CN108651337A - A kind of production Drifting egg fish early stage different developmental phases form microscopy observation method - Google Patents
A kind of production Drifting egg fish early stage different developmental phases form microscopy observation method Download PDFInfo
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- CN108651337A CN108651337A CN201810460288.9A CN201810460288A CN108651337A CN 108651337 A CN108651337 A CN 108651337A CN 201810460288 A CN201810460288 A CN 201810460288A CN 108651337 A CN108651337 A CN 108651337A
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- 238000000034 method Methods 0.000 title claims abstract description 59
- 241000251468 Actinopterygii Species 0.000 title claims abstract description 22
- 238000004519 manufacturing process Methods 0.000 title claims abstract description 18
- 238000000386 microscopy Methods 0.000 title claims abstract description 15
- 239000012528 membrane Substances 0.000 claims abstract description 44
- 210000001161 mammalian embryo Anatomy 0.000 claims abstract description 43
- 210000003205 muscle Anatomy 0.000 claims abstract description 18
- 230000000694 effects Effects 0.000 claims abstract description 17
- 230000004720 fertilization Effects 0.000 claims abstract description 10
- 230000000877 morphologic effect Effects 0.000 claims abstract description 9
- 230000001418 larval effect Effects 0.000 claims abstract description 7
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims description 31
- 239000007788 liquid Substances 0.000 claims description 24
- 235000019441 ethanol Nutrition 0.000 claims description 16
- 239000002504 physiological saline solution Substances 0.000 claims description 15
- 235000014347 soups Nutrition 0.000 claims description 10
- 238000011161 development Methods 0.000 claims description 9
- 238000002224 dissection Methods 0.000 claims description 9
- 238000001035 drying Methods 0.000 claims description 9
- 239000012530 fluid Substances 0.000 claims description 5
- 238000010521 absorption reaction Methods 0.000 claims description 3
- 239000000284 extract Substances 0.000 claims description 3
- 230000002093 peripheral effect Effects 0.000 claims description 3
- 238000000605 extraction Methods 0.000 claims description 2
- 239000007924 injection Substances 0.000 claims description 2
- 238000002347 injection Methods 0.000 claims description 2
- 235000013601 eggs Nutrition 0.000 abstract description 38
- 230000013020 embryo development Effects 0.000 abstract description 10
- 239000000725 suspension Substances 0.000 abstract description 8
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 abstract description 5
- 241001518462 Cubiceps gracilis Species 0.000 abstract description 2
- 238000003756 stirring Methods 0.000 abstract description 2
- 241000594009 Phoxinus phoxinus Species 0.000 description 8
- 230000018109 developmental process Effects 0.000 description 8
- WSFSSNUMVMOOMR-UHFFFAOYSA-N Formaldehyde Chemical compound O=C WSFSSNUMVMOOMR-UHFFFAOYSA-N 0.000 description 6
- 241001519451 Abramis brama Species 0.000 description 5
- 241000404975 Synchiropus splendidus Species 0.000 description 5
- 239000004833 fish glue Substances 0.000 description 5
- 210000001647 gastrula Anatomy 0.000 description 5
- 239000000049 pigment Substances 0.000 description 4
- 238000007796 conventional method Methods 0.000 description 3
- 238000000151 deposition Methods 0.000 description 3
- 210000003462 vein Anatomy 0.000 description 3
- XUMBMVFBXHLACL-UHFFFAOYSA-N Melanin Chemical compound O=C1C(=O)C(C2=CNC3=C(C(C(=O)C4=C32)=O)C)=C2C4=CNC2=C1C XUMBMVFBXHLACL-UHFFFAOYSA-N 0.000 description 2
- 241000356847 Otolithes Species 0.000 description 2
- 239000002775 capsule Substances 0.000 description 2
- 230000018044 dehydration Effects 0.000 description 2
- 238000006297 dehydration reaction Methods 0.000 description 2
- 210000003128 head Anatomy 0.000 description 2
- 230000000968 intestinal effect Effects 0.000 description 2
- 230000002842 otolith Effects 0.000 description 2
- 210000001265 otolithic membrane Anatomy 0.000 description 2
- 210000000006 pectoral fin Anatomy 0.000 description 2
- 102000002322 Egg Proteins Human genes 0.000 description 1
- 108010000912 Egg Proteins Proteins 0.000 description 1
- 210000000436 anus Anatomy 0.000 description 1
- 210000004204 blood vessel Anatomy 0.000 description 1
- 230000008602 contraction Effects 0.000 description 1
- 238000004042 decolorization Methods 0.000 description 1
- 238000013461 design Methods 0.000 description 1
- 238000002474 experimental method Methods 0.000 description 1
- 210000000936 intestine Anatomy 0.000 description 1
