CN101790975B - Manufacturing method of daphnia microslide specimen - Google Patents
Manufacturing method of daphnia microslide specimen Download PDFInfo
- Publication number
- CN101790975B CN101790975B CN 201010136398 CN201010136398A CN101790975B CN 101790975 B CN101790975 B CN 101790975B CN 201010136398 CN201010136398 CN 201010136398 CN 201010136398 A CN201010136398 A CN 201010136398A CN 101790975 B CN101790975 B CN 101790975B
- Authority
- CN
- China
- Prior art keywords
- daphnia
- ethanol
- microslide
- specimen
- dyeing
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Expired - Fee Related
Links
Landscapes
- Investigating Or Analysing Biological Materials (AREA)
- Sampling And Sample Adjustment (AREA)
Abstract
The invention provides a manufacturing method of a daphnia microslide specimen, which comprises the following steps: firstly fixing the daphnia with a fixative, simple-staining or double-staining, color separating, and stepwise dehydrating with ethanol; then stepwise displacing the ethanol in the dehydrated daphnia with acetone; and finally sucking out the daphnia which is subject to the displacement with acetone, dripping on a glass slide, and dripping prepolymerized polymethacrylate to seal the glass slide to obtain a daphnia microslide specimen. The daphnia microslide specimen manufactured by the method enables the cell borderlines of the daphnia to be obvious, the intestinal tract, ovary and other structures to be observed more clearly, the transparency is increased, and the drying time is shortened.
Description
Technical field
The present invention relates to biological sample and make, be specifically related to a kind of preparation method of permanent daphnia microslide specimen.
Background technology
The whole method of tableting of present microscope slide sample used has: 1, temporary transient and semipermanent method of tableting comprises glycerine method, glycerin jelly method and lactic acid-phynol method; 2, permanent whole method of tableting comprises Venice turpentine method, tert-butyl alcohol natural gum method and dioxane natural gum method etc.; 3, to amber preparation of specimen method of larger insect etc.But for the water Magna materials such as Daphnia magna, sword water Magna sample as a whole, can not need dewater, directly use the glycerin cement mounting, operating process is simple like this, but has some problems, and glycerin cement drying time is long and easily slide, and causes the displacement of sample.Simultaneously health is transparent for water Magna, observes directly the interior of health by microscopes, but generally observes unclearly, needs dimethylbenzene to carry out carrying out mounting after transparent again.Mounting often adopts canada balsam or resinene, although the technical parameters such as their light transmittance, ultraviolet blocking-up, refraction meet the requirement of biological microscope slide sample sealing, and the fine structure indistinguishables such as the enteron aisle in its body, ovary, oculomotor muscle.
Summary of the invention
The object of the present invention is to provide a kind of form that can keep sample, can clearly observe again the preparation method of the daphnia microslide specimen of fine structure in body.
The preparation method of a kind of daphnia microslide specimen provided by the invention comprises the steps:
1) with water Magna with fixer fix, singly dye, color separation, ethanol dewater step by step;
2) will dewater after ethanol in water Magna body replace step by step with acetone, the ethanol that is about in water Magna body is replaced through 35% acetone → 50% acetone → 75% acetone → 85% acetone → 95% acetone → anhydrous propanone → anhydrous propanone → anhydrous propanone, each gradient 15-20 minute;
3) the water Magna after the acetone displacement is dropped on slide with the suction pipe sucking-off, drip polymethacrylates and carry out mounting; The proportioning of described polymethacrylates is:
Because polymethacrylates change in volume before and after polymerization is larger, cell structure there is certain damage, need to carry out prepolymerization.After before use raw material being mixed in above-mentioned ratio, the limit heating edge is shaken in 70-80 ℃ of water-bath, makes it aggregate into the thick of glycerine sample, and bubble reduces gradually, and after cooling, viscosity can slightly increase, and namely can be used for mounting.Seal and be placed in 50-60 ℃ of baking oven 24-36 hour after sheet and get final product.
