CN105738175B - A kind of fish early life history otolith sample preparation methods - Google Patents
A kind of fish early life history otolith sample preparation methods Download PDFInfo
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- CN105738175B CN105738175B CN201610125432.4A CN201610125432A CN105738175B CN 105738175 B CN105738175 B CN 105738175B CN 201610125432 A CN201610125432 A CN 201610125432A CN 105738175 B CN105738175 B CN 105738175B
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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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- 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
- G01N1/32—Polishing; Etching
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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
- G01N1/34—Purifying; Cleaning
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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
- G01N1/36—Embedding or analogous mounting of samples
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Abstract
The present invention relates to biological sample preparation fields, and in particular to a kind of fish early life history otolith sample preparation methods, this method is：Corrosive solution is added dropwise under mirror to expose otolith；The otolith that exposure is cleaned with purified water is multiple, while absorbing the liquid after cleaning and/or tissue remnants；Otolith is embedded with mountant after otolith drying at room temperature, is dried after mountant is paved；It also needs to carry out grinding processing before fish early life history middle and later periods otolith mounting.Method provided by the invention reduces the difficulty of fish early life history stage otolith acquisition, even if sample tissue is opaque can also to use this method, and sample obtained is cleaner, and impurity is less, is more advantageous to the long-term preservation of sample.
Description
Technical field
The present invention relates to biological sample preparation fields, in particular to a kind of fish early life history otolith sample system
Preparation Method.
Background technology
Ecology of fishes research includes mainly age, growth, feeding habits, death and Population fluctuation etc..Due to fish
Population fluctuation most depend on the early life history stage (embryonic period, embryonic phase, larva and juvenile phase and young stage) survival rate,
So the ecological study of early life history has ten for the supplement, population structure and population scale of biological species adult population
Divide great meaning.
With annalistic information, and continuously grown in entire life process, the fish bone with firm anatomical structure
Bone structure such as fish scale, nucleus, fin ray, vertebra etc. are generally used for the reference at fish age and upgrowth situation judgement.Os osseum
The structure of calcification at first is typically otolith in fish, and Fish otolith is located in inner ear, generally by sagitta, micro- otolith and star otolith
Each 1 pair of composition.Compared to other skeletal structures, the particularity of otolith, which is to grow, has continuity, is affected by the external environment opposite
Small, the fish of acquisition gained always do not find that otolith is reuptaked phenomenon in the wild, and just remain unchanged once being formed, because
And the good carrier permanently recorded as life historical event part.
Since Pannella (1971) describes the deposition growth variation of Fish otolith day for the first time, Fish otolith is studied
Extensive development is arrived, with the development of correlation theory and technology, it has been known as a kind of announcement fish history of life, the history of life rebuilds
And the key technology that water body environment domain becomes.The fish relevant ecological research in early life history stage is sent out in Otolith Morphology at present
Educate, the sun wheel identification age in days and the sun wheel increase etc. have more report.Otolith microstructure detection is mainly used for differentiating with analytical technology
Prelarva population speculates egg-laying time, prelarva early development condition and the assessment death rate, and it is raw to have become researching fish early stage at present
The important means of history living.Therefore, it is the important ring of Otolith microstructure to be tested and analyzed to prepare otolith sample.However, existing
Otolith preparation method mostly be the adult fish being directed to, it is directly covered in the preparation of larva and juvenile otolith, exist win otolith be stranded
Difficult problem.In addition that there is also samples is unholiness, the problem of being not easy to preserve for a long time for the prior art.
In view of this, special propose the present invention.
Invention content
The purpose of the present invention is to provide a kind of fish early life history otolith sample preparation methods, the preparation methods
Operation is easy to grasp, and can solve the fish in early life history and win that otolith is difficult, sample obtained is unholiness, it is permanent to be not easy
The problem of preservation.
In order to realize that the above-mentioned purpose of the present invention, spy use following technical scheme：
A kind of fish early life history otolith sample preparation methods, include the following steps：
1), the fish sample in early life history is placed under anatomical lens or stereomicroscope, on the fish sample
Corrosive solution is added dropwise to expose otolith；The corrosive solution is the solution of weak acid, weak base or strong base-weak acid salt；
2) otolith that exposure, is cleaned with purified water is multiple, while absorbing the liquid after cleaning and/or tissue remnants；
3), otolith is embedded with mountant after otolith drying at room temperature, is dried after mountant is paved.
