CN102184670A - Hucho trout biological slice making technology - Google Patents
Hucho trout biological slice making technology Download PDFInfo
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- CN102184670A CN102184670A CN2011100282841A CN201110028284A CN102184670A CN 102184670 A CN102184670 A CN 102184670A CN 2011100282841 A CN2011100282841 A CN 2011100282841A CN 201110028284 A CN201110028284 A CN 201110028284A CN 102184670 A CN102184670 A CN 102184670A
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Abstract
The invention discloses a hucho trout biological slice making technology, and relates to a slice making technology. The technology of the invention solves the problems existing in the conventional slice making technology. The problems are that the slice is incomplete, the slice is not clear after slice sealing, and the agent is harmful. The making method includes the steps of: (1) fixing a hucho trout sample by employing the Bouin's solution, washing the sample by running water, and placing the sample into a first liquid, a second liquid, and terpineol in turn; (2) taking out the sample from the terpineol and placing into a mixed liquid which has equal volumes of terpineol and paraffin, and then placing the sample into the paraffin for embedding, slicing the sample after cooling; (3) carrying out the HE dyeing to the slice, and obtaining a finished product after sealing the slice by Euparal. The hucho trout glass sample obtained through the hucho trout biological slice making technology has the characteristics of fresh dyeing color, clear acid and alkaline cells, complete slice composition, clear image, and well-defined outline. The making technology of the invention saves many steps compared with the conventional making method, and reduces environmental pollution.
Description
Technical field
The present invention relates to a kind of tabletting technology.
Background technology
At present, the conventional organic section making technology that adopts all is with the organic solvent of benzene alcohols as clarifier and paraffin, some biomaterials meeting drawdown deformations through the effect of benzene alcohols, the sclerosis embrittlement can not be made into complete slice, thin piece, causes experiment to proceed, and benzene class material is to people's all toxic effect of blood forming organ, nervous system, digestive system, as prolonged and repeated contact benzene class, even nervous centralis there is the paralysis effect, also can causes environmental pollution simultaneously; Conventional organic section making technology is to adopt neutral gum as mountant, but has the problem that does not see Chu after the mounting.Existing conventional organic section making technology specific aim is poor, lacks the tabletting technology at single species.
Summary of the invention
The invention provides a kind of taimen organic section making technology, purpose is to have in order to solve existing conventional organic section making technology that film-making is imperfect, do not see Chu and the harmful problem of reagent after the mounting.
Taimen organic section making technology is carried out according to the following steps: one, the taimen sample adopts that BouinShi is liquid-solid to decide 2h, washes 10min with flowing water then, places first liquid, second liquid and each 30min of terpinol (Terpineol) more successively; Two, the taimen sample takes out from terpinol and is placed on 30～40min in terpinol and the isopyknic mixed liquor of paraffin, takes out to be placed on that 2h carries out embedding in the paraffin, cuts into slices after the cooling under the room temperature; Three, HE dyeing (hematoxylin eosin staining method) is carried out in section, adopt then Euparal (English to Chinese is: mounting excellent Ba Laer), promptly finish the taimen organic section making;
Wherein the taimen sample is the various tissues of taimen in the step 1; In the step 1 first liquid by volume umber form by 6 parts of absolute ethyl alcohols, 3 parts of chloroforms and 1 part of glacial acetic acid; In the step 1 second liquid by volume umber form by 6 parts of normal butyl alcohols, 3 parts of chloroforms and 1 part of glacial acetic acid.
The present invention adopts the clarifier terpinol (Terpineol) that has dehydration concurrently to replace absolute ethyl alcohol and dimethylbenzene to make paraffin section, and with Euparal (excellent Ba Laer) mounting, the benzene alcohols that has changed employing always is as organic solvent, neutral gum is as mountant, this has not only solved the difficult problem of organic section making technology, and more conventional method for making to have saved many steps poly-, reduced environmental pollution, obtained satisfied effect.
It is that it can be miscible with pure and mild ether that the present invention adopts the advantage of terpinol, is insoluble in water, also solubilized paraffin, excellent Ba Laer, and nontoxic.
The present invention adopts the advantage of Euparal to be: 1, biological specimen can directly take out mounting from the ethanol immobile liquid; 2, the general 24h of fast drying; 3, index of refraction be 1.483 than neutral gum (1.578) for low, relatively near the refraction coefficient (about 1.4) of animal and plant cells, therefore, many dyeing or the very shallow tissue of dyeing do not see in neutral gum, just can show with excellent Ba Laer mounting.