Described is fixedly that the water Magna that will live was placed in fixer 1-2 hour.Described fixer is Bouin ' s fixer, 70% ethanolic solution or 4% neutral formalin solution.
Described singly dying for carmine dyeing dyeed 1 day.
Described color separation is that the water Magna after dyeing is carried out color separation with hydrochloric acid-alcohol.Also can be at ethanol step by step in dehydration, drip 1-2 and drip hydrochloric acid and get final product from 35%-75% ethanol.
Described ethanol dewaters step by step, be water Magna after fixing through 35% ethanol → 45% ethanol → 55% ethanol → 75% ethanol → 85% ethanol → 95% ethanol → absolute ethyl alcohol (absolute ethyl alcohol 3 times) (each gradient 15-20 minute), the moisture in water Magna body is replaced by ethanol gradually.
Described step 1) can be: with water Magna with fixer fix, the rather dyeing of chrome alum-cyanogen, color separation, orange G dyeing, ethanol dewaters step by step; Described chrome alum-cyanogen rather dyes, and dyeing time is 1-4 hour, and the peaceful proportioning of chrome alum-cyanogen is: the peaceful 0.1-0.4g of cyanogen, chrome alum 3-5g, 45% glacial acetic acid 100ml; Orange G dyeing, with the 95% saturated orange G dye liquor of ethanol configuration, dyeing time is 1-2 hour.
In fixing, dyeing, dehydration, replacement process, the vacuum available pump vacuumizes, and can add the speed of fast response, the shortening time.
The preparation method of microscope slide sample of the present invention also can be used for the microscope slide sample of fish-egg, microplankton etc. and makes.
The present invention compares with existing technology has following characteristics: (1), water Magna integral body is singly dyed (carmine) and redyed (the peaceful orange G of chrome alum cyanogen), organ, tissue and cell structure are obvious.(2), dewatering step by step by ethanol to cause the contraction of tissue and cell, therefore can not cause water Magna distortion.(3) sample is after acetone displacement, refraction index changing, and transparency improves, and interior is clear.(4) adopt polymethacrylates to carry out mounting, the transparency of sample is increased, dry time shorten.
Embodiment
Embodiment 1
Fixing: as the water Magna that lives to be placed in 70% ethanol water and to anaesthetize and fix.
Singly dye: carmine was contaminated 1 day.
Color separation: hydrochloric acid-alcohol color separation namely drips 1-2 and drips hydrochloric acid, until till falling without dyestuff in 70% ethanol.
Ethanol dewaters step by step: 85% ethanol → 95% ethanol → absolute ethyl alcohol dehydration (absolute ethyl alcohol 3 times) (each gradient 15 minutes).
Displacement: the ethanol after dewatering in water Magna body is through 35% acetone → 50% acetone → 75% acetone → 85% acetone → 95% acetone → anhydrous propanone displacement (anhydrous propanone 3 times) (each gradient 15 minutes).
Mounting: the water Magna in acetone directly is put on clean slide with the suction pipe sucking-off, a little after the evaporation, drip 1,2 prepolymerized methacrylate of process, carefully cover upper cover glass, avoid producing bubble, seal and be placed in 60 ℃ of baking ovens 36 hours after sheet, take out and namely make daphnia microslide specimen.
The proportioning of polymethacrylates is:
Embodiment 2
Fixing: that the water Magna that lives was placed in Bouin ' s fixer 1 hour.
Dyeing: first carry out chrome alum-cyanogen and rather dye, dyeed 3 hours.
Color separation: hydrochloric acid-alcohol color separation.
Ethanol dewaters step by step: through 35% ethanol → 45% ethanol → 55% ethanol → 75% ethanol → 85% ethanol → 95% ethanol → absolute ethyl alcohol → absolute ethyl alcohol → absolute ethyl alcohol dehydration, each gradient 20 minutes.
Redye: when being dewatered to 95% ethanol, prepare saturated orange G ethanolic solution, contaminated 1 hour.
Displacement: the ethanol after dewatering in water Magna body is through 35% acetone → 50% acetone → 75% acetone → 85% acetone → 95% acetone → anhydrous propanone → anhydrous propanone → anhydrous propanone displacement, each gradient 15 minutes.