The fish early life history stage as an important link in the fish entire history of life, early development stage at
Motility rate is directly related to the size of the practical magnitude of recruitment of fish, is the main original for causing Population fluctuation and age composition to change
Cause, and it is closely related using scale and protection of fish resources policy making etc. with fisheries resources exploitation.
Under normal circumstances, the fish early life history stage includes mainly embryo's (ovum), alevin stage, postlarva phase and young stage.
This method by stages is divided according to the development of fish organ structure, nutrition and the features such as ingest.
Fish otolith in the early life history stage is smaller, and sample volume also very little itself, thus otolith win it is non-
Often difficult, the application erodes the tissue around otolith by using corrosive solution so that otolith wins being easy to for change.
In addition, the application uses conventional pressed disc method, but the i.e. observable after drying mountant.It is made in this way
Sample it is cleaner, impurity is less, is more advantageous to the long-term preservation of sample, and suitable for the fish sample in period after body devitrification
Product.
The number that the otolith of exposure is cleaned with purified water is generally 3~4 times.
Preferably, corrosive solution is being added dropwise with sudden and violent in fish early life history otolith sample preparation methods as described above
In the step of revealing otolith, specifically include：
Dropwise addition corrosive solution is added dropwise purified water and slows down the speed that fish sample is corroded again.
Due to the diversity and complexity of biological sample, the composition etc. of the outer tissue thickness of different fish, tissue all may be used
Can difference, in order to enable the solution rate of fish body becomes controllable, so optionally, needing to dilute corrosivity with purified water
The concentration of liquid.
Preferably, the step of fish early life history otolith sample preparation methods as described above, the embedding, specifically wraps
It includes：
Mountant is dropped near otolith sample, organic solvent is added dropwise in the mountant makes mountant free diffusing
Otolith is embedded.
This investment fashion, mountant using organic solvent as carrier, under the action of surface tension slowly diffusion be paved with it is whole
A glass slide avoids the generation of bubble.The effect that organic solvent may also function as transparent otolith, dilute mountant, makes it be unlikely to
It is too sticky.
Preferably, sample preparation methods as described above, the fish sample include in embryonic period, embryonic phase, alevin stage, postlarva phase
Or fresh sample, Cord blood sample or the fixed sample of absolute ethyl alcohol of young stage.
Wherein embryonic period, embryonic phase is specially the embryonic period, embryonic phase after otolith has been formed.
Preferably, fish early life history otolith sample preparation methods as described above, when the fish sample is embryonic period, embryonic phase
Or when alevin stage sample, in step 1)~2) in, the exposure of otolith and cleaning method are specially：
When corrosive solution by the fish body corrosion around otolith it is clean, transparent when begin to wash with water, and leave residual
The fish body deposited is as the reference of otolith position.
On the one hand do so is since otolith is too small, kick the beam, easily slide, it is easy to which, in cleaning, filter paper is siphoned away；
On the other hand, for embryonic period, embryonic phase or alevin stage sample, since otolith is too small, there is no position object of reference, in microscope
It is difficult to find otolith, thus need before fish body is not completely dissolved when lower observation analysis, according to the fish body of remaining as ear
The reference of stone position not wait fish bodies to be completely dissolved only remaining otolith and clean again.
Preferably, fish early life history otolith sample preparation methods as described above, when the fish sample is the postlarva phase
Or when young stage sample, step 1) includes：
The fish head of the fish sample is cut, fish body is abandoned；
Corrosive solution is added dropwise in the fish head of the fish sample to submerge fish head, by ear at statocyst after fish head is soft
Stone is chosen and removes extra tissue.
The fish body of usual postlarva phase and young stage is just bigger, in order to easy to operate, should individually remove fish head.
The fish body tissue of postlarva phase is also more thick and solid, so after being softened sample with corrosive solution, after otolith is chosen.
Further include grinding before mounting after embedding it is further preferred that for otolith sample after 30 ages in days；
The operation of the grinding specifically includes：It is sudden and violent to its core with the embedded otolith of 2000~3000 mesh liquid honings
Dew, then polished the core exposed with the waterproof abrasive paper of 5000~10000 mesh, it is cleaned 2~3 times with purified water, uses organic solvent
Mountant is dissolved, the side that do not polish is placed and dried outwardly.