The taimen slide sample of taimen organic section making technology gained of the present invention, have dyeing fresh, the soda acid cell is clearly demarcated, film-making structural integrity, clear picture, the characteristics of sharp outline.
Description of drawings
Fig. 1 is the micrograph of the slide sample of the ovum of gained taimen maturation in the embodiment nine; Fig. 2 is the micrograph of the slide sample of gained taimen oral epithelium tissue in the embodiment ten; Fig. 3 is the micrograph of the slide sample of gained taimen pyloric caecum in the embodiment 11; Fig. 4 is the micrograph of the slide sample of gained taimen ovary in the embodiment 12.
Embodiment
Embodiment one: present embodiment taimen organic section making technology is carried out according to the following steps: one, the taimen sample adopts that BouinShi is liquid-solid to decide 2h, wash 10min with flowing water then, place first liquid, second liquid and each 30min of terpinol (Terpineol) more successively; Two, the taimen sample takes out from terpinol and is placed on 30～40min in terpinol and the isopyknic mixed liquor of paraffin, takes out to be placed on that 2h carries out embedding in the paraffin, cuts into slices after the cooling under the room temperature; Three, HE dyeing (hematoxylin eosin staining method) is carried out in section, adopt then Euparal (English to Chinese is: mounting excellent Ba Laer), promptly finish the taimen organic section making;
Wherein the taimen sample is the various tissues of taimen in the step 1; In the step 1 first liquid by volume umber form by 6 parts of absolute ethyl alcohols, 3 parts of chloroforms and 1 part of glacial acetic acid; In the step 1 second liquid by volume umber form by 6 parts of normal butyl alcohols, 3 parts of chloroforms and 1 part of glacial acetic acid.
Taimen sample after flowing water dashes in the present embodiment step 1 can place volumetric concentration to be that 70% ethanol is medium-term and long-term and preserve.
(English to Chinese is: excellent Ba Laer), be existing commercially available prod, can buy and obtain for terpinol in the present embodiment (Terpineol) and Euparal.
Embodiment two: what present embodiment and embodiment one were different is that BouinShi liquid is made up of the formaldehyde of 25ml, the picric acid saturated solution of 75ml and the glacial acetic acid of 5ml in the step 1.Other step and parameter are identical with embodiment one.
Embodiment three: what present embodiment and embodiment one were different is that step 1 places terpinol 30min, middlely must change terpinol once.Other step and parameter are identical with embodiment one.
Embodiment four: what present embodiment and embodiment one were different is that step 1 places paraffin 2h to carry out embedding, middlely must change paraffin once.Other step and parameter are identical with embodiment one.
Embodiment five: present embodiment and embodiment one are different is that the taimen sample takes out from terpinol and is placed on 30min in terpinol and the isopyknic mixed liquor of paraffin in the step 2.Other step and parameter are identical with embodiment one.
Embodiment six: present embodiment and embodiment one are different is that the taimen sample takes out from terpinol and is placed on 40min in terpinol and the isopyknic mixed liquor of paraffin in the step 2.Other step and parameter are identical with embodiment one.
Embodiment seven: present embodiment and embodiment one are different is that the taimen sample takes out from terpinol and is placed on 32～38min in terpinol and the isopyknic mixed liquor of paraffin in the step 2.Other step and parameter are identical with embodiment one.
Embodiment eight: present embodiment and embodiment one are different is that the taimen sample takes out from terpinol and is placed on 35min in terpinol and the isopyknic mixed liquor of paraffin in the step 2.Other step and parameter are identical with embodiment one.
Embodiment nine: present embodiment taimen organic section making technology is carried out according to the following steps: one, the taimen sample adopts that BouinShi is liquid-solid to decide 2h, washes 10min with flowing water then, places first liquid, second liquid and each 30min of terpinol more successively; Two, the taimen sample takes out from terpinol and is placed on 35min in terpinol and the isopyknic mixed liquor of paraffin, takes out to be placed on that 2h carries out embedding in the paraffin, cuts into slices after the cooling under the room temperature; Three, HE dyeing is carried out in section, adopts the Euparal mounting then, promptly finishes the taimen organic section making;
Wherein the taimen sample is the ovum of taimen maturation in the step 1; In the step 1 first liquid by volume umber form by 6 parts of absolute ethyl alcohols, 3 parts of chloroforms and 1 part of glacial acetic acid; In the step 1 second liquid by volume umber form by 6 parts of normal butyl alcohols, 3 parts of chloroforms and 1 part of glacial acetic acid.
