JP2006038466A - Cutter for forming brain slice and brain slice forming method - Google Patents

Abstract

<P>PROBLEM TO BE SOLVED: To fix a brain in a stable form without using a complicated and expensive apparatus and minding the size or shape of the brain and to easily form a brain slice having a cut surface vertical to the definite axis of the brain in a short time. <P>SOLUTION: This cutter for forming the brain slice is equipped with a main body part having a brain fixing plate for receiving the brain in a stationary state and a cutting part having a pair of wall parts at the interval corresponding to the width of the brain fixing plate so as to enable the brain fixing plate to slide in its longitudinal direction. <P>COPYRIGHT: (C)2006,JPO&NCIPI

Description

  The present invention relates to a brain slice cutting apparatus and a brain slice manufacturing method.

  Conventionally, as a cutting device for brain slice preparation, for example, `` brain slicer '' for mice and rats, `` brain matrix '' etc., hollowed in the shape of brain, acrylic, stainless steel, aluminum, A brain sample is put in a block made of zinc, and slits are engraved in the block at equal intervals (1 mm, 2 mm, or 3 mm), and a fresh brain slice can be obtained simply by dropping a razor blade into the slit. Products are known (see, for example, Non-Patent Documents 1 and 2).

  In addition, there is known a tissue embedding apparatus for cutting an irregular tissue lump into a plate shape having a cross section having the same direction as a scanning plane used for CT, MRI, and PET scanning (for example, Patent Documents). 1). In this device, the tissue mass is embedded in an alginate plastic polymer using an embedding device, so that the tissue mass has a shaped outer shape and is adapted to the cross-sectional device. . In this manner, for example, tissue and organs having irregular surfaces such as brain and kidney can be reproducibly cut in any direction (see, for example, Patent Document 1).

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However, in the above-mentioned former device, since it is necessary to install and fix the brain from the curved portion side of the back of the brain in the concave portion of the mold having a substantially matching brain shape with the back of the brain reversed, for example, There are morphological abnormalities in the anteroposterior and lateral directions due to the size of the brain according to each developmental stage of various experimental animals, developmental abnormalities, etc., such as drug toxicity tests using various experimental animals A large number of mold recesses that match the shape of the brain had to be prepared in advance, and this was a burden on the preparation of experiments by researchers and workers. At the same time, if you can't find one with a matching mold recess as described above, you can't fix the brain in a mold recess in a stable manner. It was not easy to produce.
On the other hand, in the above-mentioned latter method, since the operation of embedding the tissue is essential, it is necessary to use a complicated and expensive device, and the operation method of the device by a researcher or worker, In addition, there is a problem that it takes a lot of time to prepare a brain slice.

As a result of intensive studies under such circumstances, the present inventors have arrived at the present invention.
That is, the present invention
1. A cutting device for producing a brain slice,
A main body having a brain fixing plate for receiving the brain in an upright state;
A device having a pair of wall plates (hereinafter referred to as the present invention) having a pair of wall plates at a width interval of the brain fixing plate so that the brain fixing plate can slide in the longitudinal direction of the brain fixing plate. May also be referred to as a device);
2. The pair of wall plates included in the cutting portion is a wall plate having an end side portion that can accurately and smoothly guide a straight vertical movement of a cutting member for brain cutting. Equipment of;
3. Any one of the preceding paragraphs 1 to 2, wherein the pair of wall plates included in the cutting portion are a pair of wall plates that can be installed according to a width interval of a brain fixing plate that matches a brain size. A device according to the preceding paragraph;
4). The device according to any one of the preceding items 1 to 3, wherein the brain fixing plate of the main body is a slide glass;
5. The said main-body part is equipped with the mirror plate for enabling observation of the bottom face of the brain normally placed on the surface of the said brain fixation board from upper direction, The preceding clause in any one of the preceding clauses 1 thru | or 4 characterized by the above-mentioned. apparatus;
6). The said main-body part is equipped with the illumination means for enabling observation of the bottom face of the brain normally placed on the surface of the said brain fixed board from upper direction, The preceding clause in any one of the preceding clauses 1 thru | or 5 characterized by the above-mentioned. apparatus;
7). Any one of the preceding items 1 to 6, wherein the main body includes a guide reference line that allows a portion for cutting a brain slice to be specified in a direction perpendicular to a sliding direction of the brain fixing plate. The apparatus described in the preceding paragraph;
8). The apparatus according to any one of the preceding items 1 to 7, wherein the main body portion includes a reference line corresponding to a median brain axis in the center along the longitudinal direction of the brain fixing plate;
9. A method for preparing a brain slice,
(1) a first step in which the brain is placed on the surface of the brain fixing plate of the main body provided in the brain slice preparation cutting device;
(2) a second step in which the brain fixing plate is slid in the longitudinal direction to specify a site for cutting a brain section;
(3) The cutting member for cutting the brain is a wall plate of a cutting section provided in the cutting device for brain slice preparation, and the brain fixing plate is slidable in the longitudinal direction of the brain fixing plate. A third step (4) of obtaining a brain cut surface by moving up and down linearly along the end sides of a pair of wall plates with a width interval of (4) Repeating the second step and the third step, A method comprising a fourth step of cutting out (hereinafter, also referred to as the present invention method);
10. 10. The method according to item 9 above, wherein the surface of the brain fixing plate in the first step is the surface of the brain fixing plate after the adhesive material having fluidity is spread thinly in advance.
11. The method according to item 10 above, wherein the fixing material is an agar solution or a gelatin solution;
12 In the first step, the brain midline axis existing on the bottom surface of the brain is set to a reference line corresponding to the brain midline axis provided in the center along the longitudinal direction of the brain fixing plate of the main body part included in the brain slice preparation cutting device. The method according to any one of the preceding items 8 to 11, wherein the brain is placed on the surface of the brain fixing plate.
Etc. are provided.