Otolith before usual preceding 30 age in days need not polish, and the otolith after 30 ages in days has had certain thickness,
It can not distinguish wheel line, need to carry out sanding and polishing.
Preferably, sample preparation methods as described above, the corrosive solution are liquor natrii hypochloritis.
Sodium hypochlorite is a kind of strong base-weak acid salt, and solution has corrosivity.
It is further preferred that when the fish sample is embryonic period, embryonic phase or alevin stage sample：
When the fish sample be embryonic period, embryonic phase or alevin stage sample when, the sodium hypochlorite be 5.5~6.5% sodium hypochlorite
Solution；
When the fish sample be postlarva phase or young stage sample when, the sodium hypochlorite be 10~13% sodium hypochlorite
Solution；
The sodium hypochlorite concentration is in terms of effective chlorine.
Preferably, fish early life history otolith sample preparation methods as described above：
The mountant is neutral gum, and the organic solvent is dimethylbenzene.
Neutral gum, as a kind of adhesive, for glass slide and coverslip to be bonded together, biological tissue section
It closes to retain for a long time.It has the advantages that following：
Transparency is high, almost clear as optical glass；
Not etching glass, because it is neutral, and glass is most afraid of alkalinity, if worried can use salt before bonding
Acid or alcohol immersion dry again glue may be more preferable；
It is not easy mildew, it may be possible to because the solvent xylene of neutral gum has killing fungus effect.
Xylene solubles or dilution neutral gum, and have the function of clarifier.
Compared with prior art, beneficial effects of the present invention are：
1) very small (19.8 μ of such as 3 age in days Pseudopleuronectes sagitta diameter of seawater fish otolith, in the early life history stage
M, micro- otolith diameter are only 13.3 μm), it is difficult to which otolith is chosen under anatomical lens.The application is dissolved by using corrosive solution
Fish body greatly reduces the difficulty for obtaining otolith.
2) if the sample size, acquired is excessive, it usually needs need first to preserve fish body Cord blood or absolute ethyl alcohol.Fish
Tissue degeneratiaon after body preserves, it is no longer transparent, with conventional method face more challenges when otolith is won, but this method is not
There are problems that this.
3), though pressed disc method is simple and fast, that there are otolith samples is unholiness, is not easy long-term preservation and is not suitable for
The fish sample in period after opaque or body devitrification.The application uses conventional pressed disc method, but by mountant
Drying.Sample obtained in this way is cleaner, and impurity is less, is more advantageous to the long-term preservation of sample, and not suitable for fish body
Transparent sample.
Description of the drawings
In order to more clearly explain the embodiment of the invention or the technical proposal in the existing technology, to embodiment or will show below
There is attached drawing needed in technology description to be briefly described.
Fig. 1 is the otolith of Pseudopleuronectes prelarva in embodiment 2；A：Micro- otolith；B：Sagitta；
Fig. 2 is the Fugu rubripes prelarva in digestion process in embodiment 3；A：Micro- otolith；B：Sagitta；
Fig. 3 is resolution in embodiment 3, Fugu rubripes prelarva otolith after cleaning；A：Micro- otolith；B：Sagitta；
Fig. 4 is the otolith in 62 age in days Pseudopleuronectes otolith acquisition process in embodiment 5；A：Micro- otolith；B：Sagitta；C：Star
Otolith；
Fig. 5 is 62 age in days Pseudopleuronectes sagittas not being ground after mounting in embodiment 5；
Fig. 6 is the 62 micro- otoliths of age in days Pseudopleuronectes not being ground after mounting in embodiment 5；
Fig. 7 is 62 age in days Pseudopleuronectes star otoliths not being ground after mounting in embodiment 5；
Fig. 8 is the 62 age in days Pseudopleuronectes sagittas prepared in embodiment 5；
Fig. 9 is the 62 micro- otoliths of age in days Pseudopleuronectes prepared in embodiment 5.
Specific implementation mode
Embodiment of the present invention is described in detail below in conjunction with embodiment, but those skilled in the art will
Understand, the following example is merely to illustrate the present invention, and is not construed as limiting the scope of the invention.It is not specified in embodiment specific
Condition person carries out according to conventional conditions or manufacturer's recommended conditions.Reagents or instruments used without specified manufacturer is
The conventional products that can be obtained by commercially available purchase.
