CN106970079A - A kind of method that utilization inner casing identifies squids age in days - Google Patents
A kind of method that utilization inner casing identifies squids age in days Download PDFInfo
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- CN106970079A CN106970079A CN201710107533.3A CN201710107533A CN106970079A CN 106970079 A CN106970079 A CN 106970079A CN 201710107533 A CN201710107533 A CN 201710107533A CN 106970079 A CN106970079 A CN 106970079A
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
- G01N21/84—Systems specially adapted for particular applications
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N1/00—Sampling; Preparing specimens for investigation
- G01N1/28—Preparing specimens for investigation including physical details of (bio-)chemical methods covered elsewhere, e.g. G01N33/50, C12Q
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Abstract
The present invention relates to Fisheries Science field, the method that specifically a kind of utilization inner casing identifies squids age in days, grinding including inner casing cross section and the observation of growth line and the method counted, mainly comprise the following steps extractions, cleaning, dehydration, drying, cut out, resin embedding, cutting, fixation, polishing, polishing, picture making and grow the processes such as line counting.The present invention solves inner casing not easy grinding, and best fit plane can be quick and precisely ground to there is provided one kind by grinding the problem of easy to fall off and growth line is difficult to observe and counted, it is ensured that higher film-making success rate, while accurately interpretation grows the method for line number.
Description
Technical field
The present invention relates to Fisheries Science field, it is related to the observation procedure that a kind of inner casing grows line, is a kind of utilization specifically
The method that inner casing identifies squids age in days.
Background technology
Squids is Mollusca, Cephalopoda, gun shaped mesh, and the general name of squid section species, the life-span is about one year old, they
It is important marine economy species, is distributed widely in the marine sites such as each ocean in the world and south, the arctic.Age in days identification is used as squids fishing
The key content of industry Ecology, all the time by domestic and foreign scholars extensive concern.Otolith, Angle Overlap Model etc. are considered as
It is the reliable material for identifying squids age in days, the past also has research to attempt to identify squid using the growth line of inner housing surface
The age in days of class, but it is not used to age in days identification because the growth line interpretation of its early stage is difficult.
The content of the invention
It is an object of the invention to provide a kind of utilization inner casing identify squids age in days method, mainly comprise the following steps extraction,
Clean, be dehydrated, dry, cut out, resin embedding, cutting, fixation, polishing, polishing, picture making and growth line count etc. process；
The cross section of inner casing stem is polished, i.e., from inner casing proximally and distally respectively to grinding flakiness in the middle part of stem；Make
Good section is taken pictures with CCD, to ensure that growth line can be read at any time from now on；Line is grown by inner casing dorsal part core hemad side
Edge is counted.
This method is extracted using form stable, easily, grinds convenient inner casing to identify the age in days of squids, including squid
The making step of class inner casing section and observation and the counting step of inner casing growth line, by the growth line number for reading inner casing cross section
Mesh identifies the age in days of squids, for the age in days of Direct Identification squids provides a new method.
To achieve these goals, there is provided the side that a kind of utilization inner casing identifies squids age in days for the first aspect of the present invention
Method, comprises the following steps：
A, cut out：Take out inner casing, interception inner casing stem；
B, embedding：By 1:1~1:1.5 volume ratio by acryl powder and liquid curing-agent be configured to cold to bury resin molten
Liquid, is poured into mould, solution is wrapped up whole stem, and full of mould, then place and treat that it is hardened at room temperature lucifuge；
C, cutting：Cross-sectional cut resin thin slice of the resin along inner casing after hardening will be embedded；
D, fixation：The resin flake near-end that step c is obtained down, is fixed on slide；
E, polishing：The resin flake being fixed on slide distal end is polished, first roughly grinds and is finely ground to middle part again；
F, polishing：By the inner casing resin flake polishing grinding face through polishing, oil and impurity are removed, the resin is completed thin
The grinding of piece one side；
G, resin flake polished step f together with slide is placed on warm table heating until resin softens, removed
Resin flake, after drying, resin flake turn-over is fixed on slide, fine grinding again is then first roughly ground from distal end to middle part straight
Growth line to cross section is high-visible, and last re-polishing obtains inner casing section；
H, growth line observation：The number of growth line is counted by inner casing dorsal part core hemad edge.
