JP6982859B2 - Method for manufacturing primary embedding block for otolith observation, specimen block for otolith observation, and otolith sample piece - Google Patents

Description

The present invention relates to a primary embedding block for otolith observation for forming an otolith sample piece for observing a cross section of a fish otolith, a sample block for otolith observation, and a method for producing an otolith sample piece.

Otoliths made of calcium carbonate are present in the lower semicircular canals of fish. Since this otolith forms a ring with growth like an annual ring of a tree centered on the nucleus, the age of the fish can be grasped by counting these. In addition, when the ring of the otolith is magnified and observed, it is possible to observe a finer ring called a sun ring, which is formed every day. Using these otoliths, it is possible to carry out a wide range of fish research, such as assessing the age of fish, analyzing resources, and observing the growth process of fry.

In the analysis of the otolith, an observation cross section with the core of the otolith and the ring surrounding it exposed is formed by polishing the otolith. Conventionally, as a method of polishing a sample to form a specific observation cross section, for example, in Non-Patent Document 1, for example, in Non-Patent Document 1, an otolith as a sample is embedded in an epoxy resin and then the epoxy resin is cured to form the otolith package. It is described that the buried block is cut near the center of the otolith and then polished to form a sample piece with an observed cross section that exposes the core of the otolith and the ring surrounding it.

On the other hand, in the study of fish using otoliths, it is necessary to analyze hundreds to thousands of otoliths in order to improve the analysis accuracy. Conventionally, in order to efficiently form an observation cross section of a large number of otoliths, for example, a large number of otoliths are embedded in a transparent resin or the like and cured, and a large number of otoliths embedded in the resin are used. A method of efficiently forming an observed cross section of an otolith by polishing all at once is performed.

Bulletin of Faculty of Fisheries, Kagoshima University-Memoirs of Faculty of Fisheries Kagoshima University, 52: 51-56

However, conventionally, when embedding an otolith as an observation sample in a resin or the like, there is a problem that it is difficult to position the embedding position. For this reason, when the otolith is fixed, its inclination and fixing position are not constant, and it is difficult to accurately form an observation cross section in which the core of the otolith and the ring around it can be observed as a whole. ..

The present invention has been made in view of the above-mentioned situation, and is a primary embedding block for otolith observation, an ear, which can easily and accurately form an observation cross section including an otolith core of a fish. It is an object of the present invention to provide a method for manufacturing a sample block for observing stones and a sample piece of otolith.

That is, the primary embedding block for otolith observation of the present invention has the following configuration.
A fish otolith sample, a base material having a flat sample fixing surface for fixing the fish otolith sample, and an embedding member for fixing the otolith sample to the sample fixing surface of the base material are provided. A positioning index indicating the position of the nucleus of the otolith sample is formed on the sample fixing surface, and the positioning index is an intersection line consisting of two straight lines intersecting each other at the center point of the nucleus. It is characterized by being formed at a position separated from the center point and further provided with a processing index line indicating the distance to the center point along one of the straight lines of the crossing line.

According to the present invention, by forming a positioning index indicating the position of the core of the otolith sample on the sample fixing surface of the primary embedding block for otolith observation, the position of the core of the otolith can be reliably grasped and the otolith The sample can be fixed to the sample fixing surface.

Further, in the present invention, the auxiliary index line has one end extending toward the center point and one end away from the center point along a direction inclined at a predetermined angle with respect to the intersection line. It is characterized by being further equipped with.

The specimen block for otolith observation of the present invention has the following configuration.
Said otolith observation primary embedded block, the otolith sample is a transparent covering member covering the otolith observation primary embedded block fixed, characterized in that it consists of.

