CN105806678A - Preparation method of material for identifying age of blue shark - Google Patents
Preparation method of material for identifying age of blue shark Download PDFInfo
- Publication number
- CN105806678A CN105806678A CN201610316002.0A CN201610316002A CN105806678A CN 105806678 A CN105806678 A CN 105806678A CN 201610316002 A CN201610316002 A CN 201610316002A CN 105806678 A CN105806678 A CN 105806678A
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- vertebrae
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- vertebral body
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- 239000000463 material Substances 0.000 title claims abstract description 30
- 241000251125 Prionace glauca Species 0.000 title claims abstract description 29
- 238000002360 preparation method Methods 0.000 title claims abstract description 18
- 238000000227 grinding Methods 0.000 claims abstract description 12
- 238000005498 polishing Methods 0.000 claims abstract description 8
- HEMHJVSKTPXQMS-UHFFFAOYSA-M Sodium hydroxide Chemical compound [OH-].[Na+] HEMHJVSKTPXQMS-UHFFFAOYSA-M 0.000 claims description 18
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 18
- 244000137852 Petrea volubilis Species 0.000 claims description 9
- 239000012153 distilled water Substances 0.000 claims description 9
- 238000009835 boiling Methods 0.000 claims description 6
- 210000002808 connective tissue Anatomy 0.000 claims description 6
- 238000007654 immersion Methods 0.000 claims description 6
- 230000001681 protective effect Effects 0.000 claims description 6
- UFMZWBIQTDUYBN-UHFFFAOYSA-N cobalt dinitrate Chemical compound [Co+2].[O-][N+]([O-])=O.[O-][N+]([O-])=O UFMZWBIQTDUYBN-UHFFFAOYSA-N 0.000 claims description 4
- 229910001981 cobalt nitrate Inorganic materials 0.000 claims description 4
- 238000004043 dyeing Methods 0.000 claims description 4
- 229920000297 Rayon Polymers 0.000 claims description 3
- 230000005540 biological transmission Effects 0.000 claims description 3
- 238000001035 drying Methods 0.000 claims description 3
- 239000003292 glue Substances 0.000 claims description 3
- 238000000034 method Methods 0.000 abstract description 9
- 230000008569 process Effects 0.000 abstract description 4
- 238000010719 annulation reaction Methods 0.000 abstract 2
- 238000004140 cleaning Methods 0.000 abstract 1
- 238000000605 extraction Methods 0.000 abstract 1
- 238000007789 sealing Methods 0.000 abstract 1
- 238000007447 staining method Methods 0.000 abstract 1
- 208000031481 Pathologic Constriction Diseases 0.000 description 6
- 210000001215 vagina Anatomy 0.000 description 6
- 241000251468 Actinopterygii Species 0.000 description 5
- 239000011295 pitch Substances 0.000 description 4
- 241000251730 Chondrichthyes Species 0.000 description 3
- 230000000694 effects Effects 0.000 description 3
- 238000010586 diagram Methods 0.000 description 2
- 238000012797 qualification Methods 0.000 description 2
- 238000012360 testing method Methods 0.000 description 2
- 230000009286 beneficial effect Effects 0.000 description 1
- 238000009395 breeding Methods 0.000 description 1
- 230000001488 breeding effect Effects 0.000 description 1
- 210000000845 cartilage Anatomy 0.000 description 1
- 238000004040 coloring Methods 0.000 description 1
- 238000005516 engineering process Methods 0.000 description 1
- 238000004519 manufacturing process Methods 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 238000011084 recovery Methods 0.000 description 1
- 238000011160 research Methods 0.000 description 1
- 230000001568 sexual effect Effects 0.000 description 1
- 238000010186 staining Methods 0.000 description 1
- 230000007704 transition Effects 0.000 description 1
Classifications
-
- 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
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N1/00—Sampling; Preparing specimens for investigation
- G01N1/02—Devices for withdrawing samples
- G01N1/04—Devices for withdrawing samples in the solid state, e.g. by cutting
- G01N1/06—Devices for withdrawing samples in the solid state, e.g. by cutting providing a thin slice, e.g. microtome
-
- 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
- G01N1/2813—Producing thin layers of samples on a substrate, e.g. smearing, spinning-on
-
- 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
- G01N1/286—Preparing specimens for investigation including physical details of (bio-)chemical methods covered elsewhere, e.g. G01N33/50, C12Q involving mechanical work, e.g. chopping, disintegrating, compacting, homogenising
-
- 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