  The device of the present invention and the method of the present invention can fix the brain in a stable manner without having to use a complicated and expensive device, and without worrying about the size or shape of the brain. It has become possible to easily produce a brain section having a vertical cut surface in a short time.

The present invention is described in detail below.
The present invention relates to a brain slice cutting apparatus and a brain slice manufacturing method.
First, the device according to the present invention will be described. The device according to the present invention is a cutting device for preparing a brain slice, and includes a main body portion having a brain fixing plate that receives the brain in an in-place state, and the brain fixing plate is And a cutting section having a pair of wall plates at a width interval of the brain fixing plate so as to be slidable in the longitudinal direction.

  The main body has a brain fixing plate that receives the brain in an upright state on the slide surface of the main body. The back of the brain is rounded and curved, whereas the bottom of the brain forms a flat surface from the front end to the back end, and the brain is stable on the plane with the bottom of the brain as the reference plane. It is possible to put. Here, the “normal position” means a state in which the brain is stably placed on a plane with the brain bottom as a reference plane.

  The brain fixing plate has a width that matches the size of the brain in the short direction, and has a length that is suitable for sliding on the slide surface portion of the main body in the longitudinal direction. Of course, the size and shape of the brains of various experimental animals have no effect on the sliding movement.

  The slide surface part and the brain fixing plate of the main body part are, for example, in order to make it possible to observe the bottom surface of the brain placed on the surface of the brain fixing plate, as described later, from above via a mirror plate or the like. It is preferable that the member is made of a highly transparent material such as acrylic or glass. More specifically, as a preferable brain fixation plate, a slide glass and the like can be mentioned.

  The cutting part has a pair of wall plates at a width interval of the brain fixing plate. The wall plates are vertically installed in a pair of parallel states on the slide surface portion of the main body according to the width interval of the brain fixing plate having a width matching the size of the brain. Further, the pair of wall plates may be fixed on the slide surface portion of the main body portion, but after being slid according to the width interval of the brain fixing plate having a width that matches the size of the brain. It may be a pair of wallboards that can be installed having means that can be fixed.

  The pair of wall plates included in the cutting portion has end side portions that can accurately and smoothly guide the linear vertical movement of a cutting member (cutting blade) for brain cutting such as a razor and a knife, for example. Therefore, the cutting member (cutting blade) can be correctly oriented due to the presence of the end side portion, and thus the correct cut surface of the brain slice can be accurately obtained.

In the device according to the present invention, it is preferable that the main body portion includes a mirror plate for allowing the bottom surface of the brain placed on the surface of the brain fixing plate to be observed from above. Since such a mirror plate is for specifying a site for cutting a brain section, it is preferable that the mirror plate has a flat surface with little distortion and turbidity.
The structure of the bottom surface of the brain (that is, the bottom surface of the brain placed on the surface of the brain fixing plate) projected above the slide surface portion of the main body and the mirror plate installed at the lower part (lower layer) of the brain fixing plate is shown above. While observing from the guide reference line as will be described later (that is, a guide reference line provided in a direction perpendicular to the sliding direction of the brain fixing plate, which can specify a site for cutting the brain slice: A site for cutting a brain section can be specified based on the guide reference lines (1) to (7) in FIG.

