CN101539495B - Preparation of mackerel otolith cross-sectional slices and method for determining age thereof - Google Patents
Preparation of mackerel otolith cross-sectional slices and method for determining age thereof Download PDFInfo
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- CN101539495B CN101539495B CN2009100469831A CN200910046983A CN101539495B CN 101539495 B CN101539495 B CN 101539495B CN 2009100469831 A CN2009100469831 A CN 2009100469831A CN 200910046983 A CN200910046983 A CN 200910046983A CN 101539495 B CN101539495 B CN 101539495B
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Abstract
The invention relates to a preparation of mackerel otolith cross-sectional slices and a method for determining age thereof, the preparation of the cross-sectional slice comprises the following steps of: immersing, washing otolith, line-drawing, removing otolith, burning, embedding resin, incising, abrading, polishing and the like. Mackerel otolith slice is placed on a glass slide coated with dark blue coating, a few drops of glycerine is applied to cover the mackerel otolith slice below a CCD anatomical lens, interpretation is carried out on the rings under the irradiation of reflected light to determine the age of otolith. The invention realizes the clarity and visibility of winter ring and summer ring of mackerel otolith, avoids the interference of crystallization, solar disk and the summer ring on growth ring, enhances the color contrast between the winter ring and the summer ring, and facilitates the winter ring and the growth ring to be identified and interpreted, thus improving the precision for determining the age of mackerel; in addition, the slices of otolith can be preserved for a long time.
Description
Technical field
The present invention relates to a kind of making and age authentication method thereof of mackerel otolith cross-sectional slices, be specifically related to the method that a kind of mackerel otolith cross-sectional slices of making by the calcination method and age thereof identify, belong to the Fisheries Science field.
Background technology
Mackerel is one of important economic fish in China coastal waters, is the basis of carrying out the assessment of mackerel resources science to the reasonable estimation of mackerel growth parameter(s), and is a very important basic work to the accurate evaluation at mackerel age.
Fish are initial from hatching, and otolith just forms, and wheel of translucent winter and wheel of opaque summer all deposit on otolith every year afterwards, have formed annual ring jointly.Just can determine the age of fish according to the wheel line number on the otolith.Therefore, how, determine that its annual ring number is the key technical problem of identifying at the mackerel age exactly in addition interpretation of the wheel line on the mackerel otolith.
Scale, vertebrae, otolith all once were used to identify the age of mackerel.The mackerel scale is little and thin, be difficult for gathering and preserving, and the winter takes turns and the extremely difficult differentiation of false wheel, the age is identified brought very big difficulty.Though the annual ring on the mackerel I-V vertebrae is apparent in view, its three-dimensional conical in shape makes annual ring also be difficult to observe at microscopically, and the collection of vertebrae must cut the fish body, is cumbersome therefore.
It is to decide the most frequently used method of mackerel age that the otolith surface is observed, but the mackerel otolith is a milky and translucent, winter wheel and summer wheel are not easily distinguishable, and the otolith surface distributed has numerous wheel lines, make the sun, annual ring and summer wheel be difficult to distinguish, the age evaluation is caused very big difficulty, the age of the ground interpretation mackerel that leads to errors easily.When there was crystalline solid in the otolith surface, crystalline solid covered the wheel line, can't carry out the age to it and identify.Therefore, this method has been strengthened the color contrast of winter wheel and summer wheel by otolith is carried out calcination, and by the wheel line on the interpretation otolith xsect, overcome the interference of the sun and crystalline solid, year taken turns to identification, thereby reached the purpose that improves the evaluation accuracy of mackerel age but improved to annual ring.
Summary of the invention
Technical matters to be solved by this invention provides a kind of making and age authentication method thereof of mackerel otolith cross-sectional slices, but this method can effectively improve the identification and the accuracy at mackerel age.
