JP4373391B2 - Sample crushing tool - Google Patents

Description

  The present invention relates to a sample crushing tool used for crushing a sample of plant tissue, seeds, animal tissue, plastic material, mineral material or the like for chemical analysis and fractional separation.

  In order to chemically analyze and fractionate various samples as described above, the sample must first be uniformly pulverized. Conventionally, crushing using a mortar and pestle is known, but since it is not suitable for efficiently crushing many samples, it is not suitable for crushing containers containing samples and crushing media. A crushing apparatus that crushes many at the same time is used.

  In order to crush a sample using the crushing apparatus, a crushing medium is stored together with the sample in a crushing container such as a centrifuge tube that is generally used for centrifuging the sample, and the crushing container is used as a crushing apparatus. Wear and apply vibration to the crushing container. As the crushing medium, fine beads such as glass, ceramic, and metal are known (see Patent Document 1). When vibration is applied to the crushing container by the crushing device, the crushing medium collides with the sample or is pressed against the wall surface of the crushing container and is crushed. Therefore, the sample is crushed by applying vibration for the required time to the crushing container.

A frozen sample crushing cell is known as an instrument for crushing a sample (see Patent Document 2). This is done by freezing the sample in order to make it easy to crush the sample, putting it into a cooled bottomed cylindrical container, and installing a lid with a protruding part having a shape corresponding to the inner shape of the container. Insert the sample into the container, put it on the container, apply a vibration press to the lid, and grind the frozen sample.
Japanese Examined Patent Publication No. 6-036732 (pages 2 and 3, FIG. 1) JP-A-8-219957 (pages 1 and 2, FIG. 1)

  However, in the crushing method using microbeads as a crushing medium, when the sample becomes a large plant cell, animal cell, mineral material, etc., there is a problem that the sample is difficult to move and the crushing medium is not crushed because the mass of the crushing medium is small. there were.

  In addition, in order to use the crushed sample for processing such as centrifugation and analysis, the frozen sample crushing cell needs to transfer the crushed sample from the container to another container necessary for the processing. If the sample contains bacteria or the like, it may be dissipated. In addition, there is a problem that operations such as washing and sterilization are required for the next crushing.

  The present invention has been made in view of the above-described conventional problems, and an object thereof is to provide a sample crushing tool capable of efficiently crushing large plant tissues, animal tissues, mineral materials, and the like.

The first invention of the present application for achieving the above object is a sample crushing tool for storing a crushing medium together with a sample to be crushed in a crushing container, and for crushing the sample by vibrating the crushing container. A container body having a bottom cylindrical shape with a bottom surface recessed in a substantially hemispherical shape or a semi-elliptical spherical shape, and a lid having a ceiling surface recessed in a substantially hemispherical shape or a semi-elliptical spherical shape so that the opening can be opened and closed. A crushing container provided with a body and a cylindrical body that is accommodated in the crushing container and the main body portion holds a posture substantially along the axis in the container main body and moves relatively in the axial direction. One end portion is substantially hemispherical or semi-elliptical spherical so as to correspond to the shape of the bottom surface of the container main body, and is projected so as to be continuous with the main body portion, and the other end portion is a ceiling of the lid body. Is it approximately hemispherical or semi-elliptical to correspond to the surface shape? Wherein is protrusively manner continuous to the body portion, the length is longer than the inner diameter of the crushing chamber, the outer diameter of 1mm to 2mm smaller grinding media than the inner diameter of the crushed container when the volume of the crushing chamber is 2ml to 50ml Is provided.

  When the crushing medium is accommodated together with the sample in the crushing container, and the reciprocating vibration is applied to the crushing medium in a direction substantially along the axis thereof, preferably the reciprocating vibration in the shape of 8 is applied, Since the operation of colliding with the bottom surface is repeated, the crushing container acts like a mortar and the crushing medium acts like a pestle so that the sample is effectively crushed. Even if the sample is a large plant tissue, animal tissue, plastic material or mineral material, it can be efficiently crushed.

