CN105571916B - A kind of tissue section method of acipenser dabryanus fertilized eggs - Google Patents
A kind of tissue section method of acipenser dabryanus fertilized eggs Download PDFInfo
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Abstract
The present invention relates to microscopic tissue sections technologies, are exactly specifically a kind of tissue section method of acipenser dabryanus fertilized eggs.Mainly fixed, be dehydrated by material, is transparent, waxdip, embedding, slice and dyeing seven steps form；The material fixes the formaldehyde fixer that the fixer used is 10% for Bouin ' s fixers or mass fraction；The dehydration total time is 10 20h.The present invention is directed to sturgeon oosperm and belongs to viscid egg, the features such as Yolk is abundant, the tough and tensile densification of egg membrane, perivitelline is narrow, the Serial tissue sections method suitable for sturgeons fertilized eggs of complete set is established, fertilized eggs are effectively handled and produces preferable paraffin section.
Description
Technical field
The present invention relates to microscopic tissue sections technologies, are exactly specifically a kind of histotomy side of acipenser dabryanus fertilized eggs
Method.
Background technology
Acipenser dabryanus (Acipenser dabryanus) is also known as acipenser dabryanus and Sha Lazi, is the distinctive fresh water in China colonization property
Acipenseridae (Acipenseridae) sturgeon category fish are distributed in Upper Yangtze River and Jinsha jiang River downstream (auspicious magnificent 1994) of fourth, national I grade of weight
Point protection wild animal and Upper Yangtze River level-one protect fish (Liu Jun 2004), International Union for Conservation of Nature and Natural Resources (World in a hurry
Conservation Union, IUCN) pole danger grade (critically endangered, CR) species (Du Jun etc. 2009), it is international
Endangered animal and plant kind trade pact (Convention on International Trade in Endangered Species
Of Wild Fauna and Flora, CITES) appendix II protection species, the rare Endemic fish of Upper Yangtze River is national to be protected naturally
Protect one of the main protected object in area.In recent years, due to the deterioration of Upper Yangtze River rivers ecological environment, the exploitation of step hydropower station,
Natively less acipenser dabryanus resource further declines, and is difficult now to find, be more difficult to capture.In order to preferably protect its biology money
Population is restored in source, stablizes ecological environment, and the reproductive study of sturgeons seems increasingly important.In addition ensureing population quantity
In the case of, acipenser dabryanus fish-egg can be used as the important source material of caviar, have higher economic value, to economic development and raising
People's living standard has certain contribution.Therefore, research acipenser dabryanus fertilized eggs have very positive effect.
For the histotomy technology of fish oosperm, mainly using the fertilized eggs of freshwater fish as experiment material, and
And be mainly the cyprinid fish chosen, such as the fertilized eggs microtomy of grass carp and crucian, also there is less seawater fish fertilization
Ovum microtomy, such as left-eyed flounder.And the relevant technologies for sturgeons fertilized eggs are there is not yet report.The side of freshwater fish
Method is striping after fixing its fertilized eggs Bouin ' s fixers or Smith fixers, according to general tissue section method
By Gradient elution using ethanol, dimethylbenzene is transparent, paraffin embedding and etc. after, carry out serial section, paraffin section is made.And seawater
Fish are to puncture egg membrane and agar embedding step with reference on the basis of general dicing method through having gone before dewatering, enhance
Dehydration and positioning.
On acipenser dabryanus, the fertilized eggs of its different development stage are handled through row using above two method, the feelings of appearance
Condition is often histotomy and is crushed, and Yolk is fallen, and egg membrane fracture, separation cannot be obtained than being more completely sliced.It is main
The reason is that acipenser dabryanus ovum ovum diameter is larger, the tough and tensile densification of egg membrane, it is fixed after ovum compared with most of freshwater fishes its perivitelline it is non-
It is often narrow, cannot completely striping, and since ovum week covers one layer of thicker mucous membrane, in dehydration, transparent, waxdip process
In it is abundant not as good as the processing of other fish, in the difference of the main component of fertilized eggs different development stage, cause to be sliced successfully difficulty
Greatly.
Invention content
Present invention solves the technical problem that be that acipenser dabryanus ovum ovum diameter is larger, the tough and tensile densification of egg membrane, it is fixed after ovum with mostly
Number freshwater fishes are very narrow compared to its perivitelline, cannot completely striping, and due to belonging to viscid egg, ovum week covering one
The thicker mucous membrane of layer, dehydration, it is transparent, waxdip during, general fish oosperm group abundant not as good as the processing of other fish
It knits dicing method and is not suitable for sturgeons, slice success rate is relatively low.