- 210000005036 nerve Anatomy 0.000 description 1
- 210000004681 ovum Anatomy 0.000 description 1
- 230000000149 penetrating effect Effects 0.000 description 1
- 238000004321 preservation Methods 0.000 description 1
- 238000003672 processing method Methods 0.000 description 1
- 238000011160 research Methods 0.000 description 1
- 239000007787 solid Substances 0.000 description 1
- 239000000243 solution Substances 0.000 description 1
- 210000001519 tissue Anatomy 0.000 description 1
Classifications
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01K—ANIMAL HUSBANDRY; AVICULTURE; APICULTURE; PISCICULTURE; FISHING; REARING OR BREEDING ANIMALS, NOT OTHERWISE PROVIDED FOR; NEW BREEDS OF ANIMALS
- A01K61/00—Culture of aquatic animals
- A01K61/10—Culture of aquatic animals of fish
-
- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01K—ANIMAL HUSBANDRY; AVICULTURE; APICULTURE; PISCICULTURE; FISHING; REARING OR BREEDING ANIMALS, NOT OTHERWISE PROVIDED FOR; NEW BREEDS OF ANIMALS
- A01K61/00—Culture of aquatic animals
- A01K61/10—Culture of aquatic animals of fish
- A01K61/17—Hatching, e.g. incubators
-
- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02A—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE
- Y02A40/00—Adaptation technologies in agriculture, forestry, livestock or agroalimentary production
- Y02A40/80—Adaptation technologies in agriculture, forestry, livestock or agroalimentary production in fisheries management
- Y02A40/81—Aquaculture, e.g. of fish
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- Life Sciences & Earth Sciences (AREA)
- Environmental Sciences (AREA)
- Marine Sciences & Fisheries (AREA)
- Zoology (AREA)
- Animal Husbandry (AREA)
- Biodiversity & Conservation Biology (AREA)
- Investigating Or Analysing Biological Materials (AREA)
- Endoscopes (AREA)
Abstract
The invention discloses a kind of production Drifting egg fish early stage different developmental phases form microscopy observation methods, it is characterised in that is made of Larval morphologic observation method after developmental morphology observation method between drift fish-egg after fertilization to developmental morphology observation method during tail bud, muscle effects phase to membrane early period and membrane.The method of the present invention can solve because drift fish fish-egg after fertilization to during tail bud because embryo is smaller, egg membrane is excessive, in fertilized eggs water in suspension, embryonic suspension in being flowed in pairs in egg membrane, be not easy to find best shooting angle problem；Secondly between solving muscle effects phase to membrane early period, embryo shrinks in egg membrane, and embryo stirs, and is not easy to obtain complete embryo image problem；It is finally that prelarva is easily mostly dynamic after solving membrane, can not fix frame problem of taking pictures.It was verified that for drift property fish-egg or prelarva microexamination, taking pictures, adjustable angle, embryonic development picture easily capture, and prelarva seldom decolourizes, is dehydrated, shooting effect is fresh and alive, details is complete.
Description
Technical field
The present invention relates to a kind of production Drifting egg fish early stage different developmental phases form microscopy observation methods, belong to fish
Morphological observation field.
Background technology
The widely distributed inland rivers water system of fish for producing Drifting egg, by being research to its fish-egg, the photography of prelarva live body
One important means of fish early life history.Fix preservation fish-egg and prelarva by fixer is to observe its morphological development situation
There is observation, ineffective problem of taking pictures, need also exist for often replacing fixer, step in currently used method, this method
The problems such as complexity is easily destroyed sample structure during depositing, withdrawing, and fixer penetrating odor is big.Drift fish-egg fertilized eggs
Entire body is transparent, directly can dissect microscopic observation embryonic development situation.But because there are drift fish-eggs because embryo is smaller, egg membrane mistake
Greatly, in fertilized eggs water in suspension, embryonic suspension in egg membrane and caused by flow in pairs, be not easy to find best shooting angle；Secondly
After embryonic development to muscle effects phase, because of embryo motion, portion of tissue is often affixed on endocardial wall, is not easy to obtain complete embryo image；
Live body prelarva is easily mostly dynamic, be not easy to fix picture catching development details the problems such as.