In order to achieve the above object, the present invention realizes by the following technical solutions:
A kind of method for making of mackerel otolith cross-sectional slices may further comprise the steps:
1) soak: after the mackerel otolith is taken out, put into container, the adding weight percent concentration is 70% alcohol, soaks for 1 week;
2) clean otolith: the mackerel otolith is taken out from container, put into the double dish that fills water, under anatomical lens, clean up with the organic organization and the impurity of small brushes with mackerel otolith remained on surface;
3) setting-out: after the mackerel otolith cleans up, observe under anatomical lens, concave surface is found out the core of mackerel otolith up, then with mark line that passes mackerel otolith core of pencil drawing, draws a line of cut at position, mackerel otolith proparea;
4) excision otolith: under anatomical lens, with tweezers the mackerel otolith is fixed on the other hand, another hand, excises mackerel otolith proparea part along line of cut with scalper;
5) calcination: the mackerel otolith after will cutting takes out from double dish, is placed on the thieving paper water is blotted, and then the mackerel otolith is placed on the wave carrier piece that has concave surface, uses the spirit lamp calcination to brown or dark-brown;
6) resin embedding: by 1: 1.5 volume ratio acryl powder and hardening agent are configured to solution, pour solution into mould of plastics, solution covers mold bottom and gets final product, the otolith cut surface up and be parallel to mold bottom continues to add solution, till filling it up with, placed 24 hours in lucifuge, solidify;
7) cutting: the mould that will be embedded with the mackerel otolith is the center with the mark line, cuts into the thin slice of 0.8-1.2mm;
8) polishing: mackerel otolith section is placed on the grinding machine polishing dish, is that the waterproof abrasive paper of P1200 and P2000 is polished thick to 0.3-0.7mm with specification respectively;
9) polishing: it is standby that specification is that the burnishing powder of 0.3 μ m is configured to the suspending liquid of 70～90g/L, changes waterproof abrasive paper into polishing flannelet, sucks burnishing powder suspending liquid with syringe, evenly drips to be sprinkling upon on the flannelette polishing both surfaces of will cutting into slices then.
Described mackerel age authentication method is, microslide is simultaneously covered fully with mazarine coating, dries standby; The section of mackerel otolith is positioned on the microslide that scribbles mazarine coating, under the CCD anatomical lens, and drips last several glycerine, under the reflected light irradiation, the wheel line is carried out interpretation.
Used acryl powder and hardening agent among the present invention, chemical name is a methyl methacrylate, is the import resin material of metallographic style special use, is applicable to fields such as PCB, FPC, semiconductor technology, microelectric technique, medical technology, photoelectric technology.At room temperature acryl powder and hardening agent are mixed by proper proportion, can staticly solidify, generate high molecular polymer--transparent resin.It solidifies rapidly, solidification effect ideal (set time: 6min); Can not damage human body, nontoxic; Safe in utilization, easy.
Make mackerel cross-sectional slices of the present invention and age thereof when identifying, pay particular attention to following some:
(1) mackerel otolith thin and fragile can not obtain xsect by direct cutting.Therefore use resin embedding, obtain the xsect of mackerel otolith again by cutting, polishing.Xsect should pass through the otolith core, comprises complete annual ring information to guarantee xsect.
(2) calcination time and extent control.The mackerel otolith can not directly be used the flame calcination, should be placed on the microslide, uses spirit lamp flame envelope calcination 3-5 second, when the mackerel otolith becomes brown or brownish red by white, stops calcination immediately.When mackerel otolith color is faint yellow, illustrate that calcination time is not enough, can continue calcination to brown or brownish red.If the calcination of mackerel otolith is excessive, color changes yellow-gray into by brown, and the mackerel otolith is then opaque fully.
(3) the mackerel otolith is after calcination, under radiating light, and its cross-section center, the white ovals agglomerate of opacity is the core of mackerel otolith, and the peripheral transparent yellowish-brown band of mackerel otolith core is winter wheel or summer wheel, and the opaque and white band is the summer wheel.The summer wheel occurs through the winter wheel edge of being everlasting, and imports the winter wheel.Winter takes turns relatively, and the summer wheel is thinner, and imperfect, is the branch of winter wheel.
The present invention realized to mackerel otolith winter wheel and summer take turns clear visual, avoided crystallization, the sun, summer to take turns interference to annual ring, strengthened the color contrast of winter wheel and summer wheel, make winter wheel and more easy identification and interpretation of annual ring, thereby improved the accuracy of mackerel age evaluation, the otolith section also can be preserved for a long time.
Description of drawings
Further specify the present invention below in conjunction with the drawings and specific embodiments.
Fig. 1 is the structural representation of mackerel otolith profile of the present invention (concave surface) and mark line, line of cut.
Wherein 10 is the proparea, and 11 are the Qian Bei district, and 20 is the back zone, and 21 is back porch, and 30 are back of the body district, and 40 is the abdomen district, and 50 is line of cut, and 60 is core, and 70 is mark line.
Fig. 2 is the structural representation of winter wheel, summer wheel and mackerel otolith core on the mackerel otolith xsect of the present invention.
Embodiment
For technological means, creation characteristic that the present invention is realized, reach purpose and effect is easy to understand, below in conjunction with concrete diagram, further set forth the present invention.
The making of mackerel otolith cross-sectional slices:
1) soaks: after the mackerel otolith takes out, put into the centrifuge tube of the 1ml volume of underlined number, add 70% alcohol, soaked for 1 week, to remove the part organic organization of mackerel otolith remained on surface; And with waterproof marking pen mark.
