JP4279545B2 - Crushing and centrifugation methods - Google Patents

Description

[0001]
BACKGROUND OF THE INVENTION
The present invention relates to a crushing and centrifuging method for crushing and centrifuging and analyzing plant tissues and seeds, animal tissues, microorganisms, molds, plastic materials, mineral materials, and the like for chemical analysis and fractional separation. It is.
[0002]
[Prior art]
In order to chemically analyze and fractionate the above various materials, the materials must first be uniformly crushed. Generally, a mortar and a pestle are used, but it takes time and labor to obtain a uniform crushed state, and is not efficient. Therefore, a crushing device is used to efficiently perform the crushing work. By causing the crushing container to perform the revolving motion and the rotation motion simultaneously, the crushing medium consisting of the material to be crushed and the microbeads stored in the crushing vessel is three-dimensional. There is known a “planet type mill” that is crushed by crushing by crushing by compression and rotation by collision of the crushing medium against the object to be crushed.
[0003]
In addition, as a cell crushing device for screening of substance-producing bacteria such as yeast and bacteria, production cells, etc., the present applicant accommodates microbeads made of glass or ceramics together with an object to be crushed in a crushing container. Has been proposed in which a high-speed rotation is performed in the shape of an 8-shaped vibration, and efficient collisions between microbeads and cells are repeated in a short time (see Patent Document 1).
[0004]
In addition, by improving the cell disruption apparatus, a single disruption medium and an object to be crushed are accommodated in a disruption container, and an 8-shaped vibration is applied so that the disruption medium is a pestle and the disruption container is a mortar. An efficient crushing device that works like this has been proposed (see Patent Document 2).
[0005]
[Patent Document 1]
Japanese Examined Patent Publication No. 6-36732 (pages 2 and 3, FIG. 1)
[0006]
[Patent Document 2]
JP 2001-178444 A (2nd to 4th pages, FIG. 2)
[0007]
[Problems to be solved by the invention]
When crushing the material to be crushed by the crushing device and performing analysis such as DNA extraction, the buffer solution is injected into the crushing vessel after crushing, and the material to be crushed is suspended in the buffer solution. Need to be centrifuged. Conventionally, after crushing is completed by a crushing device, it is necessary to individually take out a plurality of crushing containers from the crushing device and set them in a centrifuge, and there is a problem that becomes troublesome work when the number of crushing containers is large. .
[0008]
The purpose of the present invention is to provide a crushing and centrifuging method that makes it possible to transfer a container holder that holds a number of crushing containers containing objects to be crushed to a centrifuge after crushing processing by a crushing device. It is to provide.
[0009]
[Means for Solving the Problems]
In order to achieve the above object, according to the present invention, a rotary shaft that is rotationally driven by a drive source is provided with an inclined shaft portion whose axis is inclined with respect to the axis, and an annular body that is rotatable relative to the inclined shaft portion. The crushing medium is a single object that is removably moved in the axial direction in an elongated crushing container using a crushing device provided with elastic restraining means that elastically restrains the rotation of the annular body. And each of the plurality of objects containing the object to be crushed is held in a plurality of container receiving holes provided on the circumference around the center line of the container holder, and then the container holder is attached to the annular body. By attaching the center line of the container holder and the axis of the inclined shaft portion so as not to rotate relative to each other, rotating the rotating shaft, and reciprocating the crushing medium in the crushing container. A crushing step of crushing the material to be crushed; After completion of the crushing process, the crushing medium is taken out from each crushing container with each crushing container held in the container holder, and a buffer solution injecting process for injecting a buffer solution into each crushing container, and a rotational drive by a drive source The container holder holding each crushing container is centerlined using a centrifuge provided with a rotor body that is integrally rotated with the driven shaft and accommodates the container holder so as not to be relatively rotatable. And the drive shaft is accommodated and mounted in the rotor body so that the axis of the drive shaft coincides, and the drive shaft is rotated to centrifuge the object to be crushed in the crushing container after the buffer solution injection step As the container holder, the axis of each of the plurality of container receiving holes intersects with the center line of the container holder at an acute angle above the upper surface of the container holder, and each crushing container is inclined obliquely downward and outward. Using the one that is held in the state, using the one that can be attached to and detached from the annular body by moving the container holder in the axial direction of the inclined shaft portion as the crushing device, A container holder that can be attached to and detached from the rotor body by moving the container holder in the axial direction of the drive shaft is used. According to the present invention, the container holder that holds the plurality of crushing containers is configured to be detachable from the crushing apparatus and the centrifuge, so that when the container holder is taken out after crushing processing by the crushing apparatus and attached to the centrifuge In addition, it is possible to quickly carry out the centrifugation process on the crushed object.