The slide sample of the ovum of gained taimen maturation in the present embodiment, in Fig. 1, as seen, the soda acid cell of sample is clearly demarcated in the sample, film-making structural integrity, clear picture, sharp outline.
Embodiment ten: present embodiment taimen organic section making technology is carried out according to the following steps: one, the taimen sample adopts that BouinShi is liquid-solid to decide 2h, washes 10min with flowing water then, places first liquid, second liquid and each 30min of terpinol more successively; Two, the taimen sample takes out from terpinol and is placed on 30min in terpinol and the isopyknic mixed liquor of paraffin, takes out to be placed on that 2h carries out embedding in the paraffin, cuts into slices after the cooling under the room temperature; Three, HE dyeing is carried out in section, adopts the Euparal mounting then, promptly finishes the taimen organic section making;
Wherein the taimen sample is the oral epithelium tissue of taimen in the step 1; In the step 1 first liquid by volume umber form by 6 parts of absolute ethyl alcohols, 3 parts of chloroforms and 1 part of glacial acetic acid; In the step 1 second liquid by volume umber form by 6 parts of normal butyl alcohols, 3 parts of chloroforms and 1 part of glacial acetic acid.
The slide sample of gained taimen oral epithelium tissue in the present embodiment, in Fig. 2, as seen, the soda acid cell of sample is clearly demarcated in the sample, film-making structural integrity, clear picture, sharp outline.
Embodiment 11: present embodiment taimen organic section making technology is carried out according to the following steps: one, the taimen sample adopts that BouinShi is liquid-solid to decide 2h, washes 10min with flowing water then, places first liquid, second liquid and each 30min of terpinol more successively; Two, the taimen sample takes out from terpinol and is placed on 40min in terpinol and the isopyknic mixed liquor of paraffin, takes out to be placed on that 2h carries out embedding in the paraffin, cuts into slices after the cooling under the room temperature; Three, HE dyeing is carried out in section, adopts the Euparal mounting then, promptly finishes the taimen organic section making;
Wherein the taimen sample is the pyloric caecum of taimen in the step 1; In the step 1 first liquid by volume umber form by 6 parts of absolute ethyl alcohols, 3 parts of chloroforms and 1 part of glacial acetic acid; In the step 1 second liquid by volume umber form by 6 parts of normal butyl alcohols, 3 parts of chloroforms and 1 part of glacial acetic acid.
The slide sample of gained taimen pyloric caecum in the present embodiment, in Fig. 3, as seen, the soda acid cell of sample is clearly demarcated in the sample, film-making structural integrity, clear picture, sharp outline.
Embodiment 12: present embodiment taimen organic section making technology is carried out according to the following steps: one, the taimen sample adopts that BouinShi is liquid-solid to decide 2h, washes 10min with flowing water then, places first liquid, second liquid and each 30min of terpinol more successively; Two, the taimen sample takes out from terpinol and is placed on 30～40min in terpinol and the isopyknic mixed liquor of paraffin, takes out to be placed on that 2h carries out embedding in the paraffin, cuts into slices after the cooling under the room temperature; Three, HE dyeing is carried out in section, adopts the Euparal mounting then, promptly finishes the taimen organic section making;
Wherein the taimen sample is the ovary of taimen in the step 1; In the step 1 first liquid by volume umber form by 6 parts of absolute ethyl alcohols, 3 parts of chloroforms and 1 part of glacial acetic acid; In the step 1 second liquid by volume umber form by 6 parts of normal butyl alcohols, 3 parts of chloroforms and 1 part of glacial acetic acid.
The slide sample of gained taimen ovary in the present embodiment, in Fig. 4, as seen, the soda acid cell of sample is clearly demarcated in the sample, film-making structural integrity, clear picture, sharp outline.