Embodiment 1
A kind of fish early life history otolith sample preparation methods, include the following steps：
S11, the fish sample in early life history is placed under anatomical lens or stereomicroscope, on the fish sample
Corrosive solution is added dropwise to expose otolith；The corrosive solution is the solution of weak acid, weak base or strong base-weak acid salt；
S12, the otolith that exposure is cleaned with purified water are multiple, while absorbing the liquid after cleaning and/or tissue remnants；
S13, with mountant otolith is embedded after otolith drying at room temperature, is dried after mountant is paved.
Embodiment 2
A kind of transparent embryos phase or alevin stage otolith sample preparation methods, include the following steps：
S21, fresh Pseudopleuronectes prelarva is placed on to glass slide center, be placed under anatomical lens or stereomicroscope, adjusting is put
Big multiple is to the visual field appropriate.
S22, otolith obtain and cleaning
5.5% liquor natrii hypochloritis is added dropwise in fish body, allows fish body dipping in the solution, it, can if histolysis is too fast
Addition purified water suitably dilutes, to dissolve Fish tissue with appropriate, controllable solution rate；
When otolith is high-visible, is cleaned 3~4 times with purified water, absorb surrounding liquid with filter paper every time.It need to wait for that solution is molten
It begins to be cleaned with purified water when clean, transparent around solution otolith, leaves the fish body of remaining as the reference of otolith position, not wait
Fish body is completely dissolved only remaining otolith and cleans again.The otolith drying at room temperature that will be obtained.
S23, otolith mounting
After glass slide drying, a drop neutral gum is first added dropwise near otolith, then drips upper two on neutral gum again
Toluene, such neutral gum are spread using dimethylbenzene as carrier to periphery, fish body of the embedding with otolith.Then by embedded ear
Stone mounting is placed dries in 50~60 DEG C of baking ovens, you can observes simultaneously persistence.Obtained otolith sample is as shown in Figure 1.
Embodiment 3
A kind of opaque embryonic period, embryonic phase or alevin stage otolith sample preparation methods, include the following steps：
S31, the fresh sample of Fugu rubripes prelarva is placed on to glass slide center, is placed in anatomical lens or stereomicroscope
Under, amplification factor is adjusted to the visual field appropriate.
S32, otolith obtain and cleaning
A concentration of 6.5% liquor natrii hypochloritis is added dropwise in fish body, fish body dipping is allowed in the solution, if histolysis
Soon, purified water can be added suitably to dilute, to dissolve Fish tissue with appropriate, controllable solution rate；In digestion process
Fugu rubripes prelarva is as shown in Figure 2；
When otolith is high-visible, is cleaned 3~4 times with purified water, absorb surrounding liquid with filter paper every time.It need to wait for that solution is molten
It begins to be cleaned with purified water when clean, transparent around solution otolith, leaves the fish body of remaining as the reference of otolith position, not wait
Fish body is completely dissolved only remaining otolith and cleans again.Otolith is picked out with tapering dissecting needle, is placed near remaining fish body, room
Temperature is dry.
S33, otolith mounting
After glass slide drying, a drop neutral gum is first added dropwise near otolith, then drips upper dimethylbenzene on natural gum again,
Natural gum is spread using dimethylbenzene as carrier to periphery in this way, the ear that fish body or remaining fish body and picking of the embedding with otolith come out
Stone.Then embedded otolith mounting is placed and is dried in 50-60 DEG C of baking oven, you can observation and persistence.Preceding 30 age in days
Otolith by dimethylbenzene it is transparent after do not have to polishing can observation analysis.Obtained otolith is as shown in Figure 3.
Embodiment 4
A kind of postlarva phase or young stage otolith sample preparation methods, include the following steps：
S41, it the fixed Pseudopleuronectes fish head of absolute ethyl alcohol is sheared off with dissecting scissors is placed on glass slide center, be placed in anatomical lens
Or under stereomicroscope, amplification factor is adjusted to the visual field appropriate.
S42, otolith obtain and cleaning：Ammonium hydroxide is added dropwise in fish head, submerges entire fish head, is dipped to that fish head is soft to choose i.e.