The method for producing an otolith sample piece of the present invention has the following constitution.
A method for producing an otolith sample piece using the otolith observation sample block, wherein the otolith sample is placed on the sample fixing surface so that the core of the otolith sample coincides with the center point of the intersection line. The otolith fixing step of fixing to the otolith, the covering step of covering the primary otolith observation block to which the otolith sample is fixed with a transparent covering member to form the otolith observation specimen block, and the intersection line. It has at least a polishing step of scraping the otolith observation specimen block from two directions toward the core of the otolith sample along one of the straight lines forming the above, and the polishing step includes a processing index line and an auxiliary. It is characterized in that the sample block for otolith observation is scraped to a position where a part of the core of the otolith sample is exposed while grasping the distance to the core of the otolith sample with reference to the index line.

According to the method for producing an otolith observation primary embedding block, an otolith observation specimen block, and an otolith sample piece of the present invention, an observation cross section including a fish otolith core can be easily and accurately formed. Will be possible.

It is a perspective view and the side view which shows the primary embedding block for otolith observation of one Embodiment of this invention in the state which the otolith (otolith sample) is fixed. It is a perspective view and the side view which shows the primary embedding block for otolith observation of another embodiment of this invention in the state which the otolith (otolith sample) is fixed. It is a top view which shows the crossing line, the auxiliary index line, and the auxiliary index line formed on the sample fixing surface. It is a perspective view which shows the specimen block for otolith observation of one Embodiment of this invention. It is explanatory drawing which shows the manufacturing process of the specimen block for otolith observation. It is a perspective view which shows an example of an otolith sample piece.

Hereinafter, a method for manufacturing an otolith observation primary embedding block, an otolith observation specimen block, and an otolith sample piece according to an embodiment of the present invention will be described with reference to the drawings. It should be noted that each of the embodiments shown below is specifically described in order to better understand the gist of the invention, and is not limited to the present invention unless otherwise specified. In addition, the drawings used in the following description may be shown by enlarging the main parts for convenience in order to make the features of the present invention easy to understand, and the dimensional ratios of each component are the same as the actual ones. It is not always the case.

FIG. 1A is a perspective view showing a primary embedding block for otolith observation of the present invention in a state where an otolith (otolith sample) is fixed. Further, FIG. 1B is a side view taken along the Y-axis direction of FIG. 1A. Further, FIG. 1 (c) is a side view of FIG. 1 (a) along the X-axis direction.
The primary embedding block 10 for otolith observation includes a base material 11 having a flat sample fixing surface 11a for fixing the otolith sample S. In the present embodiment, the base material 11 is a rectangular parallelepiped, and any one surface is a sample fixing surface 11a. The base material 11 is made of a material that can be easily polished, for example, a resin material. Further, the resin material needs to be formed of a transparent (light-transmitting) resin material.

On the sample fixing surface 11a, an intersection line (virtual intersection line) 13 consisting of two straight lines 13a and 13b indicating the center point C indicating the position of the otolith nucleus SC in the center of the otolith sample S is formed. .. Such an intersection line 13 is an index indicating the position of the axis line (minor axis, major axis) of the otolith sample S.
In the present embodiment, the crossing line 13 is composed of two straight lines a and 13b orthogonal to each other along the sample fixing surface 11a. In the following description, the axis parallel to the straight line 13a is referred to as the X axis, the axis parallel to the straight line 13b is referred to as the Y axis, and the X axis and the axis perpendicular to the Y axis are referred to as the Z axis. Such an intersection line 13 is composed of, for example, ink, for example, toner.

In the otolith sample S, the otolith sample S is the sample fixing surface 11a so that the otolith nucleus SC at the center of the otolith sample S coincides with the center point C of the intersection line 13 formed on the sample fixing surface 11a. Is fixed to. The otolith nucleus SC can be easily recognized as a black spot, for example, by transmitting light through the otolith sample S. Therefore, the base material 11 uses a light-transmitting resin.
The otolith sample S and the sample fixing surface 11a may be fixed by using, for example, an ultraviolet curable resin. This makes it possible to quickly fix the otolith sample S to the sample fixing surface 11a.

