detections increases.

  Furthermore, when the overload detection interval becomes shorter than a predetermined time, or when the number of overload detection reaches a predetermined number of times within a certain time, the motor control unit 10 It is determined that crushing is impossible, the motor 2 driving the screw shaft 4 is stopped, and the motor 2 cannot crush the biological tissue piece and is stopped due to overload. Display with the indicator.

By adopting such a configuration, the frozen biological tissue piece is reliably crushed in a very short time, so that the influence of heat during crushing is minimized, and the biological tissue piece is smaller than that of a rotary homogenizer. Even if it is not damaged by the rotating blade that rotates at high speed, and the recovery rate is reduced due to alteration of heat-sensitive substances such as protein, DNA, RNA due to frictional heat, or extraction of proteins with large molecular weight, Damage to collected materials such as proteins has been reduced, leading to a dramatic increase in the rate of extraction of biological tissues. For example, when microsomes were extracted from human liver, the ability of these microsomes to metabolize drugs was improved by several percent compared to those extracted by conventional methods. Furthermore, in the conventional cell crushing apparatus, it was not possible to continuously put and crush biological tissue pieces, but in the present invention, it is also possible to continuously feed samples such as biological tissue pieces from the supply port 8, Compared to conventional cell disrupters, a significant increase in throughput was achieved, enabling industrial use by pharmaceutical companies and the like.
Furthermore, by controlling the motor of the present invention, the motor can be reduced in size, so that the crushing device can be reduced in size and carried.

It is a side view explaining the structure and component structure of a cell crushing apparatus which shows one Example of this invention. It is a perspective view of the cell crushing apparatus which shows one Example of this invention. It is AA sectional drawing of the cell crushing apparatus by one Example of this invention. It is a figure which shows a mode that the frozen biological tissue piece in the AA cross section is crushed. It is a figure which shows a mode that the frozen biological tissue piece in the AA cross section is crushed. It is a figure which shows the timing which reversely rotates a motor by overload. It is a figure which shows the change of the time which reversely rotates a motor by overload. It is a figure which shows the relationship between an overload detection interval and motor reverse rotation time. It is a figure which shows the relationship between an overload detection interval and motor reverse rotation time. It is a figure which shows the change of the time which reversely rotates a motor by the frequency | count of overload detection. It is a figure which shows the relationship between the frequency | count of overload detection, and motor reverse rotation time. It is sectional drawing of the conventional cell crushing apparatus.

Explanation of symbols

DESCRIPTION OF SYMBOLS 1 Cell crushing apparatus 1, 2 Motor, 3 Crushing part, 4 Screw shaft, 4a screw blade, 5 Rotary blade, 6 Fixed blade, 6a Porous hole, 7 Storage cylinder, 8 Supply port, 9 Overload detector, 10 Motor control 11, frozen biological tissue piece, 12 groove, 20 fixing holder, 21 discharge port, 22 housing

Claims (8)

A housing, a motor disposed in the housing, a storage cylinder having a supply port and a discharge port, and a screw shaft driven by the motor are rotatably supported in the storage cylinder, and the discharge of the storage cylinder In the crushing apparatus, a rotating blade that rotates together with the screw shaft is attached to the side, and a fixed blade is attached to the storage cylinder side so as to face the rotating blade. A crushing apparatus comprising: a means for reversing the motor by an output signal from the means for detecting the load. The crushing apparatus according to claim 1, wherein the means for detecting an overload of the cell crushing apparatus detects a rotational speed of a motor. The crushing apparatus according to claim 1, wherein the means for detecting an overload of the cell crushing apparatus detects a driving current of a motor. The crushing apparatus according to claim 1, wherein the means for detecting an overload of the cell crushing apparatus is a strain gauge. The crushing apparatus according to claim 1, wherein the time for reversing the motor after detecting the overload of the cell crushing apparatus is changed based on the time interval at which the overload is detected. 6. The crushing device according to claim 5, wherein the time for reversing the motor after detecting the overload of the cell crushing device is increased as the time interval at which the overload is detected becomes shorter. The crushing apparatus according to claim 1, wherein the time for reversing the motor after detecting the overload of the cell crushing apparatus is changed based on the number of times the overload is detected per fixed time. 8. The crushing device according to claim 7, wherein the time for reversing the motor after detecting the overload of the cell crushing device is lengthened as the number of overload detections per fixed time increases.

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