In the device according to the present invention, the main body preferably includes an illuminating unit for allowing the bottom surface of the brain placed on the surface of the brain fixing plate to be observed from above.
An illuminating means for giving light to the inside (preferably the lower part (lower layer)) of the main body, for example, a shakassen (a fluorescent plate for viewing the film with transmitted light) is placed in front of the light source to place the basal surface of the brain. Is properly illuminated and the image is properly projected in the mirror, so that the brain slice can be cut more accurately. Incidentally, when the illumination is strong, the amount of light entering the eye increases, and it may be difficult to cut the brain slice. Therefore, it is preferable to cover the light source with ground glass and reduce the amount of light.

  In the device according to the present invention, it is preferable that the main body portion includes a guide reference line that enables a portion for cutting a brain section to be specified in a direction perpendicular to a sliding direction of the brain fixing plate. As the guide reference line, for example, the presence of an end side portion that can accurately and smoothly guide the linear up-and-down movement of a cutting member (cutting blade) for brain cutting on a pair of wall plates included in the cutting portion A single thread along the position can be mentioned. The thread is a thread for accurately identifying a portion for cutting a brain section, which is projected on a mirror plate, and preferably has a thickness that does not interfere with observation and is easy to see. Further, the guide reference line may be formed by scratching a thin line on the main body and coloring it.

  In the device according to the present invention, it is preferable that the main body portion includes a reference line corresponding to the median brain axis in the center along the longitudinal direction of the brain fixing plate. As the reference line, for example, a single thread existing at an equal distance from a pair of wall plates included in the cutting portion can be cited. The thread is a thread for accurately identifying a portion for cutting a brain section, which is projected on a mirror plate, and preferably has a thickness that does not interfere with observation and is easy to see. Further, the reference line may be formed by scratching the main body portion with a fine line and coloring it.

  Next, the method of the present invention will be described. The method of the present invention is a method for producing a brain slice, and (1) the brain is placed on the surface of the brain fixing plate of the main body provided in the brain slice producing cutting device. A first step, (2) a second step of slidably moving the brain fixing plate in the longitudinal direction to specify a site for brain slice cutting, and (3) a cutting member for brain cutting for preparing a brain slice. A wall plate of a cutting unit included in the cutting device, and along the end side portion of the wall plate which is a pair at a width interval of the brain fixing plate so that the brain fixing plate can slide in the longitudinal direction of the brain fixing plate. A fourth step of obtaining a brain section by moving up and down in a straight line, and (4) a fourth step of cutting out a brain section by repeating the second step and the third step (that is, , The method of the present invention) and the like are also provided.

In the method of the present invention, the surface of the brain fixing plate in the first step is preferably the surface of the brain fixing plate after the adhesive material having fluidity is spread thinly in advance. As the fixing material, a material that is transparent and can be easily cut, and has low permeability to the brain and little influence on the brain is preferable. Specific examples include an agar solution or a gelatin solution. When an agar solution is used as the fixing material, an agar solution of about 3 to 3.5% by weight is preferable.
As a specific method for preparing the agar solution, for example, 1 g of agar powder manufactured by Nacalai Tesque is weighed, and 30 ml of distilled water is added thereto. The resulting mixture is heated to about 70 ° C. and all agar powder is dissolved while stirring. After complete dissolution, the temperature in the solution is lowered to about 45 ° C., and this is thinly spread on the surface of the brain fixation plate by about 1 to 2 ml. The brain is then quickly placed on the surface of the brain fixation plate. In this case, as will be described later, the midline axis of the brain existing on the bottom surface of the brain corresponds to the midline axis of the brain provided at the center along the longitudinal direction of the brain fixing plate of the main body part provided in the brain slice preparation cutting device. It is preferable that the brain is placed and fixed on the surface of the brain fixing plate according to the reference line.

  In the method of the present invention, in the first step, the midline axis of the brain existing on the bottom surface of the brain is the midline axis of the brain provided in the center along the longitudinal direction of the brain fixing plate of the main body part provided in the brain slice preparation cutting device. The brain may be placed on the surface of the brain fixing plate in accordance with the corresponding reference line.

  In the method of the present invention, in the second step, the brain fixing plate may be slid in the longitudinal direction to specify a site for brain section cutting. Specifically, for example, when the main body portion includes a mirror plate for allowing the bottom surface of the brain placed on the surface of the brain fixation plate to be observed from above, the slide surface portion of the main body portion and the brain fixation While observing the structure of the bottom surface of the brain (that is, the bottom surface of the brain placed on the surface of the brain fixing plate) projected on the mirror plate installed at the bottom (lower layer) of the plate from above, the guide reference line (That is, a guide reference line provided in a direction perpendicular to the sliding direction of the brain fixing plate and capable of specifying a site for cutting a brain slice: for example, (1) to (7) in FIG. A site for brain section cutting can be identified based on the guide line shown).