Therefore, there is an urgent need for design a kind of production Drifting egg fish early stage different developmental phases form microscopy observation method.
Invention content
The present invention is for production drift fish-egg fish early development in the process because drift fish-egg is because embryo is smaller, egg membrane mistake
Greatly, in fertilized eggs water in suspension, embryonic suspension in egg membrane and caused by flow in pairs, be not easy to find best shooting angle；Secondly
After embryonic development to muscle effects phase, embryo shrinks in egg membrane, is not easy to obtain complete embryo image；Live body prelarva is easily mostly dynamic,
Be not easy to fix picture catching development details the problems such as and a kind of production Drifting egg fish early stage different developmental phases form for designing
Microscopy observation method.
To achieve the above object, technical scheme is as follows：
A kind of production Drifting egg fish early stage different developmental phases form microscopy observation method, it is characterised in that：
It is observed by developmental morphology between fish-egg after fertilization to developmental morphology observation method during tail bud, muscle effects phase to membrane early period
Larval morphologic observation method forms after method and membrane；
The fish-egg after fertilization to developmental morphology observation method during tail bud is：
（1）It takes after fertilization to one, fish-egg during tail bud with suction pipe or small soup ladle, puts to clean, dry culture dish central area
Domain；
（2）Moisture between being contacted with culture dish with the small-sized strip sponge exhaustion fish-egg of drying, enables fish-egg to be stabilized in culture dish
On；
（3）The above-mentioned culture dish for being equipped with fish-egg is placed under dissection endoscope objective lens, fish-egg is stirred extremely with the small-sized strip sponge of drying
Required observation or shooting angle；
（4）Anatomical lens is adjusted, observation or picture of taking pictures are obtained；Observation is further completed in 5 minutes or is taken pictures；
The small-sized strip sponge port area is no more than 0.1cm2。
Phasic development morphologic observation method is between the muscle effects phase to membrane early period：
（1）The muscle effects phase is taken to be put to one, the fish-egg of membrane preliminary stage to clean, desiccation culture ware with suction pipe or small soup ladle
Middle section；
（2）Moisture between being contacted with culture dish with the small-sized strip sponge exhaustion fish-egg of drying, enables fish-egg to be stabilized in culture dish
On；The small-sized strip sponge port area is no more than 0.1cm2。
（3）The above-mentioned culture dish for being equipped with fish-egg is placed under dissection endoscope objective lens, the dissection micro- multiple of sem observation is adjusted, uses
Line needle punctures fish-egg film in opposite directions from fish-egg left and right sides respectively, and the vertical height punctured a little is that fish-egg is contacted with culture dish at 3/4；Institute
The length for stating line needle is 3-5cm；
（4）After embryo flows out film, on dropper drop 0.1-0.2ml physiological saline to embryo；
（5）With 10ml syringes along embryo's peripheral liquid is extracted, until egg membrane is sucked in syringe needle, egg membrane is removed, if liquid
Body is drained and egg membrane is not sucked also, then adds 0.1-0.2ml physiological saline with dropper, extracts again, can be repeated several times, until injection
Egg membrane is sucked in device syringe needle, removes egg membrane；
（6）With 10ml syringes toward the ethyl alcohol for 50% concentration of 0.05-0.1ml of dripping on striping embryo, after standing 1-2 minutes, then
With 10ml syringes toward the ethyl alcohol for 50% concentration of 0.04-0.05ml of dripping on striping embryo, 1-2 minutes are stood；
（7）It controls 0.1-0.2ml liquid and covers embryo, the control method is：If being drawn with 10ml syringes when liquid is more
Surplus liquid, if being supplemented with physiological saline when fluid low；
（8）Mobile culture dish is located to embryo in anatomical lens field of view, adjusts anatomical lens multiple, obtains observation or picture of taking pictures
Face；Observation is further completed in 5 minutes or is taken pictures.