To solve the above problems, our experiments Jing Guo system, include the trial of method, the optimization of condition, the tune of time
It is whole, and a large amount of repetition experiment, the Serial tissue sections method suitable for sturgeons fertilized eggs of complete set is established, is built
The method and condition for having found suitable sturgeons fertilized eggs histotomy is directed to sturgeon oosperm and belongs to viscid egg, Yolk
It is abundant, the features such as the tough and tensile densification of egg membrane, perivitelline is narrow, effectively handles fertilized eggs and produce preferable paraffin section.
Technical scheme of the present invention mainly fixed by material, is dehydrated, is transparent, waxdip, embedding, seven steps of slice and dyeing
Composition；The material fixes the formaldehyde fixer that the fixer used is 10% for Bouin ' s fixers or mass fraction；It is described
Dewatering time is 1-2h.The transparent step is after transparent liquid is handled, then passes through terpinol processing 4-5h and pine for the first time respectively
Oleyl alcohol handles 5-7h for the second time, until the transparent state of sturgeon ovum；The dehydration total time is 10-20h；The waxdip time is
16-30h。
The material fixing step is that fresh sturgeon ovum is put into Bouin ' the s fixers or volume of 40-60 times of volume
After fixing 20-30 hours in the formaldehyde fixer that score is 10%, it is that 70% ethyl alcohol rinses repeatedly with volume fraction, then preserves
In the ethyl alcohol that volume fraction is 70%, the alcohol that a volume fraction is 70% is replaced every two days, is continuously preserved afterwards three times standby
With.
Preferably, the material fixing step is that fresh sturgeon ovum is put into Bouin ' the s fixers of 50 times of volumes
After fixing 24 hours in middle fixation or 10% formaldehyde fixer, it is stored in after being rinsed repeatedly with 70% ethyl alcohol in 70% ethyl alcohol,
70% alcohol is replaced every two days continuously three times fully to wash away fixer, to save backup.
The dehydration is that the sturgeon ovum stored in 70% ethyl alcohol is directly immersed in 80% ethanol dehydration primary, the time
2h, and pass sequentially through 90% ethanol dehydration 2h, 95% ethanol dehydration twice, each 1-2h, 100% ethanol dehydration
Twice, each 1h.
The transparent step be first by through dewatered sturgeon ovum change to absolute ethyl alcohol mix in equal volume with terpinol it is saturating
Bright liquid or terpinol：Dimethylbenzene：Absolute ethyl alcohol=4：1：1-3h is handled in 5 mixed transparent transition fluid, is then passed through respectively again
Crossing terpinol, processing 4-5h and terpinol handle 5-7h for the second time for the first time, until the transparent state of sturgeon ovum.
The waxdip step is that the sturgeon ovum after transparent processing is first changed to the waxdip that terpinol mixes in equal volume with paraffin
1-2h is handled in liquid, then passing through paraffin respectively again, processing 5-8h and paraffin handle 10-20h for the second time for the first time.
The embedding step is to be wrapped the sturgeon ovum conventional organization embedding method after the completion of waxdip with carton
It buries, paraffin mass is made.
The slice is by the continuous paraffin section of slicer, by opening up piece, patch, baking piece, by Soviet Union with staining procedure
Another name for Yihong (HE) is dyed, and last microexamination is taken pictures.The specific steps are with microtome, slice thickness is controlled in 5 μm of -7 μ
M, through being extended piece in 35 DEG C of -38 DEG C of water-baths, after dragging for piece with the glass slide for being coated with albumen glycerine, in 35 DEG C -38 DEG C of baking oven
In carry out baking piece.
Haematoxylin ＆ eosin (HE) decoration method is as follows：First use first time dimethylbenzene processing 10-15min, second
It is 1 that dimethylbenzene, which handles 10-13min, uses volume ratio again,:1 dimethylbenzene：Absolute ethyl alcohol handles 2min, the ethyl alcohol of first time 100%
Handle 2min, second 100% of alcohol treatment 2min, for the first time 95% alcohol treatment 2min, second 95% of ethyl alcohol
2min, 80% alcohol treatment 2min, 70% alcohol treatment 2min, hematoxylin dyeing 20min, tap water is handled to rinse
3min, 1% hydrochloride alcohol solution processing 5s, AMMONIA TREATMENT 15s after washing, wash again, 70% alcohol treatment 3min, 80%
Alcohol treatment 3min, 90% alcohol treatment 3min, eosin stains 30s, 95% alcohol treatment 4min, for the first time 100%
Alcohol treatment 4min, second 100% alcohol treatment 4min, with volume ratio be 1:1 absolute ethyl alcohol：Dimethylbenzene processing
5min, first time dimethylbenzene processing 5min, second of dimethylbenzene processing 5min are allowed to transparent, then use neutral gum mounting.
