CN109374373A - A kind of Apple paraffin section fast method for preparing - Google Patents
A kind of Apple paraffin section fast method for preparing Download PDFInfo
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- CN109374373A CN109374373A CN201811174149.6A CN201811174149A CN109374373A CN 109374373 A CN109374373 A CN 109374373A CN 201811174149 A CN201811174149 A CN 201811174149A CN 109374373 A CN109374373 A CN 109374373A
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- 239000012188 paraffin wax Substances 0.000 title claims abstract description 78
- 238000000034 method Methods 0.000 title claims abstract description 31
- 238000012545 processing Methods 0.000 claims abstract description 28
- 238000004519 manufacturing process Methods 0.000 claims abstract description 26
- 238000004043 dyeing Methods 0.000 claims abstract description 22
- 230000018044 dehydration Effects 0.000 claims abstract description 16
- 238000006297 dehydration reaction Methods 0.000 claims abstract description 16
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims abstract description 16
- 238000010186 staining Methods 0.000 claims abstract description 7
- 241000220225 Malus Species 0.000 claims description 85
- CTQNGGLPUBDAKN-UHFFFAOYSA-N O-Xylene Chemical compound CC1=CC=CC=C1C CTQNGGLPUBDAKN-UHFFFAOYSA-N 0.000 claims description 39
- 239000001993 wax Substances 0.000 claims description 20
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims description 18
- 235000019441 ethanol Nutrition 0.000 claims description 8
- 235000021016 apples Nutrition 0.000 claims description 5
- 239000012153 distilled water Substances 0.000 claims description 5
- 230000007935 neutral effect Effects 0.000 claims description 5
- 238000001035 drying Methods 0.000 claims description 4
- IINOLFPQEZQVMB-UHFFFAOYSA-N ethanol;1,2-xylene Chemical compound CCO.CC1=CC=CC=C1C IINOLFPQEZQVMB-UHFFFAOYSA-N 0.000 claims description 4
- 238000011010 flushing procedure Methods 0.000 claims description 4
- 239000007788 liquid Substances 0.000 claims description 3
- 229950003937 tolonium Drugs 0.000 claims description 3
- HNONEKILPDHFOL-UHFFFAOYSA-M tolonium chloride Chemical compound [Cl-].C1=C(C)C(N)=CC2=[S+]C3=CC(N(C)C)=CC=C3N=C21 HNONEKILPDHFOL-UHFFFAOYSA-M 0.000 claims description 3
- WRMNZCZEMHIOCP-UHFFFAOYSA-N 2-phenylethanol Chemical compound OCCC1=CC=CC=C1 WRMNZCZEMHIOCP-UHFFFAOYSA-N 0.000 claims description 2
- 239000005357 flat glass Substances 0.000 claims 1
- 239000008096 xylene Substances 0.000 claims 1
- 235000013399 edible fruits Nutrition 0.000 abstract description 23
- 238000011160 research Methods 0.000 abstract description 9
- 238000001514 detection method Methods 0.000 abstract description 7
- 238000002360 preparation method Methods 0.000 abstract description 3
- 230000014509 gene expression Effects 0.000 abstract description 2
- 238000007901 in situ hybridization Methods 0.000 abstract description 2
- 238000011017 operating method Methods 0.000 abstract description 2
- 238000003892 spreading Methods 0.000 abstract 1
- 210000004027 cell Anatomy 0.000 description 15
- 239000000975 dye Substances 0.000 description 8
- 230000000694 effects Effects 0.000 description 8
- WSFSSNUMVMOOMR-UHFFFAOYSA-N Formaldehyde Chemical compound O=C WSFSSNUMVMOOMR-UHFFFAOYSA-N 0.000 description 6
- 238000011161 development Methods 0.000 description 5
- 239000011521 glass Substances 0.000 description 5
- QTBSBXVTEAMEQO-UHFFFAOYSA-N Acetic acid Chemical compound CC(O)=O QTBSBXVTEAMEQO-UHFFFAOYSA-N 0.000 description 4
- 238000001574 biopsy Methods 0.000 description 4
- 210000002421 cell wall Anatomy 0.000 description 3
- 230000002380 cytological effect Effects 0.000 description 3
- 241000196324 Embryophyta Species 0.000 description 2
- URLKBWYHVLBVBO-UHFFFAOYSA-N Para-Xylene Chemical group CC1=CC=C(C)C=C1 URLKBWYHVLBVBO-UHFFFAOYSA-N 0.000 description 2
- YXFVVABEGXRONW-UHFFFAOYSA-N Toluene Natural products CC1=CC=CC=C1 YXFVVABEGXRONW-UHFFFAOYSA-N 0.000 description 2
- 229960000583 acetic acid Drugs 0.000 description 2
- 239000003086 colorant Substances 0.000 description 2
- 239000012362 glacial acetic acid Substances 0.000 description 2
- 238000010827 pathological analysis Methods 0.000 description 2
- 108090000623 proteins and genes Proteins 0.000 description 2
- 238000007711 solidification Methods 0.000 description 2
- 230000008023 solidification Effects 0.000 description 2
- 241000693079 Maloideae Species 0.000 description 1