It is preferred that, inner casing shank and tail bone portion are wiped out in the cutting in step a, leave behind stem.
It is preferred that, inner casing distal end can be also sticked on slide down, then be beaten from inner casing near-end to middle part in step d
Mill, polishing and implementation subsequent step.
It is preferred that, inner casing is pre-processed before being cut out in step a, comprised the following steps：
A, extraction：Squid inner casing is carefully extracted with scalpel, scissors, inner casing fracture is prevented, crushes；
B, cleaning：Inner housing surface is cleaned up with clear water；
C, dehydration：Inner casing after cleaning is put into absolute alcohol and is dehydrated 30 days；
D, drying：It will be put into through the step C inner casings handled in fume hood and allow it to spontaneously dry 1 week or so.
It is preferred that, the growth line counting step in step h is：
I, picture making：Section will be made to be placed on slide, supernatant water is dripped with glue head dropper；It is being connected with CCD's
Microscope 200 × times under the growth line of inner casing is taken pictures, picture mosaic is carried out respectively to the picture clapped with image processing software
Processing；
II, growth line are counted：200 × times lower the image gathered is counted by inner casing dorsal part core hemad edge and grown
The wider place of selection wheel spacing is counted in line number, counting process.
The second aspect of the present invention comprises the following steps there is provided the preparation method that a kind of squids inner casing is cut into slices：
A, cut out：Take out inner casing, interception inner casing stem；
B, embedding：By 1:1~1:1.5 volume ratio by acryl powder and liquid curing-agent be configured to cold to bury resin molten
Liquid, is poured into mould, solution is wrapped up whole stem, and full of mould, then place and treat that it is hardened at room temperature lucifuge；
C, cutting：Cross-sectional cut resin thin slice of the resin along inner casing after hardening will be embedded；
D, fixation：The resin flake near-end that step c is obtained down, is fixed on slide；
E, polishing：By the resin flake near-end being fixed on slide polishing, first roughly grind and be finely ground to middle part again；
F, polishing：By the inner casing resin flake polishing grinding face through polishing, oil and impurity are removed, the resin is completed thin
The grinding of piece one side；
G, resin flake polished step f together with slide is placed on warm table heating until resin softens, removed
Resin flake, after drying, resin flake turn-over is fixed on slide, fine grinding again is then first roughly ground from distal end to middle part straight
Growth line to cross section is high-visible, and last re-polishing obtains inner casing section.
It is preferred that, inner casing is pre-processed before being cut out in step a, comprised the following steps：
A, extraction：Squid inner casing is carefully extracted with scalpel, scissors, inner casing fracture is prevented, crushes；
B, cleaning：Inner housing surface is cleaned up with clear water；
C, dehydration：Inner casing after cleaning is put into absolute alcohol and is dehydrated 30 days；
D, drying：It will be put into through the step C inner casings handled in fume hood and allow it to spontaneously dry 1 week or so.
It is furthermore preferred that the preparation method of above-mentioned squids inner casing section, comprises the following steps：
(1) extract：Squid inner casing is carefully extracted with scalpel, scissors, inner casing fracture is prevented, crushes；
(2) clean：Inner housing surface is cleaned up with clear water；
(3) it is dehydrated：Inner casing after cleaning is put into absolute alcohol and is dehydrated 30 days；
(4) dry：The inner casing handled through step (3) is put into fume hood and allows it to spontaneously dry 1 week or so.