The second invention of the present application is a sample crushing tool for storing a crushing medium together with a sample to be crushed in a crushing container, and for crushing the sample by vibrating the crushing container. The container body recessed in a substantially hemispherical shape, a semi-elliptical spherical shape, or a truncated cone shape, and its opening are sealed so that it can be opened and closed, and the ceiling surface is recessed in a substantially hemispherical shape, a semi-elliptical spherical shape or a truncated cone shape. A crushing container provided with a lid, and a circular body so that the main body is accommodated in the crushing container and the main body portion is relatively moved in the axial direction while maintaining a posture substantially along the axis in the container main body. It is formed in a columnar shape, and has one end portion that is substantially hemispherical, semi-elliptical spherical, or conical so as to correspond to the shape of the bottom surface of the container main body, and is provided so as to be continuous with the main body portion. Almost hemispherical, semi-elliptical so that the end corresponds to the ceiling surface shape of the lid And a Jo or frusto conical so as to continuously to said body portion being projectingly, the total length is longer than the inner diameter of the crushing chamber, the outer diameter of the crushed container when the volume of the crushing chamber is 2ml to 50ml And a crushing medium smaller than the inner diameter by 1 mm to 2 mm . According to this, since the sample can be crushed on both the bottom surface and the ceiling surface of the crushing container, more effective crushing can be performed.

  In the above configuration, when the difference between the inner diameter of the crushing container and the outer diameter of the crushing medium is 2 mm or less, even when the sample is supple, the crushing medium can be acted reliably and can be efficiently crushed.

  Further, by forming one or a plurality of grooves on the surface of at least one end of the crushing medium, the fiber is effective in the grooves even in the case of animal tissues or plant tissues that contain fibers that are difficult to cut. Disconnected.

  In addition, if the sample is frozen in a liquid nitrogen bath and freeze-dried, even if the sample contains fibers that are supple and difficult to cut, the sample will be easily cut by freezing, and the sample will be crushed. Is made efficient.

  In addition, if the crushing medium is made of titanium whose surface is polished, the sample can be crushed even in animal tissues such as bone, and tissue such as DNA and RNA remains on the surface of the crushing medium, This eliminates the possibility of cross contamination occurring during crushing.

  As described above, according to the sample crushing tool according to the present invention, the crushing medium is housed together with the sample in the crushing container, and the crushing medium is rotated relative to the bottom surface of the crushing container by reciprocating vibration. Or since it collides repeatedly with a bottom face and a ceiling surface, a crushing container acts like a mortar and a crushing medium acts like a pestle, and a sample is crushed effectively. Even if the sample is a large plant tissue, animal tissue, plastic material, mineral material, etc., it can be efficiently crushed.

Embodiments and reference examples of the present invention will be described below with reference to the accompanying drawings. The present embodiment is an example embodying the present invention, and does not limit the technical scope of the present invention.

1 and 2 show configurations of the crushing container 30 and the crushing medium 32 according to Reference Example 1. FIG. The crushing container 30 is formed of an elongated cylindrical container, the screw 30a is formed on the outer periphery of the opening, the truncated cone part 33 is formed on the bottom, and the lid 31 can be screwed into the opening to be sealed. It is configured. 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 sample.

The crushing medium 32 accommodated together with the sample in the crushing container 30 is composed of a single member having a length L larger than the inner diameter D of the crushing container 30 as shown in FIG. similar truncated conical projecting end portions 32a corresponding to the bottom shape of the crushing chamber 30 is formed in part. 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 is set to about 1 mm or less when the inner diameter D of the crushing container 30 is small. For example, when the capacity of the crushing container 30 is 2 ml and the inner diameter D is 8 mm, the outer diameter d of the crushing medium 32 is set to 7 mm.

  Further, as shown in FIGS. 2B and 2C, the protruding end 32a of the crushing medium 32 may be formed with one or a plurality of grooves 34 formed radially or spirally as necessary. .