Larval morphologic observation method is after the membrane：
（1）One tail of prelarva is taken with suction pipe or small soup ladle, is put to clean, dry culture dish middle section；
（2）Control prelarva is added or drawn with 10ml syringes is no more than 1ml with culture dish contact portion moisture；
（3）The ethyl alcohol for 90% concentration of 0.05-0.1ml of being dripped around prelarva with 10ml syringes after standing 2-3 minutes, then is used
10ml syringes drip the ethyl alcohol of 50% concentration of 0.05-0.1ml around prelarva, stand 3-4 minutes；
（4）It controls 0.3-0.4ml liquid and covers prelarva；The control methods are：More with the absorption of 10ml syringes when if liquid is more
Extraction raffinate body, if being supplemented with physiological saline when fluid low；
（5）Mobile culture dish to prelarva is located in anatomical lens field of view, adjusts anatomical lens multiple, obtains observation or picture of taking pictures
Face further completes observation or takes pictures in 5 minutes；If prelarva eye lens is reflective, dripped on prelarva eye with 10ml syringes
0.04-0.1ml physiological saline stands 1 minute, adjusts anatomical lens multiple, acquisition is observed or picture of taking pictures, further 5 minutes
Interior completion observation is taken pictures.
Advantageous effect：
1, the method for the present invention can solve because drift fish fish-egg after fertilization to during tail bud because embryo is smaller, egg membrane is excessive, fertilization
In ovum water in suspension, embryonic suspension in being flowed in pairs in egg membrane, be not easy to find best shooting angle problem；Secondly muscle effects are solved
Between phase to membrane early period, embryo shrinks in egg membrane, and easily stirs, and is not easy to obtain complete embryo image problem；It is finally that solution has decided
Prelarva is easily mostly dynamic after film, and can not fix frame problem of taking pictures.It was verified that the method for the present invention is used for drift property fish-egg or prelarva
Microexamination is taken pictures, and adjustable angle, embryonic development picture easily capture, and prelarva seldom decolourizes, is dehydrated, shooting effect is fresh and alive, thin
Section is complete.
2, compared with generally preserving observation using fixer, the method for the present invention can clearly observe the embryo of fish-egg embryo
Whether ring, embryonic shield, eye capsule, statocyst, eye lens, otolith occur；Whether blastopore is closed；Whether muscle segment forms；Whether muscle starts
The details such as contraction；The fin fold of prelarva, pigment, intestines, anus, blood vessel start to occur；The details such as a few rooms of fish glue, muscle segment number.And it uses solid
Determine liquid and preserve observation sample integrally to decolourize seriously, many details can not be observed clear, and easily lead to sample in the process of depositing, withdrawing
Damage, can not ensure sample integrity.
3, the present invention uses different microexamination processing methods for different stage of development features, will not lead to sample
Dehydration, color throw, clear and intuitive can observe production Drifting egg fish early stage different developmental phases details of morphology variation,
Can be to distinguish the type of fish or judge early development specific period provide important evidence.
4, in the muscle effects phase of the present invention to membrane preliminary stage developmental morphology observation method during striping, by using
Line needle punctures fish-egg film in opposite directions from fertilized eggs left and right sides respectively, and control punctures height a little, can be effectively completely by fish-egg film
Removal, makes embryo completely show, can clearly observe and obtain entire embryonic development form；
5, larval stages use the second of 50% concentration by largely groping after the muscle effects phase to membrane preliminary stage and membrane
Alcohol and the ethyl alcohol of 90% concentration handle sample, while determining dosage and action time, can effectively avoid embryo or son
Fish struggle deformation or dehydration and decolorization ensure that embryo or prelarva keep natural form, greatly improve the accuracy of observation, make
The original form that result more realistically reflects embryo or Larval must be observed.
Figure of description
Fig. 1 Wei Wen Minnow blastopores close phase embryonic development aspect graph；
Fig. 2 is Wen Minnow heartbeat phase embryonic development aspect graphs Xia different observation methods；
Fig. 3 is to observe mandarin fish alevin stage developmental morphology figure using distinct methods；
Fig. 4 is to observe bream alevin stage developmental morphology figure using distinct methods.
