JP2019066442A - Primary embedding block for observing otolith, sample block for observing otolith, and method of preparing otolith sample piece - Google Patents

Abstract

To provide a primary embedding block for observing an otolith, which allows for easily and accurately producing an observation cross-section that includes a core of a fish otolith, a sample block for observing an otolith, and a method of preparing an otolith sample piece.SOLUTION: An embedding block comprises a substrate 11 having a flat sample fixing surface 11a for a fish otolith sample S to be fixed thereon, the sample fixing surface 11a having a cross-line (virtual cross-line) 13 consisting of two straight lines 13a, 13b formed thereon, which shows a center point that acts as an indicator for positioning a core SC of the otolith sample S.SELECTED DRAWING: Figure 1

Description

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

  In the lower brain area of the fish, there are otoliths made of calcium carbonate in the three semicircular canals. Since this otolith forms a ring with growth like a tree annual ring centering on a nucleus, the age of the fish can be grasped by counting these. In addition, when the otolith ring is enlarged and observed, it is possible to observe a more detailed ring formed every day called a sun ring. Using these otoliths, it is possible to conduct a wide range of fish research such as assessment of fish age, resource analysis, and observation of juvenile growth processes.

  In the analysis of otoliths, the observation cross section in which the ring covering the otolith nucleus and its periphery is 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, an epoxy resin is embedded with otolith that is the sample, and then the epoxy resin is cured, and the otolith capsule is formed. It is described to form a specimen having an observation cross section in which the ring covering the otolith nucleus and its periphery is exposed by cutting the buried block near the center of the otolith and then polishing it.

  On the other hand, to study fish using otoliths, it is necessary to analyze hundreds to thousands of otoliths in order to improve analysis accuracy. Conventionally, in order to efficiently form a 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 a cured resin and a large number of otoliths embedded in the resin are used. The method of forming the observation cross-section of otolith efficiently by performing grinding collectively is performed.

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

  However, conventionally, there has been a problem that it is difficult to position the embedding position when embedding the otolith which is the observation sample in a resin or the like. For this reason, when otoliths are fixed, the inclination and the fixed position are not constant, and it is difficult to form an observation cross section that can be observed entirely by the otolith nucleus and the ring around it. .

  The present invention has been made in view of the above-mentioned circumstances, and it is possible to easily and accurately form an observation cross section including a otolith nucleus of a fish, and a primary embedding block for observation of an otolith, an ear It is an object of the present invention to provide a method for producing a stone observation specimen block and an otolith sample piece.

That is, the primary embedding block for otolith observation of the present invention has the following configuration.
The base material has a flat sample fixing surface for fixing a fish otolith sample, and the sample fixing surface is formed with a positioning index indicating a position of a nucleus of the otolith sample.

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

  Further, the present invention is characterized in that the positioning index is a crossing line consisting of two straight lines crossing each other at the center point of the nucleus.

  Further, the present invention is characterized by further comprising a processing index line which is formed at a position separated from the central point and which indicates a distance to the central point along one straight line of the intersecting lines.

  Further, according to the present invention, an auxiliary index line having one end extending toward the central point and the one end spaced apart from the central point along a direction inclined at a predetermined angle with respect to the intersection line. Furthermore, it is characterized by having.

The otolith observation sample block of the present invention has the following configuration.
From the otolith observation primary embedding block, the otolith sample fixed to the sample fixing surface, and a transparent covering member covering the otolith observation primary embedding block to which the otolith sample is fixed It is characterized by becoming.

The method for producing an otolith sample piece according to the present invention has the following configuration.
A method for producing an otolith sample piece using the otolith observation sample block, wherein the otolith sample is fixed on the sample fixing surface such that a nucleus of the otolith sample coincides with the center point of the intersection line. An otolith fixation step of fixing the otolith sample, and a covering step of covering the primary embedding block for observation for otoliths to which the otolith sample is fixed with a transparent covering member, and forming the specimen block for observation for otoliths; And at least one polishing step of shaving the otolith observation sample block from two directions toward the nucleus of the otolith sample along one straight line forming the second The sample block for observation of otoliths is scraped to a position where a part of the nuclei of the otolith sample is exposed while grasping the distance to the nucleus of the otolith sample with reference to the index line.