4 and 5 show the crushing container 30 and the crushing medium 32 according to the first embodiment. The crushing container 30 shown in FIG. 4 has a hemispherical or semi-elliptical spherical hemispherical portion 35 formed at the bottom thereof, and accordingly, at one end of the crushing medium 32, as shown in FIG. A protruding end portion 32a is formed. Further, as shown in FIGS. 5B and 5C, a groove 34 can be formed in the protruding end portion 32a.

As a material of the crushing container 30 shown in the reference example 1 and the first embodiment, synthetic resins such as polycarbonate, polypropylene, polyethylene, polystyrene, and polyfluorinated ethylene can be used, and it is not necessary to add a colorant. Therefore, the resin is molded to be transparent or translucent. When the crushing container 30 is transparent or translucent, it is suitable for the case where crushing is performed while checking the state of the sample accommodated in the crushing container 30, particularly the state of progress of crushing.

  For example, the screw mouth type centrifuge tube manufactured by Sarstedt (SARSTED: Germany), the Safelock tube manufactured by Eppendorf (Eppendorf: Germany), the centrifuge tube manufactured by Norgen (NALGEN: USA), and the like.

  Further, the crushing container 30 may be made of a metal such as stainless steel or a polyfluorinated ethylene coating on the inner surface thereof.

  As the material of the crushing medium 32, magnetic stainless steel such as SUS430, stainless steel such as SUS304 hard stainless steel, carbon steel, titanium, tungsten, zirconia, tungsten carbide, ceramic, glass, polyfluoroethylene, etc. are used alone or in combination. What was comprised can be used suitably.

When the sample is crushed using the crushing container 30 and the crushing medium 32 shown in the reference example 1 and the first embodiment, as shown in FIGS. The crushing medium 32 and the sample 74 having a longer length are accommodated, the opening of the crushing container 30 is closed with the lid 31, and vibration is applied to the crushing container 30.

  The vibration applied to the crushing container 30 is preferably applied so that the crushing medium 32 repeats collision with the bottom of the crushing container 30 while relatively rotating while maintaining the posture substantially along the axis of the crushing container 30. As a means for realizing the above, when the crushing container 30 is attached to the crushing apparatus and a figure-shaped vibration is applied, the crushing process can be efficiently performed on a large number of crushing containers 30 simultaneously.

  FIG. 7 shows a configuration of a main part of a crushing apparatus capable of applying an 8-shaped vibration to a large number of crushing containers 30. An inclined shaft body 11 is fixed to a rotary shaft 8 that is rotationally driven by a driving means (not shown), and an annular body 15 is attached to the outer periphery of the inclined shaft body 11 via a pair of bearings 14 so as to be relatively rotatable. Yes. A magnet 16 is attached to an appropriate position at the lower periphery of the annular body 15. Further, a counter electrode magnet 18 supported by a mounting bracket 17 attached to the upper portion of the bearing portion 7 is fixedly disposed so as to face the magnet 16. The attractive force of the magnet 16 and the counter electrode 18 prevents the rotation of the annular body 15 when the rotating shaft 8 and the inclined shaft body 11 rotate, and the annular body 15 swings as the inclined shaft body 11 rotates. Is configured to do.

  An attachment step portion 19 is provided on the upper portion of the annular body 15, and an annular holding body 20 is placed thereon and fastened and fixed by a fixing bolt 21. As shown in a plan view in FIG. 8, a large number of elongated container cases 22 are arranged on the outer periphery of the annular holder 20 in a posture parallel to the axis of the annular body 15, and the crushing container 30 is placed in the container case 22. It is configured to accommodate and support. Further, the lid 31 of the crushing container 30 is engaged with the upper end of the container case 22, and a plurality of pressing plates 23 for pressing the upper surface of the lid 31 to fix the crushing container 30 are provided on the annular holding body 20. It is configured to be installed on the provided mounting boss 24 and fastened and fixed with a fixing screw 25. The height of the mounting boss 24 is set to be substantially the same height as the upper surface height position of the lid 31.