- G01N1/30—Staining; Impregnating ; Fixation; Dehydration; Multistep processes for preparing samples of tissue, cell or nucleic acid material and the like for analysis
-
- 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
- G01N1/286—Preparing specimens for investigation including physical details of (bio-)chemical methods covered elsewhere, e.g. G01N33/50, C12Q involving mechanical work, e.g. chopping, disintegrating, compacting, homogenising
- G01N2001/2866—Grinding or homogeneising
-
- 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
- G01N1/286—Preparing specimens for investigation including physical details of (bio-)chemical methods covered elsewhere, e.g. G01N33/50, C12Q involving mechanical work, e.g. chopping, disintegrating, compacting, homogenising
- G01N2001/2873—Cutting or cleaving
Landscapes
- Life Sciences & Earth Sciences (AREA)
- Health & Medical Sciences (AREA)
- General Health & Medical Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Analytical Chemistry (AREA)
- Biochemistry (AREA)
- Physics & Mathematics (AREA)
- General Physics & Mathematics (AREA)
- Immunology (AREA)
- Pathology (AREA)
- Engineering & Computer Science (AREA)
- Biomedical Technology (AREA)
- Molecular Biology (AREA)
- Investigating Or Analysing Biological Materials (AREA)
- Farming Of Fish And Shellfish (AREA)
Abstract
The invention discloses a preparation method of a material for identifying the age of a blue shark. Different materials are used by vertebrae of different sizes, a big vertebra material is prepared by virtue of a longitudinal slicing method, and a small vertebra material is prepared by virtue of a staining method. According to the preparation method, the problems that the vertebrae of the blue shark is difficult to grind and is easily damaged during the grinding, and the definition of annulations of slices is low are solved; the grinding of longitudinal slices of the blue shark comprises the processes of extraction, cleaning, slicing, fixing, polishing and the like; and the longitudinal sections of the vertebrae of the blue shark are polished with uniform force, and the polished slices of the vertebrae are sealed by virtue of a sealing agent, so that the annulations can be maintained for a long time.
Description
Technical field
The preparation method that the present invention relates to a kind of marine organisms Age Determination material, particularly to the preparation method of a kind of blue shark Age Determination material.
Background technology
Blue shark is the important kind in cartilaginous fish, is positioned at the top of marine ecosystems, and as the main fishing kind of holding concurrently of longline fishery, its economic worth is higher.Vertebrae has the feature of storage-stable growth information, is research shark class age and the maximally effective material of growth；Vertebrae is double concave type, and on vertebrae, annual ring is characterized as that broad opaque layer (wide layer) and narrow semitransparent layer (narrow layer) are alternately arranged, in concentric circles.Wide layer is formed at temperature and rises growth rapid spring and summer season, and narrow layer is formed at temperature degrowth autumn and winter season slowly, and wide layer and the narrow layer intersection in next year then are annual ring；Blue shark vertebrae as a kind of growth information carrier, the growth information storing body input that it can be stable, discloses the information such as growth of young baby's length of blue shark, age of fish body and Ge Nian, therefore becomes the optimal material of shark class Age Determination.
Blue shark has poor growth, period of sexual maturity evening, the feature such as breeding cycle length, life-span length, once fished for by transition, the recovery difficulty of Stock resoures, therefore it is essential that blue shark carries out stock assessment, and age and growth information are the essential informations of assessment Stock resoures state；The vertebrae of blue shark grows along with the growth of fish body, but the continued growth along with fish body, vertebrae there will be growth arrest phenomenon, but the growth information of fish body still may proceed to be stored in vertebrae, so at shark Later growth, wheel stricture of vagina overlapping phenomenon easily occurs, therefore, select which kind of Age Determination method most important.
Summary of the invention
The preparation method that the technical problem to be solved in the present invention is to provide a kind of blue shark Age Determination material.