- 206010054949 Metaplasia Diseases 0.000 description 1
- 230000002411 adverse Effects 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 230000001413 cellular effect Effects 0.000 description 1
- 238000005520 cutting process Methods 0.000 description 1
- 230000008034 disappearance Effects 0.000 description 1
- 238000009826 distribution Methods 0.000 description 1
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- 238000011156 evaluation Methods 0.000 description 1
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- 238000010438 heat treatment Methods 0.000 description 1
- 238000011065 in-situ storage Methods 0.000 description 1
- 238000002386 leaching Methods 0.000 description 1
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- 230000035772 mutation Effects 0.000 description 1
- 238000005457 optimization Methods 0.000 description 1
- 238000004321 preservation Methods 0.000 description 1
- 230000001737 promoting effect Effects 0.000 description 1
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Classifications
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N1/00—Sampling; Preparing specimens for investigation
- G01N1/28—Preparing specimens for investigation including physical details of (bio-)chemical methods covered elsewhere, e.g. G01N33/50, C12Q
- G01N1/30—Staining; Impregnating ; Fixation; Dehydration; Multistep processes for preparing samples of tissue, cell or nucleic acid material and the like for analysis
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- Life Sciences & Earth Sciences (AREA)
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- General Health & Medical Sciences (AREA)
- General Physics & Mathematics (AREA)
- Immunology (AREA)
- Pathology (AREA)
- Agricultural Chemicals And Associated Chemicals (AREA)
- Investigating Or Analysing Biological Materials (AREA)
Abstract
The present invention provides a kind of Apple paraffin section fast method for preparing, it is fixed including sample, sample dehydration, transparent processing, sample embedding, sample slice, spread out piece processing and tissue staining, by spreading out piece treatment process, reduce the surface tension of water and paraffin in paraffin section, cell tissue boundary is set to generate certain water phase thin layer, highlight cell tissue structure, it is easy to dye on boundary, production method to simplify fruit paraffin section in the prior art, the production of Apple paraffin section is set to no longer need to the dewaxing after being sliced, rehydration, and the dehydration after dyeing, the operating procedures such as transparent, reduce the microsection manufacture time at present no more than 2 days by original 1 week or so time, significantly shorten fabrication cycle, improve producing efficiency, chipping qualities can also be fully ensured that simultaneously, reduce preparation cost , the scientific research for fruit tree cultivation technical field cytology and in situ hybridization gene expression detection etc. provides technical support.
Description
Technical field
The present invention relates to a kind of Apple paraffin section production methods, and in particular to a kind of Apple paraffin section is fast
Fast production method.
Background technique
Paraffin section production is to carry out a basic experiment technological means of plant tissue cell's analysis, and traditional paraffin is cut
Piece method includes all multi-steps such as fixation, dehydration, transparent, waxdip, slice, dewaxing, rehydration, dyeing, dehydration, transparent, operation
Process complexity is cumbersome, and preparation cost is higher, and fabrication cycle is longer, generally needs one week or so from starting to be fixed to complete
Time, biggish workload and longer Production Time, to plant tissue cell learn and in-situ techniques analysis gene tissue and
Spatial and temporal distributions research in cell produces great adverse effect, also carries out related scientific research work to researcher and brings
Very big puzzlement.Apple belongs to rosaceae Maloideae, is perennial deciduous fruit tree, and apple variety is resourceful, fruit size
For the fruit size difference that difference, especially bud mutation generate in addition to through phenotypic evaluation, cytological evidence analyzes the original that difference is formed
Because also particularly significant, histocyte analysis is carried out to Apple at present and uses more or traditional paraffin section method.