Further, on the sample fixing surface 11a, one of the two straight lines 13a, 13b, that is, in the present embodiment, the machining index lines 14a, 14b, 14c, 14d arranged at predetermined intervals along the Y axis. Is formed. The machining index lines 14a to 14d indicate the distance width along the Y axis from the center point C of the crossing line 13, and in the present embodiment, the machining index line 14a is from the center point C of the crossing line 13 to the Y axis. The machining index line 14b is located at a position of +0.5 mm along the Y-axis from the center point C of the crossing line 13 at a position of +0.5 mm, and the machining index line 14c is located at a position of +1.0 mm along the Y-axis from the center point C of the crossing line 13. A machining index line 14d is formed at a position of −0.5 mm along the line of the crossing line 13 at a position of −1.0 mm along the Y axis from the center point C of the crossing line 13. The length of the processing index lines 14a to 14d itself is formed to be 1.0 mm in the present embodiment. These processing index lines 14a to 14d are composed of, for example, ink.

Further, the sample fixing surface 11a has an auxiliary index having one end extending toward the center point C and one end at a position separated from the center point C along a direction inclined at a predetermined angle with respect to the intersection line 13. Lines 15a to 15d are formed. In the present embodiment, the auxiliary index lines 15a to 15d extend at an angle of 45 ° with respect to the crossing line 13, for example. The length of the auxiliary index lines 15a to 15d itself is formed to be 1.0 mm in this embodiment. These auxiliary index lines 15a to 15d are composed of, for example, ink.

FIG. 3 shows the crossing lines 13, the processing index lines 14a to 14d, and the auxiliary index lines 15a to 15d formed on the sample fixing surface 11a as described above. The crossing line 13 is a minute line surrounded by one end of the crossing line 13 and one end of the auxiliary index lines 15a to 15d as a virtual line without forming the vicinity of the center point C as an actual line as shown in FIG. The circular region may be set as the center point C. Further, the auxiliary index lines 15a to 15d are also virtual lines in the vicinity of the center point C.

The otolith sample S has a flat elliptical shape, and one elliptical plane is fixed to the sample fixing surface 11a. Further, in the embodiment shown in FIG. 1, the otolith sample S is fixed to the sample fixing surface 11a so that the long axis of the elliptical otolith sample S is along the straight line 13a of the crossing line 13, that is, the X axis.

As in the embodiment shown in FIGS. 2A and 2B, the otolith sample has the short axis of the elliptical otolith sample S along the straight line 13b of the crossing line 13, that is, along the X axis. S can also be fixed to the sample fixing surface 11a.

Next, the specimen block for otolith observation of the present invention will be described.
FIG. 4 is a perspective view showing a specimen block for otolith observation according to an embodiment of the present invention.
The otolith observation specimen block 20 is covered with the otolith observation primary embedding block 10 shown in FIG. 1 and the otolith sample S fixed to the sample fixing surface 11a of the otolith observation primary embedding block 10. It has a member 21.

The covering member 21 is formed, for example, into a cylindrical outer shape. The specimen block 20 for otolith observation can easily observe the crossing lines 13, the processing index lines 14a to 14d, and the auxiliary index lines 15a to 15d formed on the sample fixing surface 11a from the outside.

Next, a method for producing an otolith sample piece using the otolith observation specimen block 20 will be described with reference to FIGS. 1 and 5, including the operation of the otolith observation primary embedding block 10.
When forming an otolith sample piece in which the sample cross section including the otolith nucleus SC of the otolith sample S is exposed by the method for producing an otolith sample piece of the present invention, first, the primary for otolith observation of the present invention is formed. The embedding block 10 is prepared. Then, a small amount of curable resin is formed in the vicinity of the center point C of the crossing line 13 formed on the sample fixing surface 11a of the primary embedding block 10 for otolith observation.

