JPH07218398A - Method and instrument for preparation of sample of biological tissue - Google Patents

Abstract

PURPOSE:To pick many thinly cut samples from one block without impairing the characteristics and the functions of a biological tissue at a cell level by fixing a plurality of thin blades having the same shape in parallel at a specified interval, thinly cutting the biological tissue, and pushing out the thin pieces with planar pushing members. CONSTITUTION:For example, a blade 1 is made to be a stainless steel plate having the thickness of about 0.1mm, and a pushing plate 2 is made to be a plastic plate having the thickness of about 0.9mm. Five blades 1 are fixed with a blade fixing member 3. A biological tissue is pushed and cut by using the assembled cutter. A plurality of thin pieces of the biological tissue having the thickness corresponding to the interval of the blades l are cut by one action. The pushing plate 2 is inserted between each interval of the blades 1, and the thin piece: of the tissue is pushed out of the cutter. Then, the thin piece of the biological tissue having the thickness of about 0.9mm can be obtained. It is preferable that hydrophobic liquid is applied on the surface of the cutter.

Description

Detailed Description of the Invention

[0001]

BACKGROUND OF THE INVENTION 1. Field of the Invention The present invention relates to a method and an apparatus for preparing a biological tissue sample, and more particularly to a method and a device capable of preparing a plurality of sliced biological tissue samples while maintaining the characteristics and functions of the tissue at the cellular level. It relates to a device therefor.

[0002]

2. Description of the Related Art One of the methods conventionally used for preparing sliced samples of tissue such as animal brain and liver is to use an instrument as shown in FIG. FIG. 3A is an exploded perspective view of the instrument, and B is a longitudinal sectional view. This device is composed of three plate-like members 201, 202 and 203, and the plate-like member 201 has a cylindrical protrusion 204 in the central portion. The plate member 202 has a hole 20 through which the cylindrical protrusion 204 can pass.
5 are provided. The height of the cylindrical protrusion 204 is slightly lower than the thickness of the plate member 202. Plate member 20
3 is provided with a recess 206 penetrating in the width direction,
A gap is formed between the plate member 202 and the plate member 203. Further, the bottom surface of the recess 206 is provided with a cylindrical recess 207 having substantially the same size as the hole 205. The plate member 203 is fixed to the plate member 202 with screws 208.

To prepare a thin sliced sample of a tissue, the plate-shaped member 203 is fixed to the plate-shaped member 202 and the plate-shaped member 2 is used.
01 is separated from the plate-like member 202, and a tissue sample 209 such as a liver is placed on the top of the cylindrical protrusion 204,
The plate-like member 201 and the plate-like member 202 are superposed so that the sheet is inserted into the hole 205. The tissue sample 209 is a cylindrical protrusion 20.
4 is pushed toward the recess 207. While pressing the plate-shaped member 201 and the plate-shaped member 203 from the outside, the plate-shaped member 202 and the plate-shaped member 20 formed by the recess 206 are formed.
Insert a thin blade (usually a safety razor blade) into the gap between 3 and move the blade along the gap to cut the tissue sample 209. A thin piece having a thickness corresponding to the depth of the recess 207 is cut out. When the screw 208 is loosened and the plate member 203 is separated from the plate member 202, the cut thin piece is recessed 207.
Since it is attached to the bottom of the, peel it off with a spatula and throw it in saline.

In this method, it is necessary to remove or attach the plate-like member 203 to the plate-like member 202 every time one thin piece is cut, and it takes time to form a plurality of thin pieces.
Over time, the tissue loses its biological activity. In addition, it was difficult to make the thickness of the slice constant by this method, and it could not be applied to a very soft tissue such as the brain.

Another method conventionally used is to insert a spacer 103 having a required thickness between two safety razors 101 and 102 as shown in FIG. 2 and fix them with a clip or the like to obtain a semi-frozen tissue. Cut a block (called a block) of this combination blade, remove one of the blades,
Is taken out to obtain a sliced sample having a required thickness. In this method, even a soft tissue in a normal state such as a brain can be cut by using a block that is in a semi-frozen state to an appropriate hardness during freezing. However,
If the freezing progresses too much, it will be too hard and will not cut, so it is necessary to adjust the frozen state slightly.