Claims (6)
1. taimen organic section making technology, it is characterized in that taimen organic section making technology carries out according to the following steps: one, the taimen sample adopts the liquid-solid 2h of deciding of BouinShi, wash 10min with flowing water then, place first liquid, second liquid and each 30min of terpinol more successively; Two, the taimen sample takes out from terpinol and is placed on 30～40min in terpinol and the isopyknic mixed liquor of paraffin, takes out to be placed on that 2h carries out embedding in the paraffin, cuts into slices after the cooling under the room temperature; Three, HE dyeing is carried out in section, adopts the Euparal mounting then, promptly finishes the taimen organic section making;
Wherein the taimen sample is the various tissues of taimen in the step 1; In the step 1 first liquid by volume umber form by 6 parts of absolute ethyl alcohols, 3 parts of chloroforms and 1 part of glacial acetic acid; In the step 1 second liquid by volume umber form by 6 parts of normal butyl alcohols, 3 parts of chloroforms and 1 part of glacial acetic acid.
2. a kind of taimen organic section making technology according to claim 1 is characterized in that BouinShi liquid is made up of the formaldehyde of 25ml, the picric acid saturated solution of 75ml and the glacial acetic acid of 5ml in the step 1.
3. a kind of taimen organic section making technology according to claim 1 is characterized in that step 1 places terpinol 30min, and middle palpus is changed terpinol once.
4. a kind of taimen organic section making technology according to claim 1 is characterized in that step 1 places paraffin 2h to carry out embedding, and middle palpus is changed paraffin once.
5. a kind of taimen organic section making technology according to claim 1 is characterized in that the taimen sample takes out in the step 2 to be placed on 32～38min in terpinol and the isopyknic mixed liquor of paraffin from terpinol.
6. a kind of taimen organic section making technology according to claim 1 is characterized in that the taimen sample takes out in the step 2 to be placed on 35min in terpinol and the isopyknic mixed liquor of paraffin from terpinol.
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN2011100282841A CN102184670A (en) | 2011-01-26 | 2011-01-26 | Hucho trout biological slice making technology |
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN2011100282841A CN102184670A (en) | 2011-01-26 | 2011-01-26 | Hucho trout biological slice making technology |
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| CN102184670A true CN102184670A (en) | 2011-09-14 |
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| CN2011100282841A Pending CN102184670A (en) | 2011-01-26 | 2011-01-26 | Hucho trout biological slice making technology |
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Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102607930A (en) * | 2012-03-13 | 2012-07-25 | 中国水产科学研究院黑龙江水产研究所 | Fish germ cell orientated embedding technology |
| CN103076283A (en) * | 2013-01-17 | 2013-05-01 | 中国水产科学研究院黑龙江水产研究所 | Method for observing developmental process of salmonidae roes |
| CN103884556A (en) * | 2014-03-21 | 2014-06-25 | 海南大学 | Determination method for quality of frozen tilapia mossambica slices |
| CN105241698A (en) * | 2015-09-25 | 2016-01-13 | 四川农业大学 | Preparation method of Euchiloglanis kishinouyei Kimu-ra skin paraffin sections |
Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101871855A (en) * | 2009-04-27 | 2010-10-27 | 上海蓝盎电子科技发展有限公司 | Novel pathological section clarifier |
-
2011
- 2011-01-26 CN CN2011100282841A patent/CN102184670A/en active Pending
Patent Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101871855A (en) * | 2009-04-27 | 2010-10-27 | 上海蓝盎电子科技发展有限公司 | Novel pathological section clarifier |
Non-Patent Citations (2)
| Title |
|---|
| 《水产学杂志》 19960531 关海红 《鱼类组织蜡切片技术的改进》 第65-67页 1-6 第9卷, 第1期 * |
| 《水产学杂志》 20020531 关海红，曲秋芝 《利用松油醇对鲟鱼成熟卵子组织结构的观察》 第71-73页 1-6 第15卷, 第1期 * |
Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102607930A (en) * | 2012-03-13 | 2012-07-25 | 中国水产科学研究院黑龙江水产研究所 | Fish germ cell orientated embedding technology |
| CN103076283A (en) * | 2013-01-17 | 2013-05-01 | 中国水产科学研究院黑龙江水产研究所 | Method for observing developmental process of salmonidae roes |
| CN103884556A (en) * | 2014-03-21 | 2014-06-25 | 海南大学 | Determination method for quality of frozen tilapia mossambica slices |
| CN105241698A (en) * | 2015-09-25 | 2016-01-13 | 四川农业大学 | Preparation method of Euchiloglanis kishinouyei Kimu-ra skin paraffin sections |
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Application publication date: 20110914 |