But since Apple water content is more, required dewatering time is longer, during microsection manufacture, often will appear fruit
Metaplasia, dehydration is not thorough enough, it is chipping and other issues to be sliced, in sample dewaxing process, since the disappearance of wax makes to glue
Attached effect dies down, and slice easily falls off from glass slide, causes microsection manufacture to fail, more important is due to traditional paraffin section
Production method complex steps, period are longer, it is difficult to quickly observation and analysis are made to the difference of fruit cell, to correlation
The development of aspect scientific research, teaching and pathological analysis work produces serious restriction.At present for fruit especially Apples
The research of real slice fast method for preparing is also rarely reported, and therefore, research is established the efficient fruit paraffin of fast and stable and cut
Piece production method has become an important topic for promoting fruit tree cultivation technical field research.
Summary of the invention
It is an object of the present invention to provide a kind of Apple paraffin section fast method for preparing, simplify the Apple
The manufacturing process of paraffin section shortens fabrication cycle, improves chipping qualities, reduces preparation cost.
A kind of Apple paraffin section fast method for preparing, comprises the steps of:
Step 1, sample are fixed: being cut Apple sample to be tested, quickly fixed at least 24 hours using fixer, make institute
The protein stated in Apple sample is sufficiently denaturalized, and maintains the stabilization of cell tissue；
Wherein, Apple development cuts the entire slice of position 0.5cm thickness among fruit, apple when fruit is smaller early period
When fruit transverse diameter or vertical diameter are bordering on coverslip size, 1cm × 1cm × 0.5cm fixed dimension slice is cut, the fixer is preferred
Use FAA(50% ethyl alcohol: formaldehyde: glacial acetic acid=90: 5: 5).
Step 2, sample dehydration: the Apple sample that step 1 fixes is carried out step by step with different concentration ethanol
Dehydration, dehydration concentration gradient are followed successively by 50% → 70% → 85% → 95% → 100% → 100%, and every grade of dewatering time is 1h, makes institute
Stating Apple sample, to reach tissue dewatering thorough, keeps cellular morphology complete；
Wherein, in order to keep the Apple sample dehydration more uniform thoroughly, preferably in dehydration step by step, by the apple
Fruits sample is put in plastic centrifuge tube, while the centrifuge tube being placed on shaking table and is shaken.
Step 3, transparent processing: the Apple sample dimethylbenzene dewatered to step 2 and diformazan benzyl carbinol are molten
Liquid carries out transparent processing, and concentration for the treatment of gradient is followed successively by dimethylbenzene ethanol solution (Er Jia Ben ︰ ethyl alcohol=1 ︰ 1) → dimethylbenzene → bis-
Toluene, every grade of processing time is 1h, keeps the Apple sample tissue substantially transparent.
Step 4, sample embedding: the Apple sample of the step 3 after transparent is carried out through dimethylbenzene and paraffin 65
The waxdip of DEG C mixed liquor to pure wax twice is handled, it may be assumed that dimethylbenzene and paraffin mixed liquor (Er Jia Ben ︰ paraffin=1 ︰ 1) 65 DEG C of leachings overnight
Wax → 65 DEG C, which are uncapped, makes dimethylbenzene volatilization 2h → paraffin refined wax waxdip 2h → paraffin refined wax waxdip 2h → tissue embedding machine embedding, makes described
Apple sample waxdip is complete；
Wherein, sample embeds preferred purified paraffin, free from foreign meter, without filtering after wax melting；Before waxdip preferably by dimethylbenzene and
Paraffin to prevent dimethylbenzene in room temperature from may cause the solidification of paraffin, guarantees that the dimethylbenzene and paraffin are mixed in 65 DEG C of water-bath 1h
It closes liquid and more comes into full contact with the Apple sample, be put in 65 DEG C of oven overnight waxdips；Meanwhile last time paraffin refined wax waxdip
It is preferably placed in embedding machine wax pan and carries out, to prevent from may cause hollow wax caused by paraffin quickly solidifies too far because of migration distance
The phenomenon that block.
Step 5, sample slice: the Apple sample for completing embedding to step 4 carries out slicing treatment, after repairing wax
It is sliced using slicer, slice thickness is 10~15 μm, this wax of Apple band is made to keep connection intact；
Wherein, period determination is educated when the specific slice thickness of the Apple sample is sent out according to the Apple, before development
Phase cell arrangement is close, selects lower thickness, and development later stage space between cells increases, and selects thicker degree.