[0010]
[0011]
[0012]
DETAILED DESCRIPTION OF THE INVENTION
Hereinafter, embodiments of the present invention will be described with reference to the accompanying drawings for understanding of the present invention. The following embodiment is an example embodying the present invention, and does not limit the technical scope of the present invention.
[0013]
FIG. 1 shows the configuration of the crushing apparatus according to the first embodiment. A rotating shaft 8 that is rotationally driven by a motor (not shown) is rotatably supported by a pair of bearings 10 of a bearing portion 7. The upper portion extends above the bearing portion 7. An inclined shaft body 11 is fitted to the upper portion of the rotating shaft 8 with the shaft center inclined at an angle θ with respect to the shaft center, and is screwed to the upper end portion of the rotating shaft 8 via the inclined ring 12. The nut 13 is pressed and fixed to the rotary shaft 8.
[0014]
An annular body 15 is attached to the outer periphery of the inclined shaft body 11 via a bearing 14 so as to be relatively rotatable. A magnet 16 is attached to the annular body 15, and a counter electrode magnet 18 is attached to a support column 17 fixed so as to face the magnet 16. When the rotating shaft 8 and the inclined shaft body 11 are rotated by the attractive force between the magnets 16 and the counter electrode magnet 18, the rotation of the annular body 15 that follows the rotation shaft 8 and the inclined shaft body 11 is prevented. Thus, the annular body 15 is configured to perform a swinging motion. The magnet 16 is provided at a plurality of locations on the circumference of the annular holding body 20, and the counter electrode magnet 18 is provided so as to face each magnet 16, thereby preventing the rotation of the eccentric annular body 15 more reliably. Can be done.
[0015]
A ring-shaped container holder 22 is attached to the holding body 20 fixed to the annular body 15. As shown in FIG. 2, the container holder 22 is formed with a plurality of container accommodation holes 50 for accommodating the crushing containers 30 at a predetermined inclination angle on the circumference, and below the cylindrical portion 20a of the holding body 20 at a central position. A fitting insertion hole 24 in which a spline hole corresponding to the spline (not shown) is formed is provided. Although not shown in the drawing, the spline corresponding to the insertion hole 24 of the container holder 22 is formed on the outer periphery of the cylindrical portion 20a, and the male screw for screwing the fixing cap 25 is formed on the upper side. .
[0016]
The container holder 22 accommodates the crushing container 30 closed by the lid body 31 by putting the object to be crushed and the crushing medium into the plurality of container receiving holes 50, and as shown in FIG. Each crushing device accommodated in the container holder 22 by being inserted into the crushing device by being inserted into the portion 20a, fitting the pressing plate 23 shown in FIG. 3 into the cylindrical portion 20a, and screwing the fixing cap 25 into the cylindrical portion 20a. The container 30 is fixed to the container holder 22 by the lid body 31 being pressed by a tab 23 a provided on the pressing plate 23, and the container holder 22 is also fixed to the annular body 15 by a fixing cap 25.