For reaching above-mentioned purpose, technical scheme is as follows:
The preparation method of a kind of blue shark Age Determination material, it is characterised in that different size vertebraes uses different materials, and big individual vertebrae material preparation method comprises the steps:
(1) extract: take about 3-5 from first dorsal place and save vertebrae, freezen protective；
(2) clean: vertebrae is placed in boiling water and boils about 5min, remove unnecessary connective tissue, if still unclean, it is placed in 0.5% sodium hydroxide solution and soaks, size according to vertebrae, soak time is about 8h-12h, after as about 30min in distilled water immersion；
(3) section: vertebrae is fixed on low speed microtome, and longitudinal section is cut to 1-2mm, by the nearly vertebral body median plane of longitudinal section 1200#Sand paper on be polishing to vertebral body plane, on buffing machine polish abradant surface；
Fixing: being bonded on microscope slide with powerful speed viscose glue by longitudinal section, vertebral body median plane is down；
Abrasive disc: with 800#Sand paper, 1200#Sand paper fine grinding section, and dissect its degree of grinding of Microscopic observation at transmission light at any time, to close to vertebral body center, section grinding thickness is 0.4-0.5mm, polishing abradant surface on buffing machine；
Mounting: microscope slide bubble is taken off section after 5-10 minute in 60 DEG C of water, after drying with the fixing vertebrae longitudinal section of PermountTM mountant on microscope slide, covered；
The preparation method of little individual vertebrae material comprises the steps of:
(1) extract: take about 3-5 from first dorsal place and save vertebrae, freezen protective；
(2) clean: vertebrae is placed in boiling water and boils about 5min, remove unnecessary connective tissue, if still unclean, it is placed in 0.5% sodium hydroxide solution and soaks, the size according to vertebrae, soak time is about 8h-12h, after as about 30min in distilled water immersion, and dry；
(3) dyeing: dry complete vertebrae is placed in 24h in 10% cobalt nitrate solution, after taking-up, and distilled water wash clean, finally dry.
In one embodiment of the invention, the blue shark that individual greatly vertebrae is fork length >=220cm is individual, and described little individual vertebrae is the blue shark individuality pitching long < 220cm.
By technique scheme, the invention has the beneficial effects as follows:
The inventive method is that big individual (fork length >=220cm) and the blue shark Age Determination material of little individuality (pitching long < 220cm) are prepared, find that the long individual blue shark little of 220cm of fork is suitable for utilizing complete vertebrae staining to carry out material and prepares according to testing, determine that cobalt nitrate dyeing is for the best by testing；And pitch up to relatively big to 220cm and the corresponding vertebrae of above sample, wheel stricture of vagina there will be overlap at edge, selects to carry out sections observation, and owing to having part cartilage in vertebrae material, longitudinal section prepares material effect can be more preferably.
The film-making of blue shark vertebrae is a complexity and fine process, and the quality of production effect is directly connected to accuracy and the reliability of Age Determination；Stain processes the definition that can deepen annual ring, and the accuracy identified is particularly important.
The present invention makes to take turns stricture of vagina on complete vertebrae by colouring method and becomes apparent from, solve vertebrae cut into slices not easy grinding, the problem such as easily damaged in process of lapping, section wheel stricture of vagina definition is not high, there is provided a kind of can the preparation method of quick and precisely different size blue shark Age Determination material, the material prepared shortens qualification time greatly, ensure higher qualification success rate, make the vertebrae after polishing take turns the time that stricture of vagina can keep longer simultaneously.
Accompanying drawing explanation
In order to be illustrated more clearly that the embodiment of the present invention or technical scheme of the prior art, the accompanying drawing used required in embodiment or description of the prior art will be briefly described below, apparently, accompanying drawing in the following describes is only some embodiments of the present invention, for those of ordinary skill in the art, under the premise not paying creative work, it is also possible to obtain other accompanying drawing according to these accompanying drawings.
Fig. 1 is the blue shark vertebrae form schematic diagram that the present invention pitches length >=220cm；
Fig. 2 is the blue shark vertebrae form longitudinal section schematic diagram that the present invention pitches length >=220cm；
Fig. 3 is the blue shark vertebrae colored graph that the present invention pitches long < 220cm；
Numeral and the corresponding component title represented by letter in figure:
10, vertebral body 20, vertebral body center 30, vertebral body median plane 40, growth ring stricture of vagina.
Detailed description of the invention
For the technological means making the present invention realize, creation characteristic, reach purpose and effect and be easy to understand, below in conjunction with being specifically illustrating, the present invention is expanded on further.