2) clean otolith: the mackerel otolith is taken out, put into the double dish that fills water, under anatomical lens from centrifuge tube, clean up with the organic organization and the impurity of small brushes mackerel otolith remained on surface, when avoiding calcination, the contaminating impurity otolith that organic organization produces, influence is observed.
3) setting-out: after the mackerel otolith cleans up, under anatomical lens, observe, concave surface up, find out the core 60 of mackerel otolith, pass the mark line 70 of mackerel otolith core then with one of pencil drawing, draw a line of cut 50 at 10 positions, mackerel otolith proparea, line of cut 50 should leave the enough distances of mark line 70, damages mackerel otolith core 60 when avoiding cutting.
4) excision otolith: with reference to Fig. 1, under anatomical lens, with tweezers the mackerel otolith is fixed on the other hand, another hand, excises mackerel otolith proparea 10 parts along line of cut 50 with scalper.
5) calcination: the mackerel otolith after will cutting takes out from double dish, be placed on the thieving paper water is blotted, then the mackerel otolith is placed on the wave carrier piece that has concave surface, use spirit lamp calcination 3-5 second, when the otolith color is brown or dark-brown by leucismus, stop heating.Should evenly heat during calcination, keep a close eye on the otolith change color.Prevent the not enough or excessive calcination of otolith calcination, thereby influence the color contrast of winter wheel and summer wheel.Calcination is not enough, and color contrast is strong inadequately; Calcination is excessive, and winter wheel and summer wheel then become opaque fully, with the fubaritic age.
6) resin embedding: earlier with the mould of plastics label, number is consistent with mackerel otolith label.Mould is an opening, hollow rectangular parallelepiped: long 22.5mm, wide 9mm, high 10mm, wall thickness 1.5mm, base thickness 2mm.To be configured to solution by the acryl powder and the hardening agent of 1: 1.5 volume ratio, solution will be poured in the mould of plastics.Solution covers mold bottom and get final product, and mackerel otolith cut surface and be parallel to mold bottom continues adding solution, till filling it up with up.Placed 24 hours at the lucifuge place, make the solution completion of cure.When noting configuration solution, should avoid bubble to occur, bubble may appear on the mackerel otolith cross section as far as possible, the interpretation of influence wheel line.
7) cutting: the mould that will be embedded with the mackerel otolith is fixed on the cutting machine anchor clamps, is the center with the mark line 70 on mackerel otolith surface, is cut to the thin slice of 1mm on cutting machine.
8) polishing: the section of mackerel otolith is placed on the grinding machine polishing dish, is the center with the mark line, and it is thick to 0.5mm to polish with the waterproof abrasive paper of specification P1200 and P2000.
9) polishing: the suspending liquid of burnishing powder configuration 70～90g/L that with specification is 0.3 μ m is standby, changes waterproof abrasive paper into polishing flannelet, sucks burnishing powder suspending liquid with syringe, evenly drips to be sprinkling upon on the flannelette polishing both surfaces of will cutting into slices then.
Through above-mentioned 9 steps, mackerel otolith cross-sectional slices completes.
Embodiment 2
The evaluation at mackerel age:
With the even colored blue coating of microslide one side, dry adapted.
The section of mackerel otolith is positioned on the microslide, covers section, under blue background and reflected light irradiation, carry out annual ring interpretation and counting and take pictures enlargement factor 10～100 with the microscope of being with CCD with glycerine.Under reflected light, winter wheel 80, summer wheel 90 alternately exist.The winter wheel 80 yellowish-brown bands that are translucent, summer wheel 90 is the opaque and white band.Mackerel otolith core 60 is the opaque and white agglomerate, and tiny summer wheel 90 is arranged on every side.The summer wheel 90 yellowish-brown bands that also are translucent under reflected light follow the winter wheel to exist usually, note distinguishing winter wheel 80 and summer wheel 90.With respect to winter wheel 80, summer wheel 90 is thinner, and imperfect.
To winter wheel note row counting, winter wheel 80 numbers are the age.Each mackerel otolith is counted respectively all by two people interpretation alone.If it is identical that two people count, think that then the age interpretation is correct, accept this result; If the annual ring number difference of two people counting, then interpretation again, counting, and the result discussed.The back is discussed has identical ideas, then accept, otherwise abandon.
More than show and described ultimate principle of the present invention and principal character and advantage of the present invention.The technician of the industry should understand; the present invention is not restricted to the described embodiments; that describes in the foregoing description and the instructions just illustrates principle of the present invention; without departing from the spirit and scope of the present invention; the present invention also has various changes and modifications, and these changes and improvements all fall in the claimed scope of the invention.The claimed scope of the present invention is defined by appending claims and equivalent thereof.