Specific embodiment
Embodiment 1
A kind of production Drifting egg fish early stage different developmental phases form microscopy observation method, closing Qi Wen Minnow fish-eggs with blastopore is
Example, observation method are as follows：
（1）One, blastopore closing Qi Wen Minnow fish-eggs are taken with suction pipe or small soup ladle, are put to clean, dry culture dish middle section；
（2）Moisture between being contacted with culture dish with the small-sized strip sponge exhaustion fish-egg of drying, enables fish-egg to be stabilized in culture dish
On；
（3）The above-mentioned culture dish for being equipped with fish-egg is placed under dissection endoscope objective lens, fish-egg is stirred extremely with the small-sized strip sponge of drying
Required observation or shooting angle；
（4）Anatomical lens is adjusted, observation or picture of taking pictures are obtained, observation is further completed in 5 minutes or is taken pictures；
The small-sized strip sponge port area is no more than 0.1cm2。
Embodiment 2
A kind of production Drifting egg fish early stage different developmental phases form microscopy observation method is with heartbeat Qi Wen Minnow fish-eggs
Example, observation method are as follows：
（1）One, heartbeat phase kiss Minnow fish-egg is taken with suction pipe or small soup ladle, is put to clean, desiccation culture ware middle section；
（2）Moisture between being contacted with culture dish with the small-sized strip sponge exhaustion fish-egg of drying, enables fish-egg to be stabilized in culture dish
On；The small-sized strip sponge port area is no more than 0.1cm2。
（3）The above-mentioned culture dish for being equipped with fish-egg is placed under dissection endoscope objective lens, the dissection micro- multiple of sem observation is adjusted, uses
Line needle punctures fish-egg film in opposite directions from fish-egg left and right sides respectively, and the vertical height punctured a little is that fish-egg is contacted with culture dish at 3/4；Institute
The length for stating line needle is 3cm；
（4）After embryo flows out film, on dropper drop 0.15ml physiological saline to embryo；
（5）With 10ml syringes along embryo's peripheral liquid is extracted, until egg membrane is sucked in syringe needle, egg membrane is removed, if liquid
Body is drained and egg membrane is not sucked also, then adds 0.15ml physiological saline with dropper, extracts again, can be repeated several times, until syringe
Egg membrane is sucked in syringe needle, removes egg membrane；
（6）With 10ml syringes toward the ethyl alcohol for 50% concentration of 0.08ml of dripping on striping embryo, after standing 1 minute, then 10ml is used
Syringe stands 1 minute toward the ethyl alcohol for 50% concentration of 0.04ml of dripping on striping embryo；
（7）It controls 0.15ml liquid and covers embryo, the control method is：Extra with the absorption of 10ml syringes when if liquid is more
Liquid, if being supplemented with physiological saline when fluid low；
（8）Mobile culture dish is located to embryo in anatomical lens field of view, adjusts anatomical lens multiple, obtains observation or picture of taking pictures
Face further completes observation or takes pictures in 5 minutes.
Embodiment 3
A kind of production Drifting egg fish early stage different developmental phases form microscopy observation method, by taking mandarin fish prelarva as an example, watcher
Method is as follows：
（1）One tail of mandarin fish prelarva is taken with suction pipe or small soup ladle, is put to clean, dry culture dish middle section；
（2）It is 0.5ml that control prelarva is added or drawn with 10ml syringes with culture dish contact portion moisture；
（3）The ethyl alcohol for 90% concentration of 0.08ml of being dripped around prelarva with 10ml syringes, standing are noted after 2 minutes, then with 10ml
Emitter is dripped the ethyl alcohol of 50% concentration of 0.08ml around prelarva, stands 3 minutes；
（4）It controls 0.35ml liquid and covers prelarva；The control methods are：10ml syringes draw surplus liquid；
（5）Mobile culture dish to prelarva is located in anatomical lens field of view, adjusts anatomical lens multiple, obtains observation or picture of taking pictures
Face further completes observation or takes pictures in 5 minutes；If prelarva eye lens is reflective, dripped on prelarva eye with 10ml syringes
0.07ml physiological saline stands 1 minute, adjusts anatomical lens multiple, obtains observation or picture of taking pictures, further complete in 5 minutes
At observing or take pictures.