  When the crushing container 30 is accommodated in the container case 22 of the annular holder 20 and fixed by the pressing plate 23 in the crushing apparatus having the above-described configuration, and the rotary shaft 8 is driven to rotate by a driving means (not shown), the crushing container 30 is moved to its axis. A reciprocating vibration is formed in an 8-shape that combines a main reciprocating movement of a relatively long stroke in the core direction and a sub-reciprocating movement of a relatively short stroke in the direction orthogonal thereto, and the crushing medium 32 is accompanied by the axial core of the crushing container 30. The crushing container 30 acts like a mortar, the crushing medium 32 acts like a pestle, and the object to be crushed is a large plant cell or animal. Even tissue, plastic materials and mineral materials are efficiently crushed.

The crushing container 30 and the crushing medium 32 can be configured as shown below in addition to the configurations shown in Reference Example 1 and the first embodiment.

9 and 10 show a crushing container 30 and a crushing medium 32 according to Reference Example 2. FIG. Crushing the container 30 shown in FIG. 9, the truncated conical portion 33 to both the bottom and the lid 31 are formed, the grinding media 32 with it, as shown in FIG. 10 (a), similarly to the opposite ends frustoconical projecting end 32a of are formed. Further, as shown in FIGS. 10B and 10C, one or a plurality of radial or spiral grooves 34 can be formed over the entire length of the protruding end portion 32a and the crushing medium 32 as necessary.

12 and 13 show a crushing container 30 and a crushing medium 32 according to the second embodiment. The crushing container 30 shown in FIG. 12 has a hemispherical portion 35 formed on both the bottom portion and the lid 31, and a similar hemispherical protruding end portion 32 a is formed on both ends of the crushing medium 32. Further, as shown in FIGS. 13B and 13C, one or a plurality of radial or spiral grooves 34 can be formed over the entire length of the protruding end 32a and the crushing medium 32 as necessary.

9, as shown in FIG. 12, both ends of the crushing chamber 30 including the lid 31 and the frusto-conical portion 33 and the hemispherical portion 35 and the semi-elliptical bulb, etc., the end portion outer surface facing the grinding media 32 When the shape is substantially the same, the mortar-pestle action can be obtained more effectively, and more efficient crushing can be achieved.

  In the crushing of the sample by the crushing container 30 and the crushing medium 32 described above, if the difference between the inner diameter of the crushing container 30 and the outer diameter of the crushing medium 32 is 2 to 1 mm or less, the crushing medium can be obtained even when the object to be crushed is supple. 32 can be made to act reliably and can be efficiently crushed.

  In addition, in the case of animal and plant tissues that contain fibers that are pliable and difficult to cut, the crushed object is frozen in a liquid nitrogen bath with the crushed container 30 accommodated in the crushing container 30. However, when freeze-drying, the material to be crushed is easily crushed, so that it can be efficiently crushed. In the present invention, the same effect can be obtained only by cooling with dry ice. Furthermore, by cooling the crushing medium 32 together with the object to be crushed with liquid nitrogen or the like, heat generation during crushing can be prevented, and if the crushing medium 32 is cooled to a lower temperature than the object to be crushed, the heat generation is prevented. The effect is remarkable.

  Further, when the object to be crushed is a similar animal or plant tissue, grooves 34 are provided as shown in FIGS. 2 and 4 (b) and (c), and FIGS. 10 and 13 (b) to (e). When the crushing medium 32 is used, the fiber that is difficult to be cut by the groove 34 is effectively cut without being freeze-vacuum dried as described above, so that the crushing medium 32 can be efficiently crushed. In this case, a significant effect is exhibited by cooling with dry ice as described above.

  In addition, when a material made of titanium whose surface is polished is used as the crushing medium 32, a composition such as DNA or RNA is present on the surface of the crushing medium 32 even when the object to be crushed is a hard animal tissue such as bone. It is possible to eliminate the risk of cross contamination occurring during the subsequent crushing.

  In addition, by using a material obtained by coating the surface of a metal material such as carbon steel with polyfluorinated ethylene as the crushing medium 32, the crushing container 30 has a corrosion resistance when containing a corrosive buffer solution or an extract. It is possible to secure the necessary weight for the action.