Therefore, according to this method, when it is desired to obtain a large number of thin-section samples from one block, the frozen state of the tissue block changes during the repeated thin-section work, making it difficult to perform the thin-section. The number of thin slices that could be collected from the plant was at most a few, and the number was extremely limited. If it freezes too much, you can thaw it by freezing it and freezing it again.
Repeated freezing impairs the cellular properties and functions of the tissue. Even in the case of a tissue that can be sliced without being in a semi-frozen state (for example, liver), the characteristics and function of the tissue will be impaired unless the work is completed in a short time.

[0007]

DISCLOSURE OF THE INVENTION The present invention is a method for obtaining a large number of sliced samples of a constant thickness from one block of biological tissue without impairing the properties and functions of biological tissue at the cellular level. For the purpose of providing.

[0008] A second object of the present invention is to obtain a large number of slices of a constant thickness from one block of biological tissue without impairing the characteristics and function of biological tissue at the cellular level. It is the provision of equipment.

[0009]

The first object of the present invention is to provide, in the present invention, four or more thin plate-shaped blades (blades) having the same straight or curved blades at predetermined intervals by a fixing means. Using a combination of blades that are fixed so that they are parallel to each other and the blades are also parallel to each other, thinly cut a biological tissue block and insert it between the blades, each of which has a predetermined thickness and is fixed in a substantially parallel plate shape. It was achieved by a method comprising inserting an extrusion member into each gap of the blades and extruding the flakes contained between the blades.

It is preferable that the surface of the blade is coated with a hydrophobic liquid (eg liquid paraffin) or a hydrophilic liquid having a relatively high viscosity (eg polyethylene glycol).

In the present invention, the second object is a thin plate-shaped tool having four or more blades having the same straight or curved shape (hereinafter referred to as a blade) and a predetermined blade insertable between the blades. A flat plate-shaped pushing member, a blade fixing means for fixing the blades in parallel with each other, and an extruding member fixing means for fixing the extruding members substantially in parallel with each other, and the blades are formed by the extruding members. Parallel to each other with a predetermined interval that can be inserted between
Also, the blades are fixed so as to be parallel to each other, and the push-out member fixing means fixes the push-out members substantially parallel to each other at a predetermined interval so that they can be smoothly inserted between the blades. It was achieved by means of a biological tissue slicer sample preparation instrument adapted to be inserted into the.

Any number of blades may be used as long as the number of blades is four or more, but a larger number of blades provides a more uniform thickness. The size of the blade depends on the size of the object to be sliced and is not particularly limited, but usually the thickness is 0.05 to 1 mm, the depth (direction perpendicular to the blade) is 4 to 50 mm, and the length (crossing of the blade) is 10 to 200 mm. The blade spacing is determined by the desired thickness of the flakes, but is usually 0.2 to 3 mm. If the tissue is particularly soft, it is necessary to increase the interval between the blades, and for example, in the case of a thin section of the brain, it is preferably 0.8 mm or more. The blade may be linear or curved (for example, an arc). The blades of the plurality of blades are parallel to each other. It is preferable that all the blades are located in one plane perpendicular to the plane of the blade (one plane when the blades are straight).

Since each of the pushing members must be easily inserted between the blades, the thickness thereof depends on the distance between the blades and is slightly smaller than the distance between the blades, but is usually 0.15 to 5 mm. . The width of the extruding member (in the direction parallel to the blade of the blade) also depends on the width of the blade and the width of the gap, and the dimension is slightly smaller than that, but it is usually 8 to 200 mm. The length of the extruding member (direction perpendicular to the blade of the blade) is slightly larger than the depth of the blade so that the thin piece can be completely extruded, but is usually 5 to 60 mm.

It is desirable that the blade and the pushing member are made of a material that does not rust or corrode by the tissue material and does not affect the biological characteristics or function of the tissue material. Stainless steel is suitable as a material for the blade. The material of the extruding member is plastic,
Glass, other ceramics, stainless steel, brass, etc. can be used. Since the blade fixing means and the pushing member fixing means may come into contact with the tissue material, it is preferable to use the same material.