The acipenser dabryanus fish fertilized egg histotomy prepared using the method for the present invention it can be seen from attached drawing 1-4, egg membrane are complete
Whole, completely in flakes, no incomplete, overlapping, institutional framework is clear, well arranged, and cell boundary line understands that staining versus is bright for histotomy
It is aobvious, have no grey dye.
The beneficial effects of the invention are as follows：
1. the present invention establishes the method and condition of suitable sturgeons fertilized eggs histotomy, belong to for sturgeon oosperm
The features such as viscid egg, Yolk is abundant, the tough and tensile densification of egg membrane, perivitelline is narrow, the present invention can effectively handle fertilized eggs simultaneously
Produce preferable paraffin section.There is scholar to think that the fixed fertilized eggs of Bouin ' s fixers will appear Yolk hard and crisp
The case where, so that it cannot serial section, some scholars think fixed effect ratio Bouin ' the s fixers of Smith fixers
It is good, it is not easy to so that fertilized eggs is hardened crisp, but in the present invention, use Bouin ' s fixers or 10% formaldehyde fixer to handle
Left and right will not make Yolk be hardened crisp for 24 hours, being capable of serial section.Therefore, the present invention can be reduced using Smith fixer bands
The troublesome operation step come.
2. the present invention passes through a large amount of explorations to dewatering time, it is determined that best dewatering time, solve it is most of by
The problem of cannot being fully dehydrated when smart ovum histotomy.Since sturgeons fertilized eggs belong to viscid egg, periphery has one layer thicker
Mucous membrane, its level-one egg membrane and the tough and tensile densification of two level egg membrane in addition, if dewatering time is not set, can cause dehydration when hold
Easy drastically catastrophic collapse, causes ovum week uneven, influences to be sliced.Conventional method is that the method for peeling egg membrane off is asked to solve this
Topic, but sturgeons fertilized eggs perivitelline is very narrow, cannot divest, and also has the method by being punched on egg membrane with dissecting needle
It solves, but increases operating procedure, and the present invention is by the control of dewatering time, without carrying out special place to egg membrane
Reason does not only reach the purpose being dehydrated completely, and the integrality of the egg membrane retained.
3. experiment sturgeons fertilized eggs multiple periods through having gone identical processing method, be obtained for complete
Histotomy, repetitive rate is high, and effect is good, therefore the present invention is suitable for the histotomy in sturgeons fertilized eggs each period.
4. the present invention is easy to operate, rationally proper, it is not only applicable to the histotomy of sturgeons fertilized eggs, is also applied for gluing
Property ovum, the fertilized eggs histotomy of other megalecithal fish are applied widely.
Description of the drawings
Fig. 1：The single complete ovum that the cleavage stage histotomy of acipenser dabryanus fertilized eggs obtains is carried out using the embodiment of the present invention 1
Slice；
Fig. 2：The egg membrane amplification that the cleavage stage histotomy of acipenser dabryanus fertilized eggs obtains is carried out using the embodiment of the present invention 1
According to；
Fig. 3：The histotomy slice map in the primitive gut period of acipenser dabryanus fertilized eggs is carried out using the embodiment of the present invention 2；
Fig. 4：Acipenser dabryanus fertilized eggs phase histotomy figure is carried out using the embodiment of the present invention 3；
Fig. 5：Comparison study embodiment 1 carries out acipenser dabryanus fertilized eggs gastrul stage histotomy figure；
Fig. 6：Comparison study embodiment 2 carries out acipenser dabryanus fertilized eggs fertilized eggs phase histotomy egg membrane figure；
Fig. 7：Comparison study embodiment 2 carries out acipenser dabryanus fertilized eggs fertilized eggs phase histotomy figure.