  According to the otolith observation primary embedding block, the otolith observation specimen block, and the otolith sample piece production method of the present invention, it is possible to easily and accurately form an observation cross section including a fish otolith nucleus Becomes possible.

It is a perspective view and a side view showing a primary embedding block for otolith observation according to an embodiment of the present invention in a state where an otolith (otolith sample) is fixed. It is a perspective view and a side view showing a primary embedding block for otolith observation according to another embodiment of the present invention in a state where an otolith (otolith sample) is fixed. It is a top view which shows the crossing line formed in a sample fixed surface, an auxiliary index line, and an auxiliary index line. It is a perspective view showing the sample block for otolith observation of one embodiment of the present invention. It is explanatory drawing which shows the manufacture process of the sample block for otolith observation. It is a perspective view which shows an example of an otolith sample piece.

  Hereinafter, with reference to the drawings, a method for producing a primary embedding block for observation of otoliths, a specimen block for observation of otoliths, and a piece of otolith specimens according to an embodiment of the present invention will be described. Each embodiment shown below is concretely described in order to understand the meaning of the invention better, and does not limit the present invention unless otherwise specified. Further, in the drawings used in the following description, for the sake of easy understanding of the features of the present invention, the main parts may be enlarged for convenience, and the dimensional ratio of each component may be the same as the actual one. Not necessarily.

FIG. 1 (a) is a perspective view showing a primary embedding block for observation of otoliths according to the present invention in a state where an otolith (a otolith sample) is fixed. FIG. 1 (b) is a side view along the Y-axis direction of FIG. 1 (a). Moreover, FIG.1 (c) is a side view along the X-axis direction of Fig.1 (a).
The primary embedding block 10 for otolith observation includes a base material 11 having a flat sample fixing surface 11 a on which the otolith sample S is fixed. The base material 11 forms a rectangular parallelepiped in this embodiment, and an arbitrary surface is a sample fixing surface 11a. The base 11 is made of a material that is easy to polish, for example, a resin material. In addition, the resin material needs to be formed of a transparent (light transmitting) resin material.

On the sample fixing surface 11a, a crossing line (virtual crossing line) 13 formed of two straight lines 13a and 13b indicating a center point C indicating the position of the otolith nucleus SC located at the center of the otolith sample S is formed. . Such an intersecting line 13 is an index indicating the position of the axis (short axis, long axis) of the otolith sample S.
In the present embodiment, the intersecting 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, an axis parallel to the straight line 13a is referred to as an X axis, an axis parallel to the straight line 13b is referred to as a Y axis, and an axis perpendicular to the X axis and the Y axis is referred to as a Z axis. Such crossing lines 13 are made of, for example, ink, such as toner.

In the otolith sample S, the otolith sample S is placed on the sample fixation surface 11a such that the otolith nucleus SC at the center of the otolith sample S coincides with the center point C of the intersecting line 13 formed on the sample fixation surface 11a. It is fixed to The otolith nucleus SC can be easily recognized as a black dot, for example, by transmitting light to the otolith sample S. For this reason, the base material 11 uses a light transmitting resin.
The otolith sample S and the sample fixing surface 11a may be fixed using, for example, an ultraviolet curable resin. Thereby, the otolith sample S can be quickly fixed to the sample fixing surface 11a.

  In addition, on the sample fixing surface 11a, one of the two straight lines 13a and 13b, which is a processing index line 14a, 14b, 14c, 14d disposed at a predetermined distance along the Y axis in this embodiment Is formed. The processing index lines 14a to 14d indicate the separation width along the Y axis from the center point C of the intersection line 13. In the present embodiment, the processing index line 14a extends from the center point C of the intersection line 13 to the Y axis. Along the Y axis and the processing index line 14b is from the center point C of the intersection line 13 along the Y axis to the position of +1.0 mm, the center line C of the intersection line 13 from the center point C from the Y axis The processing index line 14d is formed at a position of -0.5 mm along the Y-axis from the center point C of the intersection line 13 at a position of -0.5 mm. The lengths of the processing index lines 14a to 14d themselves are formed to be 1.0 mm in the present embodiment. The processing index lines 14a to 14d are made of, for example, ink.