Embodiment 4
A kind of production Drifting egg fish early stage different developmental phases form microscopy observation method, by taking bream prelarva as an example, watcher
Method is as follows：
（1）One tail of bream prelarva is taken with suction pipe or small soup ladle, is put to clean, dry culture dish middle section；
（2）Control prelarva and culture dish contact portion moisture 0.3ml are added or drawn with 10ml syringes；
（3）The ethyl alcohol for 90% concentration of 0.05ml of being dripped around prelarva with 10ml syringes after standing 2.5 minutes, then uses 10ml
Syringe drips the ethyl alcohol of 50% concentration of 0.06ml around prelarva, stands 3 minutes；
（4）It controls 0.3ml liquid and covers prelarva；The control methods are：Surplus liquid is drawn with 10ml syringes；
（5）Mobile culture dish adjusts anatomical lens multiple until prelarva is located in anatomical lens field of view, obtains observation or picture of taking pictures
Face further completes observation or takes pictures in 5 minutes；If prelarva eye lens is reflective, dripped on prelarva eye with 10ml syringes
0.06ml physiological saline stands 1 minute, adjusts anatomical lens multiple, obtains observation or picture of taking pictures, further complete in 5 minutes
At observing or take pictures.
Specific experiment effect：
Fig. 1 is that the microexamination of 1 observation method Wen Minnow fish-eggs of embodiment is taken pictures figure, and embryonic development blastopore closes phase, nerve channel
It is formed.
Fig. 2 is that the microexamination of 2 observation method Wen Minnow fish-eggs of embodiment is taken pictures figure, and embryonic development enters heartbeat
Phase, heart former base occur, and heart beats, eye capsule statocyst is high-visible, occurs otolith in statocyst, at this time muscle segment about 32
It is right；
Conventional method microexamination is taken pictures figure, by embryo can in film significantly this experience of shift position, can be substantially
Judge development enter the heartbeat phase, but embryo head, tail portion details not it is observed that.Due to there is no striping to fix, even if even
You have found that embryo head, tail portion are moved in field of view, and flash across, and detail can not capture, and cannot make very
It is good to judge and observe.
Fig. 3 is that the microexamination of the mandarin fish prelarva of 3 observation method of embodiment is taken pictures figure, when jaw tooth occurs, front and back nucleus point
Boundary is apparent, occurs spine on the gill cover, and the hard spine of pectoral fin is formed, and occurs the starlike melanin of row at tail vein；
Using formaldehyde fixing means（Conventional method）Mandarin fish prelarva microexamination is taken pictures figure, and whole decoloration is serious, can not observe whether there is or not
Jaw tooth, front and back gill cover boundary unobvious, can not judge the gill cover, whether the spine of pectoral fin forms.
Fig. 4 is that the microexamination of the bream prelarva of 3 observation method of embodiment is taken pictures figure, and body is slender, and two Room of fish glue is high-visible, the heart
Dirty position is in cerise, and dorsal fin pleat slightly swells, starts to break up, and side lower part is in blue veins one along intestinal tube to caudal fold rear portion pigment from fish glue
One big pigment flower, caudal myotome 20 are also arranged at item, caudal fold lower part.
Using formaldehyde fixing means（Conventional method）Bream prelarva microexamination is taken pictures figure, and prelarva has damaged because depositing, withdrawing process
Tail portion, a fish glue only Room is as it can be seen that prelarva decoloration seriously, is not easy to observe that side lower part is in along intestinal tube to caudal fold rear portion pigment from fish glue
One, blue veins, it is clear that muscle segment is not easy number.
Claims (5)
1. a kind of production Drifting egg fish early stage different developmental phases form microscopy observation method, it is characterised in that：
It is observed by developmental morphology between fish-egg after fertilization to developmental morphology observation method during tail bud, muscle effects phase to membrane early period
Larval morphologic observation method forms after method and membrane.