Mounting: the water Magna in acetone directly is put on clean slide with the suction pipe sucking-off, a little after the evaporation, drip 1,2 prepolymerized methacrylate of process, carefully cover upper cover glass, avoid producing bubble, seal and be placed in 50 ℃ of baking ovens 24 hours after sheet, take out and namely make daphnia microslide specimen.
The proportioning of polymethacrylates is:
Embodiment 3
The water Magna that lives is placed in 4% neutral formalin solution is fixed, all the other make daphnia microslide specimen with embodiment 2.
Respectively at the optical microphotograph Microscopic observation, the cell boundary line of water Magna is obvious with embodiment 1,2 and 3 daphnia microslide specimens that make, and the structure observation such as enteron aisle, ovary get more clear, and transparency is good.
Claims (3)
1. the preparation method of a daphnia microslide specimen, is characterized in that, comprises the steps:
1) with water Magna with fixer fix, singly dye, color separation, ethanol dewater step by step;
2) will dewater after ethanol in water Magna body replace step by step with acetone;
3) the water Magna after the acetone displacement is dropped on slide with the suction pipe sucking-off, drip polymethacrylates and carry out mounting, obtain daphnia microslide specimen; The proportioning of described polymethacrylates is:
Described polymethacrylates is that after mixing by described polymethacrylates material rate before use, the limit heating edge is shaken in 70-80 ℃ of water-bath, makes it aggregate into the thick of glycerine sample;
Described fixer is Bouin ' s fixer, 70% ethanolic solution or 4% neutral formalin solution.
2. the preparation method of a kind of daphnia microslide specimen as claimed in claim 1, is characterized in that, described singly dying is carmine dyeing.
3. the preparation method of a kind of daphnia microslide specimen as claimed in claim 1 or 2, is characterized in that, described step 1) be: with water Magna with fixer fix, the rather dyeing of chrome alum-cyanogen, color separation, orange G dyeing, ethanol dewaters step by step;
Described chrome alum-cyanogen rather dyes, and dyeing time is 1-4 hour, and the peaceful proportioning of chrome alum-cyanogen is: the peaceful 0.1-0.4g of cyanogen, chrome alum 3-5g, 45% glacial acetic acid 100ml; Described orange G dyeing, with the 95% saturated orange G dye liquor of ethanol configuration, dyeing time is 1-2 hour.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN 201010136398 CN101790975B (en) | 2010-03-26 | 2010-03-26 | Manufacturing method of daphnia microslide specimen |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN 201010136398 CN101790975B (en) | 2010-03-26 | 2010-03-26 | Manufacturing method of daphnia microslide specimen |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN101790975A CN101790975A (en) | 2010-08-04 |
| CN101790975B true CN101790975B (en) | 2013-06-05 |
Family
ID=42584075
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN 201010136398 Expired - Fee Related CN101790975B (en) | 2010-03-26 | 2010-03-26 | Manufacturing method of daphnia microslide specimen |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN101790975B (en) |
Families Citing this family (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103293155B (en) * | 2013-05-17 | 2016-12-28 | 苏州大学 | A kind of freeze transparent tissue observation procedure |
| CN103868774A (en) * | 2014-03-26 | 2014-06-18 | 青岛大学 | Method for manufacturing parasitic ovum permanent slide specimens |
| CN104082275B (en) * | 2014-07-02 | 2016-03-23 | 青岛农业大学 | A kind of aquatic livestock embryo microscope slide sample manufacturing technology |
| CN104048868A (en) * | 2014-07-02 | 2014-09-17 | 青岛农业大学 | Zooplankton microscopic slide specimen preparation method |
| CN105076108B (en) * | 2015-07-07 | 2018-05-01 | 安徽师范大学 | The short-term fixed store method and long-term fixed store method of wheel animalcule sample |
| CN109827832A (en) * | 2019-01-17 | 2019-05-31 | 河南师范大学 | The production method of branch angular motion object Permanent slide |