It can.Then fish head rear portion is pressed with tweezers or dissecting needle, dissecting needle is gradually chosen along the approximate location of statocyst with another
Fish head tissue is taken, after otolith is exposed, otolith is moved into glass slide and is totally located, is removed remaining broken rotten tissue with filter paper.
Last otolith 3~4 times wash with distilled water, absorb surrounding liquid, drying at room temperature with filter paper every time.
S43, otolith mounting：After glass slide drying, neutral gum is added dropwise near otolith, then again on natural gum in drop
Dimethylbenzene, natural gum are spread using dimethylbenzene as carrier to periphery, and there is otolith in embedding residence.Then embedded otolith mounting is placed
It is dried in 50~60 DEG C of baking ovens, you can persistence.
Embodiment 5
A kind of postlarva phase or young stage otolith sample preparation methods, include the following steps：
S51, it the fish head of the Pseudopleuronectes of age in days on the 62nd of Cord blood is sheared off with dissecting scissors is placed on glass slide center, set
Under anatomical lens or stereomicroscope, amplification factor is adjusted to the visual field appropriate.
S52, otolith obtain and cleaning
13% liquor natrii hypochloritis is added dropwise in fish head, submerges entire fish head, is dipped to that fish head is soft to choose；
Fish head rear portion is pressed with tweezers or dissecting needle, dissecting needle is gradually chosen along the approximate location of statocyst with another
Fish head tissue is taken, after otolith is exposed, otolith is moved into glass slide and is totally located, is removed remaining broken rotten tissue with filter paper；
Otolith in scale removal process is as shown in Figure 4；
Otolith is cleaned with purified water 3~4 times, absorbs surrounding liquid with filter paper every time, residual chlorine after preventing glass slide from drying
Change sodium crystallization, drying at room temperature.
S53, otolith mounting
After glass slide drying, neutral gum is added dropwise near otolith, then drips upper dimethylbenzene on natural gum again, natural gum with
Dimethylbenzene is that carrier is spread to periphery, and there is otolith in embedding residence.Then embedded otolith mounting is placed into 50~60 DEG C of baking ovens
In dry.
S54, grinding and fixation
Otolith after 30 ages in days has had certain thickness, can not distinguish wheel line, needs to carry out sanding and polishing.Prelarva, postlarva
Early period, otolith was without the use of sand paper sanding and polishing relative to relatively thin at fish otolith.
1) it, is polished embedded otolith with 2000~3000 mesh waterproof abrasive papers, it is micro- in low power after often polishing several seconds
Under the microscope, until being polishing at otolith core.
2), it is polished with the waterproof abrasive paper of 5000~10000 mesh again, is finally cleaned 2~3 times with purified water, with wiping
Mirror cloth blots surrounding.
3) melted paraxylene, is added to dissolve neutral gum, overturns otolith, making not polish, that is faced outwardly, baking oven drying.
It according to step 1), 2) operates, final obtain can be with the Pseudopleuronectes fish juvenile fish otolith sample of observation analysis and persistence
Product, these samples are as shown in Fig. 5~9.
Although illustrate and describing the present invention with specific embodiment, it will be appreciated that without departing substantially from the present invention's
Many other change and modification can be made in the case of spirit and scope.It is, therefore, intended that in the following claims
Including belonging to all such changes and modifications in the scope of the invention.
Claims (10)
1. a kind of fish early life history otolith sample preparation methods, which is characterized in that include the following steps：
1), the fish sample in early life history is placed under anatomical lens or stereomicroscope, is added dropwise on the fish sample
Corrosive solution is to expose otolith；The corrosive solution is the solution of weak acid, weak base or strong base-weak acid salt；
2) otolith that exposure, is cleaned with purified water is multiple, while absorbing the liquid after cleaning and/or tissue remnants；
3), otolith is embedded with mountant after otolith drying at room temperature, is dried after mountant is paved.
2. fish early life history otolith sample preparation methods as described in claim 1, which is characterized in that corrosivity is being added dropwise
In the step of solution is to expose otolith, specifically include：
Dropwise addition corrosive solution is added dropwise purified water and slows down the speed that fish sample is corroded again.