Specific implementation mode
In order to make the object, technical scheme and advantages of the embodiment of the invention clearer, below in conjunction with the embodiment of the present invention
In attached drawing, technical scheme in the embodiment of the invention is clearly and completely described, it is clear that described embodiment is
A part of the embodiment of the present invention, instead of all the embodiments.Based on the embodiments of the present invention, those of ordinary skill in the art
The every other embodiment obtained without creative efforts, shall fall within the protection scope of the present invention.
Embodiment 1：
A kind of tissue section method of acipenser dabryanus fertilized eggs cleavage stage, comprises the steps of：
Material is fixed：The acipenser dabryanus ovum of development to fertilized eggs cleavage stage is put into Bouin ' the s fixers of 50 times of volumes
Fixed the ethyl alcohol for being 70% with volume fraction rinses repeatedly, is then stored in 70% ethyl alcohol for 24 hours, replaces every two days primary
70% alcohol continuously three times fully to wash away fixer, saves backup.
Dehydration：The fertilized eggs being stored in 70% ethyl alcohol are subjected to serial dehydration, 80% 2h of ethanol dehydration one time, 90% second
2h of dehydration of alcohols, 95% ethanol dehydration twice, each 2h, then twice with 100% ethanol dehydration, every time each dehydration 1h.
It is transparent：The fertilized eggs that dehydration is finished are placed in volume fraction terpinol：Absolute ethyl alcohol=1：In 1 transparent transition fluid
1h is handled, then handles 8h for the second time with 5h is handled in terpinol for the first time respectively.
Waxdip：Whole process carries out in 57 DEG C of constant temperature ovens.First merging terpinol and 57 DEG C of paraffin mix in equal volume
1h is handled in waxdip liquid, then passing through paraffin respectively again, processing 4h, paraffin handle 10h for the second time for the first time.
Embedding：By the sturgeon ovum conventional organization embedding method after the completion of waxdip, i.e., is embedded with carton, paraffin is made
Block.
Slice：Histotomy carries out conventional processing after slice, by the continuous paraffin section of slicer, by opening up piece, patch
Piece, roasting piece post-processing are spare.
Dyeing：Haematoxylin ＆ eosin (HE) dyes, and 37 DEG C of constant temperature oven drying of mounting, finally observation is taken pictures (such as Fig. 1-2).
Embodiment 2：
A kind of gastrul stage tissue section method of acipenser dabryanus fertilized eggs, comprises the steps of：
Material is fixed：The acipenser dabryanus fertilized eggs of development to different times are put into the formaldehyde fixer that mass fraction is 10%
For 24 hours, the ethyl alcohol for being 70% with volume fraction rinses repeatedly, is then stored in 70% ethyl alcohol for middle fixation, replaces every two days primary
70% alcohol continuously three times fully to wash away fixer, saves backup.
Dehydration：The fertilized eggs being stored in 70% ethyl alcohol are subjected to serial dehydration, 80% 2h of ethanol dehydration one time, 90% second
2h of dehydration of alcohols, 95% ethanol dehydration twice, each 2h, then twice with 100% ethanol dehydration, every time each dehydration 1h.
It is transparent：The fertilized eggs that dehydration is finished are placed in volume fraction terpinol：Absolute ethyl alcohol=1：In 1 transparent transition fluid
2h is handled, then handles 6h for the second time with 4h is handled in terpinol for the first time respectively.
Waxdip：Whole process carries out in 57 DEG C of constant temperature ovens.First merging terpinol and 57 DEG C of paraffin mix in equal volume
1h is handled in waxdip liquid, then passing through paraffin refined wax respectively again, processing 5h, paraffin refined wax handle 20h for the second time for the first time.
Embedding：By the sturgeon ovum conventional organization embedding method after the completion of waxdip, i.e., is embedded with carton, paraffin is made
Block.
Slice：Histotomy carries out conventional processing after slice, by the continuous paraffin section of slicer, by opening up piece, patch
Piece, roasting piece post-processing are spare.
Dyeing：Haematoxylin ＆ eosin (HE) dyes, and 37 DEG C of constant temperature oven drying of mounting, finally observation is taken pictures (such as Fig. 3).
Embodiment 3：
A kind of tissue section method of acipenser dabryanus fertilized eggs fertilized eggs phase, comprises the steps of：
Material is fixed：The acipenser dabryanus fertilized eggs of development to different times are put into Bouin ' the s fixers of 50 times of volumes
Fixed the ethyl alcohol for being 70% with volume fraction rinses repeatedly, is then stored in 70% ethyl alcohol for 24 hours, replaces every two days primary
70% alcohol continuously three times fully to wash away fixer, saves backup.