The preparation method that the invention discloses a kind of blue shark Age Determination material, different size vertebraes uses different materials；
Referring to shown in Fig. 1 and Fig. 2, big individual vertebrae (fork length >=220cm) material preparation method comprises the steps:
(1) extract: take about 3-5 from first dorsal place and save vertebrae, freezen protective；
(2) clean: vertebrae is placed in boiling water and boils about 5min, remove unnecessary connective tissue, if still unclean, it is placed in 0.5% sodium hydroxide solution and soaks, actual size according to vertebrae, soak time is about 8h-12h, after as about 30min in distilled water immersion；
(3) section: vertebrae is fixed on low speed microtome, and longitudinal section is cut to 1-2mm, by the nearly vertebral body median plane of longitudinal section 1200#Sand paper on be polishing to vertebral body plane, on buffing machine polish abradant surface；
Fixing: being bonded on microscope slide with powerful speed viscose glue by longitudinal section, vertebral body median plane is down；
Abrasive disc: with 800#Sand paper, 1200#Sand paper fine grinding section, and dissect its degree of grinding of Microscopic observation at transmission light at any time, to close to vertebral body center, section grinding thickness is 0.4-0.5mm, polishing abradant surface on buffing machine；
Mounting: microscope slide bubble is taken off section after 5-10 minute in 60 DEG C of water, after drying with the fixing vertebrae longitudinal section of PermountTM mountant on microscope slide, covered；
Shown in Figure 3, the preparation method of little individual vertebrae (pitching long < 220cm) material comprises the steps of:
(1) extract: take about 3-5 from first dorsal place and save vertebrae, freezen protective；
(2) clean: vertebrae is placed in boiling water and boils about 5min, remove unnecessary connective tissue, if still unclean, it is placed in 0.5% sodium hydroxide solution and soaks, the actual size according to vertebrae, soak time is about 8h-12h, after as about 30min in distilled water immersion, and dry；
(3) dyeing: dry complete vertebrae is placed in 24h in 10% cobalt nitrate solution, after taking-up, and distilled water wash clean, finally dry.
The ultimate principle of the present invention and principal character and advantages of the present invention have more than been shown and described.Skilled person will appreciate that of the industry; the present invention is not restricted to the described embodiments; described in above-described embodiment and description is that principles of the invention is described; 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 both fall within the claimed scope of the invention.Claimed scope is defined by appending claims and equivalent thereof.
Claims (2)
1. the preparation method of a blue shark Age Determination material, it is characterised in that different size vertebraes uses different materials, and big individual vertebrae material preparation method comprises the steps:
(1) extract: take about 3-5 from first dorsal place and save vertebrae, freezen protective；
(2) clean: vertebrae is placed in boiling water and boils about 5min, remove unnecessary connective tissue, if still unclean, it is placed in 0.5% sodium hydroxide solution and soaks, size according to vertebrae, soak time is about 8h-12h, after as about 30min in distilled water immersion；
(3) section: vertebrae is fixed on low speed microtome, and longitudinal section is cut to 1-2mm, by the nearly vertebral body median plane of longitudinal section 1200#Sand paper on be polishing to vertebral body plane, on buffing machine polish abradant surface；
Fixing: being bonded on microscope slide with powerful speed viscose glue by longitudinal section, vertebral body median plane is down；
Abrasive disc: with 800#Sand paper, 1200#Sand paper fine grinding section, and dissect its degree of grinding of Microscopic observation at transmission light at any time, to close to vertebral body center, section grinding thickness is 0.4-0.5mm, polishing abradant surface on buffing machine；
Mounting: microscope slide bubble is taken off section after 5-10 minute in 60 DEG C of water, after drying with the fixing vertebrae longitudinal section of PermountTM mountant on microscope slide, covered；
The preparation method of little individual vertebrae material comprises the steps of:
(1) extract: take about 3-5 from first dorsal place and save vertebrae, freezen protective；
(2) clean: vertebrae is placed in boiling water and boils about 5min, remove unnecessary connective tissue, if still unclean, it is placed in 0.5% sodium hydroxide solution and soaks, the size according to vertebrae, soak time is about 8h-12h, after as about 30min in distilled water immersion, and dry；
(3) dyeing: dry complete vertebrae is placed in 24h in 10% cobalt nitrate solution, after taking-up, and distilled water wash clean, finally dry.
2. the preparation method of a kind of blue shark Age Determination material according to claim 1, it is characterized in that, the blue shark that individual greatly vertebrae is fork length >=220cm is individual, and described little individual vertebrae is the blue shark individuality pitching long < 220cm.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201610316002.0A CN105806678B (en) | 2016-05-12 | 2016-05-12 | A kind of preparation method of blue shark Age Determination material |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201610316002.0A CN105806678B (en) | 2016-05-12 | 2016-05-12 | A kind of preparation method of blue shark Age Determination material |
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| Publication Number | Publication Date |
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| CN105806678A true CN105806678A (en) | 2016-07-27 |
| CN105806678B CN105806678B (en) | 2019-04-23 |
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN112729982A (en) * | 2020-12-21 | 2021-04-30 | 上海海洋大学 | Preparation method of material for identifying age of shark |
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Non-Patent Citations (10)
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN112729982A (en) * | 2020-12-21 | 2021-04-30 | 上海海洋大学 | Preparation method of material for identifying age of shark |
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