  In the method of the present invention, in the third step, the cutting member for cutting the brain is a wall plate of a cutting section provided in the cutting device for preparing a brain slice, and the brain fixing plate is slidable in the longitudinal direction of the brain fixing plate The brain cut surface may be obtained by moving up and down linearly along the end sides of the pair of wall plates at a width interval of the brain fixing plate. Specifically, for example, the pair of wall plates included in the cutting portion is an end that can accurately and smoothly guide the linear vertical movement of a cutting member (cutting blade) for brain cutting such as a razor or a knife. It has a side portion, and the presence of the end side portion enables the cutting member (cutting blade) to be correctly oriented, whereby the correct cut surface of the brain slice can be obtained accurately.

  In the method of the present invention, in the fourth step, the brain section may be cut out by repeating the second step and the third step. By appropriately adjusting the slide movable width in the longitudinal direction of the brain fixing plate in the second step, it is possible to cut out a brain section of a desired thickness, and a site for cutting a desired brain section It is also possible to cut out brain slices.

  Hereinafter, the present invention will be described in more detail with reference to examples. However, the present invention is not limited to these examples.

Example 1
One embodiment of the device of the present invention and the method of the present invention is illustrated as shown in FIGS.

First, an experimental animal to which a chemical substance has been administered is euthanized, the head skin of the experimental animal is incised left and right from the midline, the skull is excised using scissors, and then the brain is carefully removed. The isolated brain is used as it is or fixed with a formalin solution or the like. The size of the extracted brain varies depending on the age, but it is about 21 mm long × 15 mm wide × 10 mm thick in an 8-week-old rat. In this way, a brain for preparing a brain section was prepared.
Next, a slide glass (76.1 mm long × 26 mm wide × 1.2 mm thick) which is a brain fixation plate removed from the main body (148 mm long × 148 mm wide × 27.6 mm thick) ((4) in FIG. 3) ), A small amount (about 1 to 2 ml) of a dissolved agar solution (about 45 ° C.) is spread thinly, and then the median axis of the brain observed at the bottom of the brain ((8) in the left figure in FIG. 1) The brain (median) is fixed in an upright state (upper left in FIG. 4) so as to coincide with a reference line ((5) in FIG. 3) existing in the center along the longitudinal direction of the fixing plate. (Upper right figure in FIG. 4).
After the brain fixing plate is placed on the main body, the target structure of the brain bottom (shown in FIG. 1) projected on the mirror plate (89 mm long × 89 mm wide × 2 mm thick) ((2) in FIG. 3) of the main body (See the figure on the right) so that the end side of the wall plate (26.6 mm length × 13.1 mm width × 2.2 mm thickness) ((3) in FIG. 3) of the cutting unit provided in the device of the present invention is aligned. , Slide the brain fixation plate in its longitudinal direction. In this way, the region for cutting the brain section ((1) to (7) in FIG. 1) is specified.
A pair of razors, which are cutting members for brain cutting, are spaced apart by the width of the brain fixing plate so that the brain fixing plate is slidable in the longitudinal direction of the brain fixing plate. While closely contacting the end side of the wall plate ((3) in FIG. 3), it is moved up and down in a straight line with the pressure on the brain kept small (lower left figure in FIG. 4). (1) to (7)) in FIG. 2 are obtained.
Furthermore, by repeating such an operation, a desired brain slice is easily cut out in a short time.

  The device of the present invention and the method of the present invention can fix the brain in a stable manner without having to use a complicated and expensive device, and without worrying about the size or shape of the brain. It has become possible to easily produce a brain section having a vertical cut surface in a short time.