Step 6, booth piece processing: booth piece processing is carried out to the Apple sample that step 5 completes slice, by the apple
It is open and flat to be placed in the fixed position of glass slide after fruits slice is using 40 DEG C of water-bath exhibition pieces of water-bath, using booth piece machine 38~45
DEG C stand booth piece 2h, so that the Apple sample is reached state easy to dye and be adhered on glass slide.
Wherein, it is reduced due to water in heating state lower surface tension, same paraffin in a heated state also can by surface tension
It reduces, in the case where the water and loose paraffin surface tension, it is thin that cell tissue boundary can generate certain water phase
Layer, highlights cell tissue structure, is easy to dye on boundary；
Step 7, tissue staining: to step 6 booth piece, treated that the Apple sample carries out tissue staining, uses concentration 1%
Toluidine blue the Apple sample is dyed, dye 2~5min；
Wherein, the specific dyeing time of the Apple sample is determined according to the Apple developmental stage, develops early period
Cell arrangement is close, selects longer dyeing time, and development later stage space between cells increases, and selects shorter dyeing time.
Step 8, mounting processing: mounting processing is carried out to the Apple sample after step 7 dyeing, by color card
Using distilled water flushing it is clean after, after drying using a small amount of neutral gum carry out mounting processing, complete the Apple paraffin
The production of slice.
Wherein, after using distilled water flushing clean color card, before carrying out mounting to sample, one is preferably carried out again
Secondary booth piece processing, it may be assumed that the color card rinsed well is placed on the piece machine of booth, booth piece is stood at a temperature of 38~45 DEG C
2h carries out mounting processing after drying.
It is cut by a kind of Apple paraffin that Apple paraffin section fast method for preparing completes
Piece, neutral gum mounting directly can carry out cytological observation using aobvious emblem mirror immediately after the completion, if not being eager to be observed,
It can be observed at any time after neutral gum solidification, it can using the Apple paraffin section that technical solution of the present invention makes
Preservation 1 year or more time.
Fruit paraffin section is all to be mainly based upon water using the method that rehydration dyes again after dewaxing in the prior art
Property coloring agent is incompatible with paraffin, easily causes cell that can not dye or dye non-uniform problem；A kind of apple of the present invention
Fruits paraffin section fast method for preparing is spread out at piece after dyeing again using the direct staining after the piece processing of booth after slice
The method of reason, the method are to pass through certain temperature and time conditions using the skin effect between embedding paraffin and cell wall
Under water-bath exhibition piece and booth piece process, promote to generate certain physical chemistry and electrochemical effect between hydrone and cell wall,
Make to generate a continuous water phase thin layer on cell wall, water dyeing agent can dye the water phase thin layer；Meanwhile it dyeing
The booth of optimization again piece processing afterwards can be such that coloring agent further spreads in the water phase thin layer, play sufficiently dyeing and homochromatic
Effect；The Apple paraffin section made of technical solution of the present invention, through micro- sem observation, as shown in Figure 1 to Figure 3, carefully
Born of the same parents show that completely stitch clarity is very high, can meet the needs of scientific research, teaching and pathological analysis well.
The beneficial effects of the present invention are: proposing a kind of Apple paraffin section fast method for preparing, manufacturing process is reduced
Time-consuming, demanding operating procedure for dewaxing, rehydration after middle slice and the dehydration after dyeing, transparent etc., overcomes existing skill
The drawbacks of fruit paraffin section making step is cumbersome in art, excessive cycle, the production of Apple paraffin section is time-consuming
Within original reduction in 1 week as little as 2 days, it is quick, stable, high to meet current fruit tree cultivation technical field fruit texture
Imitate the needs of detection；Meanwhile also solve Apple paraffin section dyeing after dewatering time it is longer cause biopsy tissues deformation,
The problems such as chipping, and adhesion effect decrease causes paraffin section easily to fall off from glass slide after dewaxing, improve apple
The chipping qualities and producing efficiency of fruit paraffin section, reduce and manufacture cost, are fruit tree cultivation technical field cytological observation
And the research of in situ hybridization gene expression detection etc. provides good technical support.
Detailed description of the invention
12 days after Fig. 1, full blossom, " Long Feng " Apple paraffin section that slice thickness is 10 μm, dyeing time is 5min
Heterogeneous microstructure photo.
86 days after Fig. 2, full blossom, " Long Feng " Apple paraffin section that slice thickness is 13 μm, dyeing time is 3min
Heterogeneous microstructure photo.