The extruding of the flakes can be carried out from the blade of the blade, but it is preferable to carry out from the opposite side of the blade.

It is preferable to carry out the operations of slicing and extruding while keeping the periphery of the device at a low temperature with ice, a cold insulating material or the like.

The method of the present invention is useful for obtaining a large number of sliced samples from a tissue block in a short time. In particular, it is also effective for obtaining a large number of sliced samples in a semi-frozen state from a very soft tofu-like tissue such as the brain. By using the method of the present invention, a portion of the tissue sample loses biological activity during work,
Alternatively, it is possible to prevent the change of the characteristic or function at the cellular level.

[0018]

In the combined blade used in the method of the present invention, four or more thin blade-shaped blades (blades) having the same straight or curved blade are parallel to each other at a predetermined interval by the fixing means and the blades are also parallel to each other. It is fixed so that
When a biological tissue block is pushed through using this combined blade, a plurality of thin slices of biological tissue corresponding to the number of blade gaps are cut off at once from the block with a thickness according to the blade interval, and the thin slice is between the blades. To position. The flat plate-shaped pushing member has a thickness that can be inserted between the blades, is fixed substantially in parallel, and can be inserted into each gap of the blades. The extruding member is inserted between the blades to extrude the thin pieces between the blades, and the thin pieces are taken out of the blade. The ejected slices of biological tissue have a thickness approximately equal to the blade spacing. A biological tissue slice having a thickness corresponding to the blade interval is obtained.

The biological tissue slicing sample preparation device of the present invention comprises 4
A thin plate-shaped blade (blade) with one or more blades with the same straight or curved shape, a flat plate-shaped extruding member with a specified thickness that can be inserted between each blade, and blade fixing that fixes the blades in parallel with each other Means and pusher member securing means for securing the pusher members substantially parallel to each other. The blades are fixed by the blade fixing means so that the respective extruding members can be inserted between them, that is, with a gap not smaller than the thickness of the extruding members, so that the blades are parallel to each other and the blades are also parallel to each other. Therefore, each extruding member can be inserted individually into the blade gap. Further, since the push-out members are fixed by the push-out member fixing means substantially parallel to each other at intervals such that they can be smoothly inserted between the blades, the push-out members can be inserted simultaneously between the blades. .

When the biological tissue block is pushed down by using the combined blade comprising the blade and the blade fixing means, a plurality of thin slices of biological tissue corresponding to the number of blade gaps are formed from the block at a thickness according to the blade spacing. Cut off,
The flakes are located between the blades. The extruding member fixed by the extruding member fixing means so that it can be inserted into each gap of the blades is inserted between the blades, and the thin pieces contained between the blades are extruded to remove the thin pieces from the blade. Can be taken out. The ejected slices of biological tissue have a thickness approximately equal to the blade spacing. A biological tissue slice having a thickness corresponding to the blade interval is obtained.

[0021]

EXAMPLES The following examples are given to further illustrate the present invention. [Example 1] Fig. 1 shows an apparatus for preparing a thin sample of biological tissue according to the present invention and used in the present invention. 1A is a plan view, FIG. 1B
Is a front view and FIG. 1C is a cross-sectional view taken along the section line XX. The biological tissue thin section sample preparation tool is composed of five blades 1 each having the same linear blade, an extrusion plate 2 having a thickness that can be inserted between each blade 1, and a blade fixing for fixing the blades 1 in parallel with each other. It comprises a member 3 and a push plate fixing member 4 for fixing the push plate 2 in parallel with each other. The blades 1 are fixed by a blade fixing member 3 so that they are parallel to each other at regular intervals and the blades are also parallel to each other. The push-out plate 2 is also fixed in parallel with each other at a constant interval by the push-out plate fixing member 4. The spacing between the blades 1 and the spacing between the pushing plates 2 are such that the pushing plates 2 are in contact with the blades 1 and are smoothly inserted therebetween.