[0017]
When the crushing apparatus to which the container holder 22 is attached is operated, an 8-shaped shake motion is applied to each crushing container 30 and the object to be crushed is crushed by the crushing medium. That is, as shown in FIG. 4A, when the state where the annular body 15 is inclined to the right side is a reference position and the behavior of the point a position on the outer periphery of the annular body 15 at that time is seen, When the rotary shaft 8 rotates 90 ° in the direction of the arrow, the annular body 15 shifts to a state inclined in the front and back direction of the paper surface as indicated by the phantom line, while the position corresponding to the point a passes through the path b and becomes c Move to a point. Next, when the rotating shaft 8 is further rotated by 90 °, the annular body 15 shifts to the left side as shown by a solid line in FIG. 4B, and the position corresponding to the point a is a path from the point c. Return to the original point a through d. When the rotating shaft 8 further rotates 90 °, the annular body 15 shifts to a state inclined in the opposite direction to the front and back of the paper surface as shown by the phantom line, and the position corresponding to the point a passes through the path e and becomes the point f. When the rotary shaft 8 is further rotated 90 ° to the original rotational position, the position corresponding to the point a returns from the point f through the path g to the original point a. Accordingly, the rotation of the rotating shaft 8 causes the arbitrary position of the outer peripheral portion of the annular body 15 to repeat a figure- 8 swinging motion, and the crushing container 30 accommodated in the container holder 22 fixed to the annular body 15. In addition, an 8-shaped swing motion is applied to the crushing medium accommodated in the crushing container 30 to crush the object to be crushed.
[0018]
As shown in FIG. 5, the crushing container 30 is formed of an elongated cylindrical container. A screw 30a is formed on the outer periphery of the opening, a head hemisphere 35 is formed on the bottom, and a lid 31 is formed on the opening. It is comprised so that it can be sealed by screwing together. An annular seal portion 31 a that fits to the inner periphery of the opening of the crushing container 30 is formed on the inner periphery of the lid 31. The crushing container 30 has a capacity of 2 ml to 50 ml depending on the material and amount of the material to be crushed, and the container holder 22 is also formed with a container receiving hole 50 corresponding to the size of the crushing container 30. Used.
[0019]
The crushing medium 32 accommodated in the crushing container 30 together with the material to be crushed is constituted by a single member having a length L larger than the inner diameter D of the crushing container 30 as shown in FIG. A similar rounded spherical protruding end 32 a is formed at one end corresponding to the shape of the bottom of the crushing container 30. The other end portion is formed in the small diameter portion 32b so as not to interfere with or fit into the annular seal portion 31a on the inner periphery of the lid body 32. Further, the outer diameter d of the crushing medium 32 is set to be 2 mm or less with respect to the inner diameter D of the crushing container 30, and about 1 mm or less when the inner diameter d is small. For example, when the capacity of the crushing container 30 is 2 ml, the inner diameter D is set to 8 mm, and the outer diameter d of the crushing medium 32 is set to 7 mm.
[0020]
Further, as shown in FIGS. 6B and 6C, by forming one or a plurality of grooves 34 radially or spirally in the protruding end portion 32 a of the crushing medium 32 as necessary, Since the fiber that is difficult to cut is effectively cut, it can be efficiently crushed.
[0021]
Moreover, the crushing medium 32 is formed corresponding to the shape of the crushing container 30, and can also be configured as shown in FIGS. 7 and 8A to 8C. Since the crushing container 30 shown in FIG. 7 has a conical conical portion 33 formed at the bottom thereof, the crushing medium 32 has a conical protruding end portion 32a at one end thereof as shown in FIG. Is formed. Further, as shown in FIGS. 8B and 8C, a groove 34 can be formed in the protruding end portion 32a.
[0022]
[0023]
When the material to be crushed is used for analysis after the material to be crushed using the crushing device having the above-described configuration, it is necessary to inject a buffer into the crushing container 30 after crushing and perform centrifugation. . Therefore, when the crushing by the crushing apparatus is completed, the container cap 22 can be taken out by loosening and removing the fixing cap 25 and removing the pressing plate 23. Therefore, when the container holder 22 is transferred to the centrifuge, it is centrifuged as it is. Can be transferred to work.