  Furthermore, in the sample fixing surface 11a, an auxiliary index whose one end extends toward the center point C and one end is at a distance from the center point C along a direction inclined at a predetermined angle with respect to the intersecting line 13. Lines 15a-15d are formed. In the present embodiment, the auxiliary index lines 15 a to 15 d extend at an angle of 45 ° with respect to the intersection line 13, for example. The length of the auxiliary index lines 15a to 15d themselves is formed to be 1.0 mm in the present embodiment. The auxiliary index lines 15a to 15d are made of, for example, ink.

  The intersecting lines 13 formed on the sample fixing surface 11a as described above, the processing index lines 14a to 14d, and the auxiliary index lines 15a to 15d are shown in FIG. As shown in FIG. 3, 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 central point C as an actual line. A circular area may be set as the central point C. The auxiliary index lines 15a to 15d are also imaginary lines near 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. In the embodiment shown in FIG. 1, the otolith sample S is fixed to the sample fixing surface 11a such that the major axis of the elliptical otolith sample S is along the straight line 13a of the intersecting line 13, ie, the X axis.

  As in the embodiment shown in FIGS. 2 (a) and 2 (b), the otolith sample is such that the minor axis of the elliptical otolith sample S is along the straight line 13b of the intersecting line 13, ie, the X axis. S can also be fixed to the sample fixing surface 11a.

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

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

Next, a method for producing otolith sample pieces using the otolith observation sample block 20 will be described with reference to FIGS. 1 and 5 including the action of the primary embedding block 10 for otolith observation.
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 according to the present invention, first, the primary for otolith observation according to the present invention Prepare embedded block 10. Then, with the central point C of the intersecting line 13 formed on the sample fixing surface 11 a of the primary embedding block 10 for otolith observation upward, a small amount of curable resin is formed in the vicinity thereof.

Next, the otolith sample S is placed on the sample fixing surface 11a, and light transmitted through the otolith sample S is irradiated,
For example, the otolith nucleus SC present in the otolith sample S 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 11 a so that the otolith nucleus SC coincides with the central point C of the intersection line 13.
Furthermore, the otolith sample S is rotated so that the major axis of the elliptical otolith sample S coincides with the straight line 13 a (X axis) or the straight line 13 b (Y axis) forming the crossing line 13.
Then, the hardenable resin is hardened, 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).

  Thus, the ear of the otolith sample S is formed by forming the intersecting line 13 consisting 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. A straight line 13a (X-axis) or a straight line 13b (Y-axis) in which the stone nucleus SC overlaps the center point C of the intersecting line 13 and the direction of the major axis of the elliptical otolith sample S forms the intersecting line 13 The otolith sample S can be fixed to the sample fixing surface 11 a so that it can be easily grasped that any of the above is matched.

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

  In addition, as a transparent resin material which comprises the covering member 21, the same thing as the resin material which comprises the base material 11 of the primary embedding block 10 for otolith observation is used. Moreover, as an example of the transparent resin material which comprises the covering member 21, ultraviolet-ray cured resin, a thermosetting resin, a photocurable resin etc. are mentioned.

  Next, the otolith observation sample block 20 is placed on the polishing apparatus, and from two directions toward the otolith nucleus SC of the otolith sample S, along the straight line 13b forming the intersecting line 13, ie, the Y axis, The stone observation sample block 20 is polished (polishing step).

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

  Then, when the surface to be polished comes to the positions of the processing index line 14a and the processing index line 14c, polishing is performed at a low polishing rate, and the otolith nucleus SC and its surroundings are removed without scraping off the otolith nucleus SC. The observation cross section of the entire surrounding ring can be easily and reliably formed.

In the polishing after the positions of the processing index line 14a and the processing index line 14c, the auxiliary index lines 15a to 15d, the straight lines 13a (X axis), and the straight lines 13b (Y By polishing while confirming the distance from the axis), the distance from the polished surface to the otolith nucleus SC can be accurately grasped by the trigonometric function, and the otolith nucleus SC is more reliably scraped off The observation cross section of the target of the otolith sample S can be formed without.
Also, when the otolith sample S is large, the sample block 20 for otolith observation is placed on the cutting apparatus 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 obtained in this manner is shown in FIG.
In the otolith sample piece 30 shown in FIG. 6, the otolith sample S is polished from both sides along the X-axis of the otolith observation specimen block 20 (see FIG. 4) to form a disc.