Dehydration：The fertilized eggs being stored in 70% ethyl alcohol are subjected to serial dehydration, 80% 2h of ethanol dehydration one time, 90% second
2h of dehydration of alcohols, 95% ethanol dehydration twice, each 1h, then twice with 100% ethanol dehydration, every time each dehydration 1h.
It is transparent：The fertilized eggs that dehydration is finished are placed in volume fraction terpinol：Absolute ethyl alcohol=1：In 1 transparent transition fluid
2h is handled, then handles 5h for the second time with 5h is handled in terpinol for the first time respectively.
Waxdip：Whole process carries out in 57 DEG C of constant temperature ovens.First merging terpinol and 57 DEG C of paraffin mix in equal volume
1h is handled in waxdip liquid, then passing through paraffin respectively again, processing 5h, paraffin handle 10h for the second time for the first time.
Embedding：By the sturgeon ovum conventional organization embedding method after the completion of waxdip, i.e., is embedded with carton, paraffin is made
Block.
Slice：Histotomy carries out conventional processing after slice, by the continuous paraffin section of slicer, by opening up piece, patch
Piece, roasting piece post-processing are spare.
Dyeing：Haematoxylin ＆ eosin (HE) dyes, and 37 DEG C of constant temperature oven drying of mounting, finally observation is taken pictures (such as Fig. 4).
Comparative example 1：
A kind of tissue section method of sturgeons fertilized eggs gastrul stage, comprises the steps of：
Step 1：The acipenser dabryanus fertilized eggs of development to different times are put into Bouin ' the s fixers of 50 times of volumes solid
Fixed the ethyl alcohol for being 70% with volume fraction rinses repeatedly, is then stored in 70% ethyl alcohol for 24 hours, replaces one time 70% every two days
Alcohol continuously three times fully to wash away fixer, saves backup.
Step 2：The fertilized eggs being stored in 70% ethyl alcohol are subjected to serial dehydration, 80% ethanol dehydration one time 2h, 90%
2h of ethanol dehydration, 95% ethanol dehydration twice, each 2h, then twice with 100% ethanol dehydration, every time each dehydration 1h.
Step 3：The fertilized eggs that dehydration is finished are placed in volume fraction terpinol：Absolute ethyl alcohol=1：1 transparent transition fluid
Then middle processing 0.5h handles 15min for the second time with 15min is handled in terpinol for the first time respectively.
Step 4：Whole process carries out in 57 DEG C of constant temperature ovens.It is first placed in terpinol and 57 DEG C of paraffin mixes in equal volume
Waxdip liquid in handle 1h, then pass through paraffin processing 2h for the first time respectively again, paraffin handles 2h for the second time.
Step 5：Paraffin embedding is carried out, paraffin mass is made.
Step 6：Histotomy carries out conventional processing, haematoxylin ＆ eosin (HE) dyeing, 37 DEG C of constant temperature of mounting after slice
Baking oven is dried, and finally observation is taken pictures (such as Fig. 5).
Comparative example 2：
A kind of tissue section method of sturgeons fertilized eggs fertilized eggs phase, comprises the steps of：
Step 1：The acipenser dabryanus fertilized eggs of development to different times are put into Bouin ' the s fixers of 50 times of volumes solid
Fixed the ethyl alcohol for being 70% with volume fraction rinses repeatedly, is then stored in 70% ethyl alcohol for 24 hours, replaces one time 70% every two days
Alcohol continuously three times fully to wash away fixer, saves backup.
Step 2：Egg membrane is poked on fertilized eggs surface with dissecting needle, is placed in culture dish, prepares 1% warm agar, slowly
Slowly culture dish is poured into, covers fertilized eggs, natural cooling is cut into agar block with scalpel.
Step 2：Agar block containing fertilized eggs is subjected to serial dehydration, 80% 2h of ethanol dehydration one time, 90% ethyl alcohol are de-
2h of water, 95% ethanol dehydration twice, each 2h, then twice with 100% ethanol dehydration, every time each dehydration 1h.
Step 3：The agar block that dehydration is finished is placed in dimethylbenzene：Absolute ethyl alcohol=1：15min in 1 transparent transition fluid,
Then 5min is handled for the second time with 10min is handled in terpinol for the first time respectively.