[0024]
As shown in FIGS. 9 and 10, the centrifuge corresponding to the container holder 22 has a rotor body in which a drive shaft 53 that is rotationally driven by a motor (not shown) is provided with a boss 54 having a spline 55 formed at the center position. 57 is fixed by a bolt 56.
[0025]
Each crushing container 30 that has been crushed by the crushing device opens the lid 31, takes out the crushing medium 32, and injects a buffer solution into the crushing container 30. In addition, the method of taking out the crushing medium 32 from the crushing container 30 can apply the method of taking out the crushing medium 32 taken out simultaneously with the removal of the lid 31, and has been proposed as Japanese Patent Application No. 2002-208108 by the applicant of the present application. .
[0026]
As shown in FIG. 10, the container holder 22 containing the crushing container 30 infused with the buffer solution is attached to the centrifuge as shown in FIG. 10 by inserting the insertion hole 24 provided at the center position into the boss 54. be able to. In this centrifuge, three container holders 22 can be mounted, but one container holder 22 can also be mounted. When the centrifuge is operated with the container holder 22 attached, the container holder 22 also rotates with the rotation of the rotor body 57, and the crushed and crushed buffered material is separated into a supernatant and a precipitate by centrifugation. To separate. The material to be crushed can be analyzed by means such as measuring the absorbance of the centrifuged supernatant.
[0027]
The holding angle of the crushing container 30 accommodated in the container holder 22 has a suitable inclination angle according to the capacity of the crushing container 30, and the inclination angle is generally set smaller as the capacity increases.
[0028]
Next, the crushing apparatus according to the second embodiment will be described. The crushing apparatus according to the second embodiment is different in the mounting structure of the container holder 60, and the same reference numerals are given to the components common to the configuration of the first embodiment, and the description thereof is omitted.
[0029]
In FIG. 11, the crushing apparatus according to the second embodiment includes a container holder 60 formed in a donut shape in an annular body 15, and a pressing plate 61 is disposed on the annular body 15. By screwing the screw portion into the annular body 15, the container holder 60 is fixed to the annular body 15, and the crushing container 30 accommodated in the container holder 60 is held in the container accommodation hole 65.
[0030]
As shown in FIG. 12, the container holder 60 is formed with a plurality of container accommodation holes 65 on the circumference for accommodating a plurality of crushing containers 30 at a predetermined inclination angle, similar to the container holder 22 in the first embodiment. It is formed in a donut shape having an opening at the center. Further, as shown in FIG. 13, the pressing plate 61 is formed with a tab 66 that presses and holds the lid 31 of each crushing container 30, and the base of the tab 66 presses and fixes the container holder 60 to the annular body 15. The container holder 60 is pressed and fixed by screwing the threaded portion of the fixing knob 62 into the annular body 15 from the fixing hole 67.
[0031]
As shown in FIG. 14, the centrifuge corresponding to the container holder 60 configured as described above includes a rotor body 70 that houses the container holder 60. The container holder 60 that has been attached to the crushing apparatus as described above and has been crushed can be removed from the crushing apparatus by loosening the fixing knob 62 and stored in the rotor body 70 as shown in the figure.
[0032]
The centrifuge is configured such that the rotor main body 70 is rotationally driven by a motor 71, and the rotation of the rotor main body 70 also rotates the container holder 60 accommodated therein, so that centrifugal force is applied to each crushing container 30. Therefore, if the crushed material to be crushed is suspended in a buffer solution, it can be separated into a supernatant and a precipitate by centrifugation.
[0033]
The holding angle of the crushing container 30 by the container holder 60 has a suitable inclination angle according to the capacity of the crushing container 30 when performing centrifugation, and the inclination angle is generally set smaller as the capacity increases.