Next, the otolith sample S is placed on the sample fixing surface 11a and irradiated with light transmitted through the otolith sample S.
For example, the otolith nucleus SC existing in the otolith sample S visually recognized as a black spot is confirmed. At this time, it is preferable to magnify and observe using an optical microscope or the like.

Then, when the otolith nucleus SC is confirmed, the otolith sample S is finely moved on the sample fixing surface 11a so that the otolith nucleus SC coincides with the center point C of the intersection line 13.
Further, the otolith sample S is rotated so that the long axis of the elliptical otolith sample S coincides with the straight line 13a (X axis) or the straight line 13b (Y axis) forming the crossing line 13.
Then, the curable resin is cured, and the otolith sample S is fixed to the sample fixing surface 11a of the primary embedding block 10 for otolith observation (otolith fixing step).

In this way, by forming an intersection line 13 composed of two straight lines 13a and 13b intersecting each other at the center point C on the sample fixing surface 11a of the primary embedding block 10 for otolith observation, the ears of the otolith sample S are formed. The stone core SC is located at a position overlapping the center point C of the crossing line 13, and the direction of the long axis of the elliptical earstone sample S forms the crossing line 13 as a straight line 13a (X-axis) or a straight line 13b (Y-axis). The earstone sample S can be fixed to the sample fixing surface 11a so that it can be easily grasped that any of the above is met.

Next, as shown in FIG. 5, the otolith observation primary embedding block 10 to which the otolith sample S is fixed is immersed in the transparent uncured resin material liquid Q constituting the covering member 21. Then, the resin material liquid Q is cured, and the otolith sample S fixed to the sample fixing surface 11a of the otolith observation primary embedding block 10 and the otolith observation primary embedding block 10 and the otolith sample. A covering member 21 for covering the primary otolith observation block 10 to which S is fixed and an otolith observation specimen block 20 composed of the covering member 21 are formed (covering step).

As the transparent resin material constituting the covering member 21, the same resin material as that constituting the base material 11 of the primary embedding block 10 for otolith observation is used. Further, examples of the transparent resin material constituting the covering member 21 include an ultraviolet curable resin, a thermosetting resin, a photocurable resin, and the like.

Next, the otolith observation specimen block 20 is placed on the polishing device, and the otolith sample S is placed in the ear from two directions toward the otolith nucleus SC along the straight line 13b forming the crossing line 13, that is, the Y axis. The otolith observation specimen block 20 is polished (polishing step).

At this time, by referring to the processing index lines 14a to 14d, the distance of the otolith sample S to the otolith nucleus SC can be easily grasped. For example, between the machining index line 14b and the machining index line 14a, and between the machining index line 14d and the machining index line 14c, the polished surface is at least 0.5 mm away from the otolith core SC. Therefore, high-speed polishing can be performed at a high polishing rate.

Then, when the surface to be polished reaches the positions of the machining index line 14a and the machining index line 14c, polishing is performed at a low polishing rate, and the otolith core SC and its surroundings are not scraped off. The observed cross section of the entire surrounding ring can be easily and reliably formed.

In the polishing after the positions of the machining index line 14a and the machining index line 14c, the auxiliary index lines 15a to 15d tilted at an angle of 45 ° with respect to the crossing line 13, the straight line 13a (X axis), and the straight line 13b (Y). By polishing while checking the distance to the shaft), the distance from the polished surface to the otolith nucleus SC can be accurately grasped by the triangular function, and the otolith nucleus SC will be scraped off even more reliably. The target observation cross section of the otolith sample S can be formed without any problem.
Further, when the otolith sample S is large or the like, the otolith observation sample block 20 is placed on a cutting device before the polishing step, for example, along the X-axis direction, the processing index line 14a and the processing index line. The otolith observation specimen block 20 may be cut from both sides at the position 14c.

An example of the otolith sample piece thus obtained is shown in FIG.
In the otolith sample piece 30 shown in FIG. 6, the otolith sample block 20 (see FIG. 4) is polished from both sides along the X-axis to form a disk shape.