Step 4：Waxdip, whole process carry out in 57 DEG C of constant temperature ovens.First it is placed in the mixed liquid such as terpinol and 57 DEG C of paraffin
Middle 1h, then passing through paraffin respectively again, processing 1.5h, paraffin handle 1.5h for the second time for the first time.
Step 5：Paraffin embedding is carried out, paraffin mass is made.
Step 6：Histotomy carries out conventional processing, haematoxylin ＆ eosin (HE) dyeing, 37 DEG C of constant temperature of mounting after slice
Baking oven is dried, and finally observation is taken pictures (such as Fig. 6 and 7).
Using the fracture of slice egg membrane, Yolk disperse, egg membrane made from comparative example it can be seen from attached drawing 6-7
It is detached with Yolk, egg membrane fracture cannot reach requirement.
Claims (7)
1. a kind of tissue section method of acipenser dabryanus fertilized eggs, it is characterised in that：It fixed, be dehydrated by material, is transparent, waxdip, packet
It buries, be sliced and seven steps compositions of dyeing；The transparent step is after transparent liquid is handled, then passes through respectively at terpinol first time
Reason 4-5h and terpinol handle 5-7h for the second time, until the transparent state of sturgeon ovum；The dehydration total time is 10-20h；The leaching
The wax time is 16-30h；
The dehydration is the sturgeon ovum stored in 70% ethyl alcohol is directly immersed in 80% ethanol dehydration primary, time 2h, and
Pass sequentially through 90% ethanol dehydration 2h, 95% ethanol dehydration twice, each 1-2h, 100% ethanol dehydration twice,
Each 1-2h.
2. a kind of tissue section method of acipenser dabryanus fertilized eggs as described in claim 1, it is characterised in that：The material is fixed
Step is by fresh acipenser dabryanus fish-egg is put into Bouin ' the s fixers of 40-60 times of volume or mass fraction is 10% formaldehyde
It after fixing 20-30 hours in fixer, is rinsed, is then stored in the ethyl alcohol that volume fraction is 70% repeatedly with 70% ethyl alcohol,
It is 70% ethyl alcohol to replace volume fraction every two days, continuously three times after saved backup in 70% ethyl alcohol.
3. a kind of tissue section method of acipenser dabryanus fertilized eggs as claimed in claim 1 or 2, it is characterised in that：The material
Fixing step is that fresh acipenser dabryanus fish-egg is put into Bouin ' the s fixers of 50 times of volumes fixed or 10% formaldehyde to consolidate
Determine after fixing 24 hours in liquid, be stored in after being rinsed repeatedly with 70% ethyl alcohol in 70% ethyl alcohol, replaces 70% wine every two days
Essence continuously saves backup afterwards three times.
4. a kind of tissue section method of acipenser dabryanus fertilized eggs as described in claim 1, it is characterised in that：The transparent step
It is terpinol for first volume ratio will be placed in through dewatered sturgeon ovum：Absolute ethyl alcohol=1：1 transparent transition fluid or volume ratio
For terpinol：Dimethylbenzene：Absolute ethyl alcohol=4：1：1-2h is handled in 5 mixed transparent transition fluid, then passes through pine tar respectively again
Processing 4-5h and terpinol handle 5-7h to alcohol for the second time for the first time, until the transparent state of sturgeon ovum.
5. a kind of tissue section method of acipenser dabryanus fertilized eggs as described in claim 1, it is characterised in that：The waxdip step
1-2h is handled first to change to the sturgeon ovum after transparent processing in the waxdip liquid that terpinol mixes in equal volume with paraffin, is then divided again
Not Jing Guo paraffin for the first time processing 5-8h and paraffin handle 10-20h for the second time；The waxdip process in 57 DEG C of constant temperature ovens into
Row.
6. a kind of tissue section method of acipenser dabryanus fertilized eggs as described in claim 1, it is characterised in that：The embedding step
By the sturgeon ovum conventional organization embedding method after the completion of waxdip, i.e., to be embedded with carton, paraffin mass being made.
7. a kind of tissue section method of acipenser dabryanus fertilized eggs as described in claim 1, it is characterised in that：The slice and dye
Color step is by the continuous paraffin section of slicer, by opening up piece, patch, baking piece, by haematoxylin ＆ eosin (HE) dyeing, finally
Microexamination is taken pictures.
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| CN110361242B (en) * | 2019-08-14 | 2022-01-18 | 武汉赛维尔生物科技有限公司 | Fixing liquid for eyeball tissue and pretreatment method for preparing eyeball tissue slices |
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