[0034]
【The invention's effect】
As described above, according to the present invention, a plurality of crushing containers containing a material to be crushed and a crushing medium can be accommodated in a container holder, and a lot of materials to be crushed can be crushed at the same time. After completion, the container holder can be transferred to the centrifuge, so that the crushing containers can be individually transferred from the crushing device to the centrifuge in operations such as analysis where centrifuge processing is indispensable after crushing the material to be crushed. The troublesome work of transferring can be eliminated.
[Brief description of the drawings]
FIG. 1 is a cross-sectional view showing the main configuration of a crushing apparatus according to a first embodiment.
FIG. 2 is a plan view showing a configuration of a container holder according to the first embodiment.
FIG. 3 is a plan view showing a configuration of a pressing plate according to the first embodiment.
FIG. 4 is an explanatory diagram of an 8-shaped reciprocating vibration.
FIG. 5 is a cross-sectional view showing a configuration of a crushing container.
FIG. 6 is a front view showing various examples of crushing media to be combined with the crushing container.
FIG. 7 is a cross-sectional view showing a configuration of a crushing container.
FIG. 8 is a front view showing various examples of crushing media to be combined with the crushing container.
FIG. 9 is an exploded perspective view showing the main configuration of the centrifuge according to the first embodiment.
FIG. 10 is a cross-sectional view of the centrifuge.
FIG. 11 is a cross-sectional view showing the main configuration of a crushing apparatus according to a second embodiment.
FIG. 12 is a plan view showing a configuration of a container holder according to a second embodiment.
FIG. 13 is a plan view showing a configuration of a pressing plate according to a second embodiment.
FIG. 14 is a cross-sectional view showing a configuration of a centrifuge according to a second embodiment.
[Explanation of symbols]
DESCRIPTION OF SYMBOLS 1, 2 Crushing device 8 Rotating shaft 11 Inclined shaft part 15 Annular body 16 Magnet 18 Counter electrode magnet 20 Holding body 22, 60 Container holder 23, 61 Press plate 25 Fixed cap 30 Crushing container 50, 65 Container accommodation hole 62 Fixed knob

Claims (1)

  A rotating shaft that is rotationally driven by a drive source is provided with an inclined shaft portion whose axis is inclined with respect to the axial center, and an annular body is externally fitted to the inclined shaft portion so as to be rotatable relative to the rotating shaft. Using a crushing device provided with elastic restraining means for elastically restraining the material, a single crushing medium and a material to be crushed accommodated in a slender crushing container that can reciprocate in its axial direction Each of the books is held in each of a plurality of container receiving holes provided on a circumference around the center line of the container holder, and then the container holder is attached to the annular body with the center line of the container holder and the inclined axis. A crushing step of crushing the material to be crushed by attaching the shafts of the parts so as not to rotate relative to each other, rotating the rotary shaft, and reciprocating the crushing medium in the crushing container;
  After completion of the crushing step, in a state where each crushing container is held in a container holder, the crushing medium is taken out from each crushing vessel, and a buffer solution injection step for injecting a buffer solution into each crushing vessel,
  The container holder that holds each crushing container using a centrifuge provided with a rotor body that rotates integrally with a drive shaft that is rotationally driven by a drive source and that is provided so as to accommodate the container holder in a relatively non-rotatable manner. The center line and the axis of the drive shaft coincide with each other so that the rotor body is housed and mounted, and the drive shaft is rotated, whereby the object to be crushed in the crushing container after the buffer solution injection step Centrifuge the object,
  As the container holder, the axis of each of the plurality of container receiving holes intersects with the center line of the container holder at an acute angle above the upper surface of the container holder, and holds the crushing containers obliquely downward and outward. Use
  As the crushing device, the container holder can be attached to and detached from the annular body by moving in the axial direction of the inclined shaft portion,
  As the centrifugal separator, a device that can be attached to and detached from the rotor body by moving the container holder in the axial direction of the drive shaft was used.
A crushing and centrifuging method characterized in that.

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