(5) cut：Dried inner casing is taken out, shank and tail bone portion is wiped out, leaves stem standby；
(6) embed：By 1:1~1:1.5 volume ratio by acryl powder and liquid curing-agent be configured to cold to bury resin molten
Liquid, is poured into mould, solution is wrapped up whole stem, and full of mould, then place and treat that it is hardened at room temperature lucifuge；
(7) cut：Cross-sectional cut of the resin along inner casing after hardening will be embedded into 2-3mm resin flakes, and make grinding
Cut surface of the face parallel to inner casing；
(8) it is fixed：The resin flake (inner casing near-end is down) is bonded on slide with the fast viscose glue of strength；
(9) polish：By the resin flake being fixed on slide (inner casing distal end) polishing, in process of lapping, successively with
240th, 600,1200,2500 silicon carbide papers are ground to middle part；
(10) polish：Added water with 0.05 μm of aluminum oxide agent in flannelette polishing grinding face, remove the surface that silicon carbide paper is caused
Cut and unnecessary impurity, so complete the grinding of inner casing distal end；
(11) resin flake polished by more than is placed on heating on warm table until resin softening, is removed together with slide
Resin flake, after drying, resin flake turn-over is fixed on slide, then repeats above-mentioned from inner casing inlet side to middle part
Grinding and polishing process are until the growth line of cross section is high-visible, and process of lapping will constantly observe grinding under an optical microscope
Degree, in order to avoid overgrinding.
There is provided a kind of observation of squids inner casing growth line and method of counting, including following step for the third aspect of the present invention
Suddenly：
I, picture making：The inner casing made in above-mentioned steps section is placed on slide, supernatant water is dripped with glue head dropper；
Be connected with CCD microscope 200 × times under the growth line of inner casing is taken pictures, with image processing software to the picture clapped
Picture mosaic processing is carried out respectively；
II, growth line are counted：200 × times lower the image gathered is counted by inner casing dorsal part core hemad edge and grown
The wider place of selection wheel spacing is counted in line number, counting process.
The invention has the advantages that：
1st, the method that the utilization inner casing that the present invention is provided identifies squids age in days, inner casing easily extracts, grinds convenient, information
Stable storing, inner casing identification squids age in days is directly, reliably, accurately.
2nd, the inventive method, which chooses inner casing stem, is used for microsection manufacture, the standard for improving the readability of growth line and counting
True property；The grinding of inner casing buries resin embedding using cold in the inventive method, then cutting and grinding again, it is to avoid in inner casing process of lapping
The skew of middle ground flat, adds the success rate of microsection manufacture and the readability of growth line.
3rd, the inventive method is counted in growth line, and the convenience that growth line is counted is greatly facilitated using camera system；
The section made is taken pictures with CCD, to ensure that growth line can be read at any time from now on.
4th, the present invention solve inner casing be difficult directly grinding (added after embedding grinding contact surface, reduce ground flat
Deflection probability), grind (dried inner casing is difficult for drop-off after resin embedding) easy to fall off and growth line is difficult to observe and counted
The problem of number (inner casing stem cross section growth line is easy to observe and counted than other positions), can be quick and precisely there is provided one kind
Best fit plane is ground to, it is ensured that higher film-making success rate, while accurately interpretation grows the method for line number.
Brief description of the drawings
Shell morphology, orientation and cross-sectional view in Fig. 1 squidses.
Fig. 2 squidses inner casing grows line counting direction.
Embodiment
The embodiment provided with reference to embodiment the present invention elaborates.
Embodiment 1：The method that squids age in days is identified using inner casing
Operating procedure is as follows：
1st, extract：Squid inner casing is carefully extracted with scalpel, scissors, inner casing fracture is prevented, crushes；
2nd, clean：Inner casing after taking-up cleans the mucus and other organic matters on its surface with clear water；
3rd, it is dehydrated：Inner casing after cleaning is put into absolute alcohol and is dehydrated 30 days；
4th, dry：Inner casing after dehydration is put into fume hood and allows it to spontaneously dry 1 week or so.