120 days after Fig. 3, full blossom, " Long Feng " Apple paraffin section that slice thickness is 15 μm, dyeing time is 2min
Heterogeneous microstructure photo.
Specific embodiment
Combined with specific embodiments below and attached drawing, the claimed technical solution of the present invention is further described.
Embodiment 1
A kind of fast method for preparing of Apple paraffin section, selection are grown on southeast part of Heilongjiang Province In Mudanjiang District full blossom
12 days " Long Feng " Apples are detection sample afterwards, set slice thickness as 10 μm, dyeing time 5min, production method by
Following steps composition:
Step 1: cutting the Apple detection sample of certain volume, cut sample is quickly fixed on FAA(50% second
Alcohol: formaldehyde: glacial acetic acid=90: 5: 5) in, the set time is for 24 hours；
The Apple sample fixed is dehydrated by step 2 step by step with different concentration ethanol, dehydration concentration gradient according to
Secondary is 50% → 70% → 85% → 95% → 100% → 100%, and every grade of dewatering time is 1h, during the dehydration process, by the apple
Fruit sample is put in the plastic centrifuge tube of 50ml, while the centrifuge tube being placed on shaking table and is shaken；
Step 3 carries out transparent processing to the dewatered Apple sample with dimethylbenzene and dimethylbenzene ethanol solution, processing
Concentration gradient is followed successively by dimethylbenzene ethanol solution (Er Jia Ben ︰ ethyl alcohol=1 ︰ 1) → dimethylbenzene → dimethylbenzene, and every grade of processing time is equal
For 1h；
Step 4 selects 60 DEG C of Leica paraffin to embed the Apple sample after transparent, first paraxylene and
Paraffin is in 65 DEG C of water-bath 1h, then through dimethylbenzene and paraffin mixed liquor (Er Jia Ben ︰ paraffin=1 ︰ 1) 65 DEG C of overnight waxdip → 65 DEG C
Uncapping makes dimethylbenzene volatilization 2h → paraffin refined wax waxdip 2h → paraffin refined wax waxdip 2h → tissue embedding machine embedding, and embedding sample is made；
Wherein, last time paraffin refined wax waxdip is placed in embedding machine wax pan and carries out；
Step 5 cuts the Apple sample of embedding after repairing wax using LeicaRM2255 cycle type slicer
Piece；
Step 6 carries out booth piece processing to the Apple sample paraffin section using LeicaHI1220 water-bath booth piece machine, makes
With water-bath 40 DEG C through water-bath open up piece after, it is open and flat be placed on glass slide LeicaHI1220 booth piece machine on keep 40 DEG C standing
Spread out piece 2h；
Step 7 carries out tissue staining using the toluidine blue of concentration 1% to through booth piece treated the Apple sample, it
The color card is rinsed well using distilled water afterwards；
The color card is placed on the piece machine of the booth LeicaHI1220 by step 8, is stood booth piece 2h again at a temperature of 40 DEG C, is dried in the air
Mounting is carried out using a small amount of neutral gum after dry.
The Apple paraffin section of the present embodiment production is used Olympus BX50F-3 to show emblem mirror and directly observes card
Bright, as shown in Figure 1, biopsy tissues transparent effect is excellent, dyeing clarity is high, and cell boundaries are complete, and tissue image is clear.
Embodiment 2
A kind of fast method for preparing of Apple paraffin section, selection are grown on southeast part of Heilongjiang Province In Mudanjiang District full blossom
86 days " Long Feng " Apples are detection sample afterwards, sets slice thickness as 13 μm, dyeing time 3min, production method and
Embodiment 1 is identical.
The Apple paraffin section of the present embodiment production is used Olympus BX50F-3 to show emblem mirror and directly observes card
Bright, as shown in Fig. 2, biopsy tissues transparent effect is excellent, dyeing clarity is high, and cell boundaries are complete, and tissue image is clear.
Embodiment 3
A kind of fast method for preparing of Apple paraffin section, selection are grown on southeast part of Heilongjiang Province In Mudanjiang District full blossom
120 days " Long Feng " Apples are detection sample afterwards, set slice thickness as 15 μm, dyeing time 2min, production method
It is same as Example 1.
The Apple paraffin section of the present embodiment production is used Olympus BX50F-3 to show emblem mirror and directly observes card
Bright, as shown in figure 3, biopsy tissues transparent effect is excellent, dyeing clarity is high, and cell boundaries are complete, and tissue image is clear.