| CN109749634B (en) * | 2019-02-25 | 2021-07-23 | 暨南大学 | Rapid encapsulating tablet based on photocuring and preparation method and application thereof |
Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1918983A (en) * | 2006-09-12 | 2007-02-28 | 山西大学 | Production method of insect specimen |
| CN101571626A (en) * | 2009-06-01 | 2009-11-04 | 高丽英 | Method for transparently sealing biological microscope slide sample |
| CN101650273A (en) * | 2009-07-30 | 2010-02-17 | 浙江万里学院 | Paraffin wax slicing method of ocean shellfish oocyte |
-
2010
- 2010-03-26 CN CN 201010136398 patent/CN101790975B/en not_active Expired - Fee Related
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1918983A (en) * | 2006-09-12 | 2007-02-28 | 山西大学 | Production method of insect specimen |
| CN101571626A (en) * | 2009-06-01 | 2009-11-04 | 高丽英 | Method for transparently sealing biological microscope slide sample |
| CN101650273A (en) * | 2009-07-30 | 2010-02-17 | 浙江万里学院 | Paraffin wax slicing method of ocean shellfish oocyte |
Also Published As
| Publication number | Publication date |
|---|---|
| CN101790975A (en) | 2010-08-04 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN101790975B (en) | Manufacturing method of daphnia microslide specimen | |
| CN102116711B (en) | Manufacturing method of paraffin sections of zostera marina embryo | |
| CN107167350B (en) | Preparation method of paraffin section of eggplant rhizome tissue | |
| CN107490511B (en) | A kind of haematoxylin dyeing liquid and HE colouring method | |
| CN102967493A (en) | Rapid paraffin sectioning method for plant tissue | |
| CN102279121B (en) | Green making technology of animal tissue paraffin section | |
| CN104977202A (en) | Preparation method of transmission electron microscope sample of paraffin-embedded section tissue | |
| CN103940648A (en) | Preparation method for gill tissue paraffin section | |
| CN105885070A (en) | Preparation method of regenerated silk fibroin membrane | |
| CN104987862A (en) | Carbon dot with continuous golgi apparatus target imaging capability and preparation method thereof | |
| Susaki et al. | Perspective: extending the utility of three-dimensional organoids by tissue clearing technologies | |
| CN109776819A (en) | A kind of bletilla polysaccharide-carboxymethyl chitosan composite hydrogel and its preparation | |
| CN105928752B (en) | A kind of insect imago paraffin section colouring method | |
| CN103940647A (en) | Manufacture method of continuous paraffin sections for gonad of pelodiscus sinensis in embryonic period and application of manufacture method in sex determination | |
| CN104483178A (en) | Preparation method of chromosomes of adult epinephelus akaara | |
| CN105241698A (en) | Preparation method of Euchiloglanis kishinouyei Kimu-ra skin paraffin sections | |
| CN108680418A (en) | A kind of rapid fluorescence colouring method of crop in cruciferae pollen | |
| CN105784705A (en) | Forensic medicine diatom test method | |
| CN107202720B (en) | A kind of paraffin section method of pomegranate seed | |
| CN111829859A (en) | Efficient transparent dyeing and three-dimensional imaging method for poplar seeds | |
| CN109859304B (en) | Three-dimensional printing technology for establishing three-dimensional structure digital model outside cornea limbal tissue | |
| CN110487830A (en) | A kind of rapid prototyping method of the tender plant of children for scanning electron microscopic observation | |
| CN104048868A (en) | Zooplankton microscopic slide specimen preparation method | |
| Jocic et al. | Fabrication of user-friendly and biomimetic 1, 1′-carbonyldiimidazole cross-linked gelatin/agar microfluidic devices | |
| CN108546681A (en) | Cell carves surface and its application again |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| C14 | Grant of patent or utility model | ||
| GR01 | Patent grant | ||
| CF01 | Termination of patent right due to non-payment of annual fee |
Granted publication date: 20130605 Termination date: 20180326 |
|
| CF01 | Termination of patent right due to non-payment of annual fee |