Sectional drawing which shows the structure of the crushing container which concerns on the reference example 1. FIG. The side view which shows the structure of the crushing medium which concerns on the reference example 1. FIG. Sectional drawing which shows the state which accommodated the crushing medium same as the above in the crushing container. Sectional drawing which shows the structure of the crushing container which concerns on 1st Embodiment. The side view which shows the structure of the crushing medium which concerns on 1st Embodiment. Sectional drawing which shows the state which accommodated the crushing medium same as the above in the crushing container. The principal part sectional drawing which shows the structure of a crushing apparatus. The 1/2 top view which shows the structure of a cyclic | annular holding body. Sectional drawing which shows the structure of the crushing container which concerns on the reference example 2. FIG. The side view which shows the structure of the crushing medium which concerns on the reference example 2. FIG. Sectional drawing which shows the state which accommodated the crushing medium same as the above in the crushing container. Sectional drawing which shows the structure of the crushing container which concerns on 2nd Embodiment. The side view which shows the structure of the crushing medium which concerns on 2nd Embodiment. Sectional drawing which shows the state which accommodated the crushing medium same as the above in the crushing container.

30 crushing container 31 lid 32 crushing medium 33 truncated cone part 34 groove 35 hemisphere part

Claims (6)

In a sample crushing tool that contains a crushing medium together with a sample to be crushed in a crushing container and is used to crush the sample by vibrating the crushing container, it has an elongated bottomed cylindrical shape, and the bottom surface is substantially hemispherical or semi-elliptical spherical A crushing container provided with a lid body that is opened and closed so that the opening thereof is openable and the ceiling surface is recessed in a substantially hemispherical shape or a semi-elliptical spherical shape, and accommodated in the crushing vessel The main body is formed in a cylindrical shape so as to move in the axial direction while maintaining a posture substantially along the axial center in the container main body, and one end is formed in the bottom shape of the container main body. It is substantially hemispherical or semi-elliptical spherical so as to correspond to the main body, and the other end is substantially hemispherical or semi-elliptical so as to correspond to the shape of the ceiling of the lid. in and out as continuous to the main body portion is projectingly, The length is longer than the inner diameter of the crushing chamber, the outer diameter of the sample breaking tool, characterized in that the volume of the crushing chamber is provided with said crushing chamber 1mm to 2mm smaller grinding media than the inner diameter of the case is 2ml to 50 ml. A sample crushing tool that contains a sample to be crushed together with a sample to be crushed in a crushing vessel, and is used to crush the sample by vibrating the crushing vessel. Or a container body recessed in a mounting cone shape and a lid body that seals the opening thereof so that it can be opened and closed, and the ceiling surface is recessed in a substantially hemispherical shape, a semi-elliptical spherical shape, or a mounting cone shape. The crushing container and the crushing container are accommodated, and the main body is formed in a columnar shape so as to relatively move in the axial direction while maintaining a posture substantially along the axis in the container main body. The end is substantially hemispherical, semi-elliptical, or conical so as to correspond to the shape of the bottom surface of the container main body, and is projected so as to be continuous with the main body. Almost hemispherical, semi-elliptical sphere or head cone to correspond to the ceiling shape In a projecting manner and so as to be continuous with the body portion, the length is longer than the inner diameter of the crushing chamber, the outer diameter of 1mm or less 2mm than the inner diameter of the crushed container when the volume of the crushing chamber is 2ml to 50ml A sample crushing tool comprising a crushing medium. The sample crushing tool according to claim 1 or 2, wherein a difference between the inner diameter of the crushing container and the outer diameter of the main body of the crushing medium is 2 mm or less. The sample crushing tool according to any one of claims 1 to 3, wherein one or a plurality of grooves are formed on an end surface of the crushing medium. The sample crushing tool according to any one of claims 1 to 4, wherein the sample is frozen in a liquid nitrogen bath and freeze-dried. The sample crushing tool according to any one of claims 1 to 5, wherein the crushing medium is made of titanium whose surface is polished.

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