Claims (7)
1. a kind of Apple paraffin section fast method for preparing, which is characterized in that comprise the steps of:
Step 1, sample are fixed: being cut Apple sample to be tested, quickly fixed at least for 24 hours using fixer；
Step 2, sample dehydration: being dehydrated the Apple sample that step 1 fixes with different concentration ethanol step by step,
Dehydration concentration gradient is followed successively by 50% → 70% → 85% → 95% → 100% → 100%, and every grade of dewatering time is 1h；
Step 3, transparent processing: Apple sample diformazan benzyl carbinol dewatered to step 2 and xylene solution into
Row transparent processing, concentration for the treatment of gradient are followed successively by dimethylbenzene ethanol solution (Er Jia Ben ︰ ethyl alcohol=1 ︰ 1) → dimethylbenzene → dimethylbenzene,
Every grade of processing time is 1h；
Step 4, sample embedding: the Apple sample of the step 3 after transparent is carried out mixed at 65 DEG C through dimethylbenzene and paraffin
The waxdip for closing liquid to pure wax twice is handled, it may be assumed that and 65 DEG C of overnight waxdips of dimethylbenzene and paraffin mixed liquor (Er Jia Ben ︰ paraffin=1 ︰ 1) →
Uncapping for 65 DEG C makes dimethylbenzene volatilization 2h → paraffin refined wax waxdip 2h → paraffin refined wax waxdip 2h → tissue embedding machine embedding；
Step 5, sample slice: the Apple sample for completing embedding to step 4 carries out slicing treatment, uses after repairing wax
Slicer is sliced, and slice thickness is 10~15 μm；
Step 6, booth piece processing: booth piece processing is carried out to the Apple sample that step 5 completes slice, by the Apples
After real slice is using 40 DEG C of water-bath exhibition pieces of water-bath, the open and flat glass slide that is placed in fixes position, quiet at 38~45 DEG C using booth piece machine
Set booth piece 2h；
Step 7, tissue staining: to step 6 booth piece, treated that the Apple sample carries out tissue staining, uses concentration 1%
Toluidine blue the Apple sample is dyed, dye 2~5min；
Step 8, mounting processing: mounting processing is carried out to the Apple sample after step 7 dyeing, color card is used
After distilled water flushing is clean, mounting processing is carried out using a small amount of neutral gum after drying, completes the Apple paraffin section
Production.
2. a kind of Apple paraffin section fast method for preparing as described in claim 1, it is characterised in that: the step 2
In, in dehydration step by step, the Apple sample is put in plastic centrifuge tube, while the centrifuge tube being placed in and is shaken
It is shaken on bed.
3. a kind of Apple paraffin section fast method for preparing as described in claim 1, it is characterised in that: the step 4
In, it the use of the paraffin is 60 DEG C of Leica paraffin, by dimethylbenzene and the paraffin in 65 DEG C of water-bath 1h before waxdip.
4. a kind of Apple paraffin section fast method for preparing as described in claim 1, it is characterised in that: the step 6
In booth piece temperature be 40 DEG C.
5. such as a kind of described in any item Apple paraffin section fast method for preparing of Claims 1-4, it is characterised in that:
In the step 7, after using distilled water flushing clean color card, before carrying out mounting to sample, carried out at the piece of booth again
Reason, it may be assumed that the color card rinsed well is placed on the piece machine of booth, booth piece 2h is stood at a temperature of 38~45 DEG C, after drying
Carry out mounting processing.
6. a kind of Apple paraffin section fast method for preparing as claimed in claim 5, it is characterised in that: the step 7
The booth piece temperature for carrying out booth piece processing after middle dyeing again is 40 DEG C.
7. a kind of Apple paraffin section fast method for preparing as described in claim 1, it is characterised in that: make in step 1
It is for 24 hours with fixer rapid immobilisation times.
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| CN110132690A (en) * | 2019-06-05 | 2019-08-16 | 河南城建学院 | A kind of toluidine blue rapid dyeing method of cerebral tissue frozen section |
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| CN112798391A (en) * | 2021-01-11 | 2021-05-14 | 四川农业大学 | Method for preparing bulbil paraffin embedding slice |
| CN114383906A (en) * | 2021-12-22 | 2022-04-22 | 中国农业科学院棉花研究所 | Preparation method of plant root system paraffin section |
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