5th, cut：Thoroughly dried inner casing (Fig. 1) is taken out, shank and tail bone portion is wiped out with surgical scissors, retains stem
Portion；
6th, embed：By 1:1~1:1.5 volume ratio by acryl powder and liquid curing-agent (viscosity is 60mPas, purchase in
The vast and boundless experimental instruments and equipment limited in Shenzhen three) be configured to it is cold bury resin solution, pour into mould, solution is wrapped up whole inner casing
Stem, and full of mould, then place and treat that it is hardened at room temperature lucifuge；
7th, cut：Cross-sectional cut of the resin along inner casing after hardening will be embedded into 2-3mm resin flakes, and make abradant surface
Parallel to the cut surface of inner casing；
8th, it is fixed：The resin flake (inner casing near-end is down) will be bonded on slide with the fast viscose glue of strength；
9th, polish：By the resin flake being fixed on slide (inner casing distal end) polishing, in process of lapping, successively with 240,
600th, 1200,2500 silicon carbide papers are ground to middle part；
10th, polish：Added water with 0.05 μm of aluminum oxide agent in flannelette polishing grinding face, remove the surface stroke that silicon carbide paper is caused
Trace and unnecessary impurity, so complete the grinding of inner casing distal end；
11st, resin flake polished by more than is placed on heating on warm table until resin softening, is removed together with slide
Resin flake, after drying, resin flake turn-over is fixed on slide, then repeats above-mentioned from inner casing inlet side to middle part
Grinding and polishing process are until the growth line of cross section is high-visible, and process of lapping will constantly observe grinding under an optical microscope
Degree, in order to avoid overgrinding；
12nd, the inner casing polished section is put into paper bag and preserved, and carry out mark；
13rd, picture making：Section will be made to be placed on slide, with the upper drop clear water of glue head dropper drop, (increase is observed
Definition)；Be connected with CCD (ccd image sensor) Olympus microscope 200 × times under to the growth line of inner casing
Taken pictures；With the image processing softwares of Photoshop 7.0, picture mosaic processing is carried out respectively to the picture clapped；
14th, growth line is counted：200 × times lower the image gathered is counted by inner casing dorsal part core hemad edge and grown
The wider place of selection wheel spacing is counted (Fig. 2) in line number (Fig. 2), counting process.
Embodiment 2：A kind of preparation method of squids inner casing section
Operating procedure is as follows：
(1) cut：Thoroughly dried inner casing (Fig. 1) is taken out, shank and tail bone portion is wiped out with surgical scissors, retains stem
Portion；
(2) embed：By 1:1~1:By acryl powder and liquid curing-agent, (viscosity is 60mPas to 1.5 volume ratio, buying
In the vast and boundless experimental instruments and equipment limited in Shenzhen three) be configured to it is cold bury resin solution, pour into mould, make solution parcel it is whole in
Shell stem, and full of mould, then place and treat that it is hardened at room temperature lucifuge；
(3) cut：Cross-sectional cut of the resin along inner casing after hardening will be embedded into 2-3mm resin flakes, and make grinding
Cut surface of the face parallel to inner casing；
(4) it is fixed：The resin flake (inner casing near-end is down) will be bonded on slide with the fast viscose glue of strength；
(5) polish：By the resin flake being fixed on slide (inner casing distal end) polishing, in process of lapping, successively with
240th, 600,1200,2500 silicon carbide papers are ground to middle part；Also inner casing distal end can be sticked on slide down, then by inner casing
Subsequent step is polished, polishes and implemented to middle part in distal end；
(6) polish：Added water with 0.05 μm of aluminum oxide agent in flannelette polishing grinding face, remove the surface stroke that silicon carbide paper is caused
Trace and unnecessary impurity, so complete the grinding of inner casing distal end；
(7) resin flake polished by more than is placed on heating on warm table until resin softening, is removed together with slide
Resin flake, after drying, resin flake turn-over is fixed on slide, then repeats above-mentioned from inner casing inlet side to middle part
Grinding and polishing process are until the growth line of cross section is high-visible, and process of lapping will constantly observe grinding under an optical microscope
Degree, in order to avoid overgrinding；
Inner casing is pre-processed, comprised the following steps before step (1) is cut out：
A, extraction：Squid inner casing is carefully extracted with scalpel, scissors, inner casing fracture is prevented, crushes；
B, cleaning：Inner housing surface is cleaned up with clear water；
C, dehydration：Inner casing after cleaning is put into absolute alcohol and is dehydrated 30 days；
D, drying：It will be put into through the step C inner casings handled in fume hood and allow it to spontaneously dry 1 week or so.