  While certain embodiments have been described, these embodiments have been presented by way of example only, and are not intended to limit the scope of the invention. These embodiments can be implemented in other various forms, and various omissions, replacements, and modifications can be made without departing from the scope of the invention. These embodiments and modifications thereof are included in the invention described in the claims and the equivalents thereof as well as included in the scope and the gist of the invention.

For example, in the above-described embodiment, a cross line is formed as a core positioning index, but in addition to this, for example, a line serving as a center point and a long axis or a short axis index, a center point and a long axis and a short axis A line serving as an index of the axis, a center point and a circular area can be used as a positioning index of the nucleus, and the specific form of the positioning index of the nucleus is not limited.
Moreover, although the shape of the primary embedding block for otolith observation is made into the rectangular parallelepiped in embodiment mentioned above, it can be made various shapes, such as hexagonal plate shape and disk shape besides this, for example. Moreover, a plurality of primary embedding blocks for observation of otoliths can be simultaneously formed using a mold capable of forming a plurality of primary embedding blocks for observation of otoliths. As a result, a large number of otolith samples can be collectively fixed to the primary embedding block for otolith observation, and this can be covered with a covering member to efficiently form observation cross sections of a large number of otolith samples become.
Moreover, in the embodiment described above, the otolith observation sample block 20 is formed in a cylindrical shape, but in addition to this, for example, various shapes such as a cube shape, a hexagonal column shape, etc. can be used.
In addition, the configuration of the primary embedding block for otolith observation according to the present invention can be applied, for example, to embed hard tissues (bones, shells, mollusk otoliths, pearls, etc.) of other organisms. In this case, it is possible to determine the direction of polishing and the position during polishing based on the intersection line, the processing index line, and the auxiliary index line as the observation target corresponding to the nucleus of each organism.

DESCRIPTION OF SYMBOLS 10 ... Primary embedding block for otolith observation 11 ... Base material 11a ... Sample fixed surface 13 ... Crossing line 13a, 13b ... Straight line 14a-14d ... Machining index line 15a-15d ... Auxiliary index line 20 ... Sample block for otolith observation 21: Coating member C: Center point S: Otolith sample (otolith)
SC: Otolith nucleus Q: Resin material liquid

Claims (6)

A substrate having a flat sample fixing surface for fixing fish otolith samples;
A primary embedding block for observation of otoliths, wherein a positioning index indicating a position of a nucleus of the otolith sample is formed on the sample fixing surface.   The primary embedding block for otolith observation according to claim 1, wherein the positioning index is a crossing line consisting of two straight lines crossing each other at the center point of the nucleus.   The otolith observation according to claim 2, further comprising a processing index line formed at a position separated from the center point and indicating a distance to the center point along one straight line of the intersecting lines. Primary embedding block.   It further comprises an auxiliary index line extending at one end toward the center point and at a distance from the center point along a direction inclined at a predetermined angle with respect to the intersection line. The primary embedding block for otolith observation according to claim 2 or 3 characterized by the above.   A transparent covering covering the primary embedding block for otolith observation according to claim 4, the otolith sample fixed on the sample fixing surface, and the primary embedding block for otolith observation on which the otolith sample is fixed. A specimen block for observation of otoliths, comprising: a member; A method for producing an otolith sample piece using the sample block for otolith observation according to claim 5,
An otolith fixation step of fixing the otolith sample to the sample fixing surface such that a nucleus of the otolith sample coincides with the center point of the intersection line;
Covering the otolith observation primary embedding block to which the otolith sample is fixed with a transparent covering member to form the otolith observation sample block;
Polishing at least abrading the otolith observation specimen block from two directions toward the nucleus of the otolith sample along one straight line forming the intersecting line;
In the polishing step, the sample for observing the otolith to a position where a part of the nucleus of the otolith sample is exposed while grasping the distance to the nucleus of the otolith sample with reference to the processing index line and the auxiliary index line The manufacturing method of the otolith sample piece characterized by scraping a block.

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