Claims (2)
1. the method for making of a mackerel otolith cross-sectional slices is characterized in that, may further comprise the steps:
1) soak: the mackerel otolith is put into container after taking out, and the adding weight percent concentration is 70% alcohol, soaks for 1 week;
2) clean otolith: the mackerel otolith is taken out from container, put into the double dish that fills water, under anatomical lens, clean up with the organic organization and the impurity of small brushes with mackerel otolith remained on surface;
3) setting-out: after the mackerel otolith cleans up, observe under anatomical lens, concave surface is found out the core of mackerel otolith up, then with mark line that passes mackerel otolith core of pencil drawing, draws a line of cut at position, mackerel otolith proparea;
4) excision otolith: under anatomical lens, with tweezers the mackerel otolith is fixed on the other hand, another hand, excises mackerel otolith proparea part along line of cut with scalper;
5) calcination: the mackerel otolith after will cutting takes out from double dish, is placed on the thieving paper water is blotted, and then the mackerel otolith is placed on the wave carrier piece that has concave surface, uses the spirit lamp calcination to brown or dark-brown;
6) resin embedding: by 1: 1.5 volume ratio acryl powder and hardening agent are configured to solution, pour solution into mould of plastics, solution covers mold bottom and gets final product, mackerel otolith cut surface up and be parallel to mold bottom continues to add solution, till filling it up with, placed 24 hours at the lucifuge place, solidify;
7) cutting: the mould that will be embedded with the mackerel otolith is the center with the mark line, cuts into the thin slice of 0.8-1.2mm;
8) polishing: the section of mackerel otolith is placed on the grinding machine polishing dish, and it is thick to 0.3-0.7mm to polish with the waterproof abrasive paper of P1200 specification;
9) polishing: it is standby that specification is that the burnishing powder of 0.3 μ m is configured to the suspending liquid of 70～90g/L, changes waterproof abrasive paper into polishing flannelet, sucks burnishing powder suspending liquid with syringe, evenly drips to be sprinkling upon on the flannelette polishing both surfaces of will cutting into slices then.
2. mackerel age authentication method is characterized in that, mackerel otolith cross-sectional slices method for making according to claim 1 is made the section of mackerel otolith; Microslide is simultaneously covered fully with mazarine coating, dry standby; Mackerel otolith section is positioned over scribbles on the mazarine coating microslide, under the CCD anatomical lens, drip last several glycerine and cover the section of a mackerel otolith, under the reflected light irradiation, the wheel line is carried out interpretation.
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN102172882B (en) * | 2011-01-28 | 2013-01-30 | 上海海洋大学 | Method for grinding and preserving cephalopod otolith slices |
| CN102539212B (en) * | 2012-01-29 | 2013-08-21 | 山东理工大学 | Manufacture process for section of skin placoid scale of cartilaginous fish through adopting metallographic sample method |
| CN103231285B (en) * | 2013-04-26 | 2015-10-21 | 中国科学院水生生物研究所 | A kind of preparation method of mandarin fish otolith cross section abrasive disc and application thereof |
| CN103518655A (en) * | 2013-10-14 | 2014-01-22 | 中国水产科学研究院长江水产研究所 | Method for manufacturing section of xenophysogobio boulengeri otolith |
| CN103843708A (en) * | 2014-03-17 | 2014-06-11 | 上海海洋大学 | Method for constructing saury otolith physiological state analyzing model |
| CN105158035B (en) * | 2015-08-20 | 2017-12-15 | 江苏省海洋水产研究所 | The preparation method that a kind of little yellow croaker larva and juvenile otolith sagittal plane for micro-chemical analysis is cut into slices |
| CN106813968B (en) * | 2017-02-09 | 2018-10-12 | 中国水产科学研究院黄海水产研究所 | A kind of preparation method of hairtail sagitta cross section |
| CN112507533B (en) * | 2020-11-24 | 2023-03-14 | 上海海洋大学 | Method for constructing iris cuttlefish growth model based on marine environmental factors |
| CN114740003A (en) * | 2022-04-24 | 2022-07-12 | 烟台大学 | Method for rapidly identifying age of crustacean |
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| CA2008903A1 (en) * | 1989-03-09 | 1990-09-09 | Arun Narayan Netravali | Dual rail dilated switching networks |
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CA2008903A1 (en) * | 1989-03-09 | 1990-09-09 | Arun Narayan Netravali | Dual rail dilated switching networks |
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