3. fish early life history otolith sample preparation methods as described in claim 1, which is characterized in that the step of the embedding
Suddenly it specifically includes：
Mountant is dropped near otolith sample, organic solvent is added dropwise in the mountant makes mountant free diffusing by ear
Stone embeds.
4. fish early life history otolith sample preparation methods as described in claim 1, which is characterized in that the fish sample packet
Include fresh sample, Cord blood sample or the fixed sample of absolute ethyl alcohol in embryonic period, embryonic phase, alevin stage, postlarva phase or young stage
This.
5. fish early life history otolith sample preparation methods as claimed in claim 4, which is characterized in that when the fish sample
For embryonic period, embryonic phase or alevin stage sample when, in step 1)~2) in, the exposure of otolith and cleaning method are specially：
When corrosive solution by around otolith fish body corrosion it is clean, transparent when begin to wash with water, and leave remaining
Fish body is as the reference of otolith position.
6. fish early life history otolith sample preparation methods as claimed in claim 4, which is characterized in that when the fish sample
For postlarva phase or young stage sample when, step 1) includes：
The fish head of the fish sample is cut, fish body is abandoned；
Corrosive solution is added dropwise in the fish head of the fish sample to submerge fish head, is chosen otolith at statocyst after fish head is soft
Go out and removes extra tissue.
7. fish early life history otolith sample preparation methods as claimed in claim 6, which is characterized in that after 30 ages in days
Otolith sample further includes grinding before mounting after embedding；
The operation of the grinding specifically includes：It is exposed with the embedded otolith of 2000~3000 mesh liquid honings to its core,
The core exposed is polished with the waterproof abrasive paper of 5000~10000 mesh again, is cleaned 2~3 times with purified water, will be sealed with organic solvent
The side that do not polish is placed and is dried outwardly by tablet dissolved.
8. the fish early life history otolith sample preparation methods as described in claim 1,2,5 or 6, which is characterized in that described
Corrosive solution is liquor natrii hypochloritis.
9. fish early life history otolith sample preparation methods as claimed in claim 8, it is characterised in that：
When the fish sample is embryonic period, embryonic phase or alevin stage sample, the sodium hypochlorite that the sodium hypochlorite is 5.5~6.5% is molten
Liquid；
When the fish sample be postlarva phase or young stage sample when, the sodium hypochlorite be 10~13% liquor natrii hypochloritis；
The sodium hypochlorite concentration is in terms of effective chlorine.
10. the fish early life history otolith sample preparation methods as described in claim 3 or 7, it is characterised in that：
The mountant is neutral gum, and the organic solvent is dimethylbenzene.
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| CN111829810B (en) * | 2020-07-20 | 2021-06-29 | 中国科学院水生生物研究所 | Method for obtaining fish otolith by ultrasonic separation |
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| CN105136541A (en) * | 2015-09-21 | 2015-12-09 | 中国水产科学研究院淡水渔业研究中心 | Pretreatment method for micro-chemical analysis of otoliths of Coilia nasus |
| CN105158035A (en) * | 2015-08-20 | 2015-12-16 | 江苏省海洋水产研究所 | Production method of little yellow croaker larval otolith sagittal plane sections for micro-chemical analysis |
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2016
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Patent Citations (8)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2004004449A1 (en) * | 2002-07-04 | 2004-01-15 | Veterinærmedisinsk Oppdragssenter As | Use of one or more chemical compound for marking of fish |
| JP2008067648A (en) * | 2006-09-14 | 2008-03-27 | Fisheries Research Agency | Agent and method for labeling fish |
| CN203259356U (en) * | 2013-05-17 | 2013-10-30 | 中国水产科学研究院南海水产研究所 | Negative pressure type otolith collector for small fishes |
| 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 |
| CN104198260A (en) * | 2014-09-15 | 2014-12-10 | 中国水产科学研究院黑龙江水产研究所 | Method for acquiring clean long-placed fish otolith |
| CN105158035A (en) * | 2015-08-20 | 2015-12-16 | 江苏省海洋水产研究所 | Production method of little yellow croaker larval otolith sagittal plane sections for micro-chemical analysis |
| CN105136541A (en) * | 2015-09-21 | 2015-12-09 | 中国水产科学研究院淡水渔业研究中心 | Pretreatment method for micro-chemical analysis of otoliths of Coilia nasus |
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