Embodiment 3：A kind of observation of squids inner casing growth line and method of counting
Operating procedure is as follows：
(1) picture making：Section will be made to be placed on slide, with the upper drop clear water of glue head dropper drop, (increase is observed
Definition)；Be connected with CCD (ccd image sensor) Olympus microscope 200 × times under to the growth line of inner casing
Taken pictures；With the image processing softwares of Photoshop 7.0, picture mosaic processing is carried out respectively to the picture clapped；
(2) growth line is counted：200 × times lower the image gathered is counted by inner casing dorsal part core hemad edge and grown
The wider place of selection wheel spacing is counted (Fig. 2) in line number (Fig. 2), counting process.
The preferred embodiment to the invention is illustrated above, but the invention be not limited to it is described
Embodiment, those skilled in the art can also make a variety of equivalent on the premise of without prejudice to the invention spirit
Modification or replacement, these equivalent modifications or replacement are all contained in the application claim limited range.
Claims (8)
1. a kind of method that utilization inner casing identifies squids age in days, it is characterised in that comprise the following steps：
A, cut out：Take out inner casing, interception inner casing stem；
B, embedding：By 1:1~1:1.5 volume ratio by acryl powder and liquid curing-agent be configured to it is cold bury resin solution, fall
Enter in mould, solution is wrapped up whole stem, and full of mould, then place and treat that it is hardened at room temperature lucifuge；
C, cutting：Cross-sectional cut resin thin slice of the resin along inner casing after hardening will be embedded；
D, fixation：The resin flake near-end that step c is obtained down, is fixed on slide；
E, polishing：The resin flake being fixed on slide distal end is polished, first roughly grinds and is finely ground to middle part again；
F, polishing：By the inner casing resin flake polishing grinding face through polishing, oil and impurity are removed, the resin flake one is completed
The grinding in face；
G, resin flake polished step f together with slide is placed on warm table heating until resin softens, removes resin
Thin slice, after drying, resin flake turn-over is fixed on slide, and fine grinding again is then first roughly ground from distal end to middle part until horizontal
The growth line in section is high-visible, and last re-polishing obtains inner casing section；
H, growth line observation：The number of growth line is counted by inner casing dorsal part core hemad edge.
2. the method that utilization inner casing according to claim 1 identifies squids age in days, it is characterised in that described step a
In cutting inner casing shank and tail bone portion are wiped out, leave behind stem.
3. the method that utilization inner casing according to claim 1 identifies squids age in days, it is characterised in that described step a
In cut out before inner casing is pre-processed, comprise the following steps：
A, extraction：Squid inner casing is carefully extracted with scalpel, scissors, inner casing fracture is prevented, crushes；
B, cleaning：Inner housing surface is cleaned up with clear water；
C, dehydration：Inner casing after cleaning is put into absolute alcohol and is dehydrated 30 days；
D, drying：It will be put into through the step C inner casings handled in fume hood and allow it to spontaneously dry 1 week or so.