The blade 1 is made of stainless steel having a thickness of about 0.1 mm, and the extrusion plate 2 is made of plastic having a thickness of about 0.9 mm. Therefore, the clearance of the blade 1 is also about 0.9 mm (center spacing is about 1
cm).

The five blades 1 connected by the blade fixing member 3 constitute a combined blade of five blades.
Extrusion plate 2 inserted in the gap of blade 1 as shown
Can move in the direction of the arrow in the figure and reach the position shown by the chain line in the figure.

When the biological tissue block is pushed through by using the combined blade composed of the blade 1 and the blade fixing member 3, a plurality of biological tissue thin pieces having a thickness corresponding to the interval of the blade 1 and corresponding to the number of the gaps of the blade 1 are formed. It is cut from the block at once. The sliced flakes are located in the gap of the blade 1. When the push plate 2 is inserted into each gap of the blades 1 and pushed in the direction of the arrow in the figure, the tissue slices in the gaps of the blades 1 are pushed out of the blade.
The ejected slices of biological tissue had a thickness (about 0.9 mm) approximately equal to the distance between the blades 1. In this way, a slice of biological tissue having a thickness corresponding to the distance between the blades 1 is obtained.

[0025]

According to the method of the present invention, since three or more tissue samples can be sliced simultaneously from one block of biological tissue, a large number of sliced samples can be collected in a short time, and biological tissue at the cellular level can be obtained. Multiple slices of biological tissue can be obtained without loss of properties or function. Moreover, the thickness of the obtained sliced sample is constant.

Further, when the instrument of the present invention is used, three or more tissue samples can be sliced from one block of biological tissue at the same time, so that a large number of sliced samples can be collected in a short time, and biological tissue at the cell level can be obtained. Multiple slices of biological tissue can be obtained without loss of properties or function. Moreover, the thickness of the obtained sliced sample is constant.

[Brief description of drawings]

FIG. 1 is a plan view (A), a side view (B) and a sectional view (C) showing an embodiment of the present invention.

FIG. 2 is a plan view (A) and a sectional view (B) showing a conventional method for slicing biological tissue.

FIG. 3 is an exploded perspective view (A) and a sectional view (B) showing a conventional method for slicing biological tissue.

[Explanation of symbols]

 DESCRIPTION OF SYMBOLS 1 blade 2 extrusion plate 3 blade fixing member 4 extrusion plate fixing member 101, 102 safety razor 103 spacer 104 thin piece 201 plate member 202 plate member 203 plate member 204 columnar protrusion 205 hole 206 recess 207 recess 208 screw 209 tissue sample

Claims (6)

[Claims] 1. A combination blade comprising four or more plate-shaped blades having the same straight or curved blades fixed by a fixing means so as to be parallel to each other and the blades are also parallel to each other. , Cutting a biological tissue block to a predetermined thickness, and inserting a flat plate-shaped extruding member of a predetermined thickness, which can be inserted between each of the blades, fixed substantially in parallel, into each gap of the blades. And extruding a thin slice of biological tissue in the gap between the blades. 2. The method for preparing a biological tissue sample according to claim 1, wherein the biological tissue block is in a semi-frozen state. 3. The method for preparing a biological tissue sample according to claim 1, wherein the surface of the blade is coated with a hydrophobic liquid. 4. The method for preparing a biological tissue sample according to claim 3, wherein liquid paraffin is applied as the hydrophobic liquid. 5. A plate-shaped blade having four or more blades having the same linear or curved shape, a flat plate-shaped extruding member having a predetermined thickness that can be respectively inserted between the blades, and the blades parallel to each other. The blade fixing means for fixing, and the pushing member fixing means for fixing the pushing member substantially in parallel with each other, by the cutting tool fixing means, the cutting tool is parallel to each other at a predetermined interval at which each pushing member can be inserted. , And the blades are fixed so as to be parallel to each other, and the push-out member fixing means fixes the push-out members substantially parallel to each other at predetermined intervals so that they can be smoothly inserted between the blades. Is configured to be inserted between the blades, respectively. 6. The biological tissue slicing sample preparation tool according to claim 5, wherein the blade has a linear blade, and the blade is located in a plane perpendicular to the plane of the blade.