Although some embodiments of the present invention have been described above, these embodiments are presented as examples and are not intended to limit the scope of the invention. These embodiments can be implemented in various other embodiments, and various omissions, replacements, and changes can be made without departing from the gist of the invention. These embodiments and variations thereof are included in the scope of the invention described in the claims and the equivalent scope thereof, as are included in the scope and gist of the invention.

For example, in the above-described embodiment, the crossing line is formed as the positioning index of the nucleus, but in addition to this, for example, a line serving as an index of the center point and the major axis or the minor axis, the center point and the major axis, and the short axis. A line, a center point, a circular region, etc., which are indicators of the axis, can be used as the positioning index of the nucleus, and the specific form of the positioning index of the nucleus is not limited.
Further, in the above-described embodiment, the shape of the primary embedding block for otolith observation is a rectangular parallelepiped, but in addition to this, various shapes such as a hexagonal plate shape and a disk shape can be used. It is also possible to collectively mold a plurality of primary otolith observation blocks by using a mold capable of molding a plurality of otolith observation primary embedding blocks. This makes it possible to efficiently form observation cross sections of a large number of otolith samples by fixing a large number of otolith samples together to the primary otolith observation block and covering it with a covering member. become.
Further, in the above-described embodiment, the specimen block 20 for otolith observation is formed in a cylindrical shape, but other than this, various shapes such as a cube shape and a hexagonal columnar shape can be formed.
Further, by applying the configuration of the primary otolith observation block of the present invention, for example, hard tissues of other organisms (bones, shells, otoliths of mollusks, pearls, etc.) can be embedded. In that case, the direction of polishing and the position during polishing can be determined by the crossing line, the processing index line, and the auxiliary index line for the observation target corresponding to the nucleus of each organism.

10 ... Primary embedding block for otolith observation 11 ... Base material 11a ... Sample fixing surface 13 ... Crossing line 13a, 13b ... Straight line 14a-14d ... Processing index line 15a-15d ... Auxiliary index line 20 ... otolith observation specimen block 21 ... Covering member C ... Center point S ... Otolith sample (otolith)
SC ... Otolith core Q ... Resin material liquid

Claims (4)

A fish otolith sample, a base material having a flat sample fixing surface for fixing the fish otolith sample, and an embedding member for fixing the otolith sample to the sample fixing surface of the base material are provided. ,
A positioning index indicating the position of the nucleus of the otolith sample is formed on the sample fixing surface.
The positioning index is an intersection line consisting of two straight lines intersecting each other at the center point of the nucleus.
A primary embedding block for otolith observation, which is formed at a position away from the center point and further includes a processing index line indicating a distance to the center point along one of the straight lines of the crossing line. .. Further provided with an auxiliary index line having one end extending toward the center point and one end distant from the center point along a direction inclined at a predetermined angle with respect to the crossing line. The primary embedding block for otolith observation according to claim 1. The otolith observation according to claim 2, comprising the primary otolith observation block for otolith observation and a transparent covering member covering the otolith observation primary embedding block to which the otolith sample is fixed. Specimen block for. A method for producing an otolith sample piece using the otolith observation specimen block according to claim 3.
An otolith fixing step of fixing the otolith sample to the sample fixing surface so that the core of the otolith sample coincides with the center point of the crossing line.
A covering step of covering the primary otolith observation block to which the otolith sample is fixed with a transparent covering member to form the otolith observation specimen block.
It has at least a polishing step of scraping the otolith observation sample block from two directions toward the core of the otolith sample along one straight line forming the crossing line.
In the polishing step, while grasping the distance to the core of the otolith sample with reference to the processing index line and the auxiliary index line, the sample for otolith observation is reached to a position where a part of the core of the otolith sample is exposed. A method for producing an otolith sample piece, which is characterized by scraping a block.

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