4. the method that utilization inner casing according to claim 1 identifies squids age in days, it is characterised in that described step d
In also can by inner casing distal end stick at down on slide, subsequent step is then polished, polished and implemented to middle part from inner casing near-end.
5. the method that utilization inner casing according to claim 1 identifies squids age in days, it is characterised in that described step h
In growth line counting step be：
I, picture making：The inner casing made section is placed on slide, supernatant water is dripped with glue head dropper；It is being connected with CCD
Microscope 200 × times under the growth line of inner casing is taken pictures, the picture clapped is spelled respectively with image processing software
Figure processing；
II, growth line are counted：Growth line number is counted by inner casing dorsal part core hemad edge to the 200 × times lower image gathered
The wider place of selection wheel spacing is counted in mesh, counting process.
6. a kind of preparation method of squids inner casing section, it is characterised in that comprise the following steps：
A, cut out：Take out inner casing, interception inner casing stem；
B, embedding：By 1:1~1:1.5 volume ratio by acryl powder and liquid curing-agent be configured to it is cold bury resin solution, fall
Enter in mould, solution is wrapped up whole stem, and full of mould, then place and treat that it is hardened at room temperature lucifuge；
C, cutting：Cross-sectional cut resin thin slice of the resin along inner casing after hardening will be embedded；
D, fixation：The resin flake near-end that step c is obtained down, is fixed on slide；
E, polishing：By the resin flake near-end being fixed on slide polishing, first roughly grind and be finely ground to middle part again；
F, polishing：By the inner casing resin flake polishing grinding face through polishing, oil and impurity are removed, the resin flake one is completed
The grinding in face；
G, resin flake polished step f together with slide is placed on warm table heating until resin softens, removes resin
Thin slice, after drying, resin flake turn-over is fixed on slide, and fine grinding again is then first roughly ground from distal end to middle part until horizontal
The growth line in section is high-visible, and last re-polishing obtains inner casing section.
7. the preparation method of squids inner casing section according to claim 6, it is characterised in that cut in described step a
Inner casing is pre-processed before sanction, comprised the following steps：
A, extraction：Squid inner casing is carefully extracted with scalpel, scissors, inner casing fracture is prevented, crushes；
B, cleaning：Inner housing surface is cleaned up with clear water；
C, dehydration：Inner casing after cleaning is put into absolute alcohol and is dehydrated 30 days；
D, drying：It will be put into through the step C inner casings handled in fume hood and allow it to spontaneously dry 1 week or so.
8. observation and the method for counting of a kind of squids inner casing growth line, it is characterised in that comprise the following steps：
I, picture making：The section of squids inner casing is made using the preparation method described in claim 6 or 7, in making
Shell section is placed on slide, and supernatant water is dripped with glue head dropper；Be connected with CCD microscope 200 × times under life to inner casing
Long line is taken pictures, and picture mosaic processing is carried out respectively to the picture clapped with image processing software；
II, growth line are counted：Growth line number is counted by inner casing dorsal part core hemad edge to the 200 × times lower image gathered
The wider place of selection wheel spacing is counted in mesh, counting process.
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| Application Number | Priority Date | Filing Date | Title |
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| CN201710107533.3A CN106970079B (en) | 2017-02-27 | 2017-02-27 | Method for identifying day age of teleost by using inner shell |
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| CN201710107533.3A CN106970079B (en) | 2017-02-27 | 2017-02-27 | Method for identifying day age of teleost by using inner shell |
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| CN110243640A (en) * | 2018-03-08 | 2019-09-17 | 中国科学院海洋研究所 | The inner cavity of deep-sea foraminifer is ground fast preparation method |
| CN111693527A (en) * | 2020-06-05 | 2020-09-22 | 上海海洋大学 | Method for identifying day age of cuttlefish by using cuttlefish bone |
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Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110243640A (en) * | 2018-03-08 | 2019-09-17 | 中国科学院海洋研究所 | The inner cavity of deep-sea foraminifer is ground fast preparation method |
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