FIG. 1 is a diagram showing a cut site of a brain section of a rat brain. The figure on the left shows the cut site of the brain section from the upper side of the brain, and the figure on the right shows the cut site of the brain section from the lower side of the brain. In addition, (1) to (7) in the figure represent guide reference lines that can specify a site for cutting a brain slice in a direction perpendicular to the sliding direction of the brain fixation plate. (8) represents a reference line corresponding to the median brain axis. FIG. 2 is a diagram showing a cut site of a brain section of a rat brain. In addition, (1)-(7) in the figure represents a guide reference line that makes it possible to specify a part for cutting a brain slice in a direction perpendicular to the sliding direction of the brain fixing plate. It is the figure which showed the structure of an example of this invention apparatus. The figure includes three drawings in the order of a top view in the upper stage, a side view in the lower left stage, and a front view in the lower right stage. In the figure, (1) represents a slide surface portion, (2) represents a mirror plate, (3) represents a wall plate, (4) represents a brain fixation plate, (5) is a guide reference line (that is, a guide reference line that is provided in a direction perpendicular to the sliding direction of the brain fixing plate and that can specify a region for cutting a brain slice) and a guide reference line (that is, (Guide reference line enabling identification of a part for cutting a brain section), and (6) represents an illumination device. It is the figure (photograph) which showed the outline | summary of an example of this invention apparatus and this invention method. The upper left is a photograph showing the main body having a brain fixing plate that receives the brain in an in-place state, and the brain placed on the surface of the brain fixing plate, and the upper right is the surface of the brain fixing plate. It is a photograph showing a whole view of the device of the present invention with the brain placed on top, and the lower left is a wall plate of a cutting section provided in the cutting device for preparing a brain section with a cutting member for cutting the brain. A step of obtaining a brain cut surface by linearly moving up and down along the end sides of a pair of wall plates at a width interval of the brain fixed plate so that the fixed plate can slide in the longitudinal direction of the brain fixed plate The lower right is a photograph showing a brain slice sliced at the target site.

Explanation of symbols

(1) Slide surface portion (2) Mirror plate (3) Wall plate (4) Brain fixation plate (5) Guide reference line (ie, brain slice cutting provided in a direction perpendicular to the slide direction of the brain fixation plate) Guidance reference line that enables identification of a part for the brain) and guidance reference line (that is, guidance reference line that enables identification of the part for cutting the brain slice)
(6) Lighting device

Claims (12)

A cutting device for producing a brain slice,
A main body having a brain fixing plate for receiving the brain in an upright state;
An apparatus comprising: a cutting unit having a pair of wall plates at a width interval of the brain fixing plate so that the brain fixing plate can slide in the longitudinal direction of the brain fixing plate.   The pair of wall plates included in the cutting portion are wall plates having end side portions capable of accurately and smoothly guiding a straight vertical movement of a cutting member for brain cutting. The device described.   The pair of wall plates included in the cutting section are a pair of wall plates that can be installed according to the width interval of the brain fixation plate that matches the size of the brain. An apparatus according to any of the claims.   The device according to any one of claims 1 to 3, wherein the brain fixing plate of the main body is a glass slide.   The said main-body part is equipped with the mirror board for enabling observation of the bottom face of the brain arranged on the surface of the said brain fixed board from upper direction, The Claim 1 thru | or 4 characterized by the above-mentioned. The device described.   The said main-body part is equipped with the illumination means for enabling observation of the bottom face of the brain normally placed on the surface of the said brain fixed board from upper direction, The Claim 1 thru | or 5 characterized by the above-mentioned. The device described.   The said main-body part is equipped with the guide reference line which can specify the site | part for brain slice cutting | disconnection in the orthogonal | vertical direction with respect to the sliding direction of the said brain fixing board. The device of claim 1.   The apparatus according to claim 1, wherein the main body includes a reference line corresponding to a median brain axis at a center along a longitudinal direction of the brain fixing plate. A method for preparing a brain slice,
(1) a first step in which the brain is placed on the surface of the brain fixing plate of the main body part included in the brain slice preparation cutting device;
(2) a second step of sliding the brain fixation plate in the longitudinal direction to specify a site for cutting a brain section;
(3) A cutting member for cutting the brain is a wall plate of a cutting section provided in the cutting device for preparing a brain section, and the brain fixing plate is slidable in the longitudinal direction of the brain fixing plate. A third step (4) of obtaining a brain cut surface by moving up and down linearly along the end sides of a pair of wall plates with a width interval of (4) Repeating the second step and the third step, A method comprising a fourth step of cutting out.   The method according to claim 9, wherein the surface of the brain fixation plate in the first step is the surface of the brain fixation plate after the adhesive material having fluidity is spread thinly in advance.   The method according to claim 10, wherein the fixing material is an agar solution or a gelatin solution. In the first step, the brain midline axis existing on the bottom surface of the brain is set to a reference line corresponding to the brain midline axis provided in the center along the longitudinal direction of the brain fixing plate of the main body part included in the brain slice preparation cutting device. The method according to claim 8, wherein the brain is placed on the surface of the brain fixing plate.

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