2. a kind of production Drifting egg fish early stage different developmental phases form microscopy observation method according to claim 1,
It is characterized in that：
The fish-egg after fertilization to developmental morphology observation method during tail bud is：
（1）It takes after fertilization to one, fish-egg during tail bud with suction pipe or small soup ladle, puts to clean, dry culture dish central area
Domain；
（2）Moisture between being contacted with culture dish with the small-sized strip sponge exhaustion fish-egg of drying, enables fish-egg to be stabilized in culture dish
On；
（3）The above-mentioned culture dish for being equipped with fish-egg is placed under dissection endoscope objective lens, fish-egg is stirred extremely with the small-sized strip sponge of drying
Required observation or shooting angle；
（4）Anatomical lens is adjusted, observation or picture of taking pictures are obtained；
The small-sized strip sponge port area is no more than 0.1cm2。
3. a kind of production Drifting egg fish early stage different developmental phases form microscopy observation method according to claim 1,
It is characterized in that：
Phasic development morphologic observation method is between the muscle effects phase to membrane early period：
（1）The muscle effects phase is taken to be put to one, the fish-egg of membrane preliminary stage to clean, desiccation culture ware with suction pipe or small soup ladle
Middle section；
（2）Moisture between being contacted with culture dish with the small-sized strip sponge exhaustion fish-egg of drying, enables fish-egg to be stabilized in culture dish
On；The small-sized strip sponge port area is no more than 0.1cm2；
（3）The above-mentioned culture dish for being equipped with fish-egg is placed under dissection endoscope objective lens, the dissection micro- multiple of sem observation is adjusted, using line needle
Fish-egg film is punctured in opposite directions from fish-egg left and right sides respectively, and the vertical height punctured a little is that fish-egg is contacted with culture dish at 3/4；The line
The length of needle is 3-5cm；
（4）After embryo flows out film, on dropper drop 0.1-0.2ml physiological saline to embryo；
（5）With 10ml syringes along embryo's peripheral liquid is extracted, until egg membrane is sucked in syringe needle, egg membrane is removed, if liquid
Body is drained and egg membrane is not sucked also, then adds 0.1-0.2ml physiological saline with dropper, extracts again, can be repeated several times, until injection
Egg membrane is sucked in device syringe needle, removes egg membrane；
（6）With 10ml syringes toward the ethyl alcohol for 50% concentration of 0.05-0.1ml of dripping on striping embryo, after standing 1-2 minutes, then
With 10ml syringes toward the ethyl alcohol for 50% concentration of 0.04-0.05ml of dripping on striping embryo, 1-2 minutes are stood；
（8）It controls 0.1-0.2ml liquid and covers embryo, the control method is：If being drawn with 10ml syringes when liquid is more
Surplus liquid, if being supplemented with physiological saline when fluid low；
（9）Mobile culture dish is located to embryo in anatomical lens field of view, adjusts anatomical lens multiple, obtains observation or picture of taking pictures
Face.
4. a kind of production Drifting egg fish early stage different developmental phases form microscopy observation method according to claim 1,
It is characterized in that：
Larval morphologic observation method is after membrane：
（1）One tail of prelarva is taken with suction pipe or small soup ladle, is put to clean, dry culture dish middle section；
（2）Control prelarva is added or drawn with 10ml syringes is no more than 1ml with culture dish contact portion moisture；
（3）The ethyl alcohol for 90% concentration of 0.05-0.1ml of being dripped around prelarva with 10ml syringes after standing 2-3 minutes, then is used
10ml syringes drip the ethyl alcohol of 50% concentration of 0.05-0.1ml around prelarva, stand 3-4 minutes；
（4）It controls 0.3-0.4ml liquid and covers prelarva；The control methods are：More with the absorption of 10ml syringes when if liquid is more
Extraction raffinate body, if being supplemented with physiological saline when fluid low；
（5）Mobile culture dish to prelarva is located in anatomical lens field of view, adjusts anatomical lens multiple, obtains observation or picture of taking pictures
Face further completes observation or takes pictures in 5 minutes；If prelarva eye lens is reflective, dripped on prelarva eye with 10ml syringes
0.04-0.1ml physiological saline stands 1 minute, adjusts anatomical lens multiple, obtains observation or picture of taking pictures.
5. according to a kind of any production Drifting egg fish early stage different developmental phases form microexaminations of claim 2-4
Method, it is characterised in that the observation or picture of taking pictures are completed in 5 minutes.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201810460288.9A CN108651337B (en) | 2018-05-15 | 2018-05-15 | Method for observing early development morphology of Siniperca mandarin fish with bleeded eggs in gobiocypyristoides |
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