CN110426259A - Polyethylene glycol is used for the application of animal tissue sections grease dyeing - Google Patents
Polyethylene glycol is used for the application of animal tissue sections grease dyeing Download PDFInfo
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- CN110426259A CN110426259A CN201910749883.9A CN201910749883A CN110426259A CN 110426259 A CN110426259 A CN 110426259A CN 201910749883 A CN201910749883 A CN 201910749883A CN 110426259 A CN110426259 A CN 110426259A
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- polyethylene glycol
- dyeing
- animal tissue
- grease
- carbowax
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- 229920001223 polyethylene glycol Polymers 0.000 title claims abstract description 71
- 239000002202 Polyethylene glycol Substances 0.000 title claims abstract description 31
- 238000004043 dyeing Methods 0.000 title claims abstract description 29
- 241001465754 Metazoa Species 0.000 title claims abstract description 28
- 239000004519 grease Substances 0.000 title claims abstract description 19
- 210000001519 tissue Anatomy 0.000 claims abstract description 37
- 238000000034 method Methods 0.000 claims abstract description 22
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims abstract description 19
- NPGIHFRTRXVWOY-UHFFFAOYSA-N Oil red O Chemical compound Cc1ccc(C)c(c1)N=Nc1cc(C)c(cc1C)N=Nc1c(O)ccc2ccccc12 NPGIHFRTRXVWOY-UHFFFAOYSA-N 0.000 claims abstract description 14
- 239000012188 paraffin wax Substances 0.000 claims abstract description 11
- 238000005406 washing Methods 0.000 claims abstract description 7
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims description 23
- 235000019441 ethanol Nutrition 0.000 claims description 13
- 210000005228 liver tissue Anatomy 0.000 claims description 11
- PEDCQBHIVMGVHV-UHFFFAOYSA-N Glycerine Chemical compound OCC(O)CO PEDCQBHIVMGVHV-UHFFFAOYSA-N 0.000 claims description 10
- VEXZGXHMUGYJMC-UHFFFAOYSA-N Hydrochloric acid Chemical compound Cl VEXZGXHMUGYJMC-UHFFFAOYSA-N 0.000 claims description 10
- 239000012153 distilled water Substances 0.000 claims description 9
- WZUVPPKBWHMQCE-XJKSGUPXSA-N (+)-haematoxylin Chemical compound C12=CC(O)=C(O)C=C2C[C@]2(O)[C@H]1C1=CC=C(O)C(O)=C1OC2 WZUVPPKBWHMQCE-XJKSGUPXSA-N 0.000 claims description 5
- VHUUQVKOLVNVRT-UHFFFAOYSA-N Ammonium hydroxide Chemical compound [NH4+].[OH-] VHUUQVKOLVNVRT-UHFFFAOYSA-N 0.000 claims description 5
- 239000001828 Gelatine Substances 0.000 claims description 5
- WZUVPPKBWHMQCE-UHFFFAOYSA-N Haematoxylin Natural products C12=CC(O)=C(O)C=C2CC2(O)C1C1=CC=C(O)C(O)=C1OC2 WZUVPPKBWHMQCE-UHFFFAOYSA-N 0.000 claims description 5
- 241001062009 Indigofera Species 0.000 claims description 5
- 239000000908 ammonium hydroxide Substances 0.000 claims description 5
- 229920000159 gelatin Polymers 0.000 claims description 5
- 235000019322 gelatine Nutrition 0.000 claims description 5
- 235000011187 glycerol Nutrition 0.000 claims description 5
- 238000007711 solidification Methods 0.000 claims description 4
- 230000008023 solidification Effects 0.000 claims description 4
- 238000001816 cooling Methods 0.000 claims description 2
- 239000000203 mixture Substances 0.000 claims description 2
- 150000002632 lipids Chemical class 0.000 abstract description 10
- 230000018044 dehydration Effects 0.000 abstract description 9
- 238000006297 dehydration reaction Methods 0.000 abstract description 9
- 239000003960 organic solvent Substances 0.000 abstract description 5
- 230000008520 organization Effects 0.000 abstract description 5
- 238000002360 preparation method Methods 0.000 abstract description 5
- 210000000805 cytoplasm Anatomy 0.000 abstract description 3
- 238000005516 engineering process Methods 0.000 abstract description 3
- 230000008595 infiltration Effects 0.000 abstract description 3
- 238000001764 infiltration Methods 0.000 abstract description 3
- 230000015572 biosynthetic process Effects 0.000 abstract description 2
- 239000013078 crystal Substances 0.000 abstract description 2
- 230000003118 histopathologic effect Effects 0.000 abstract description 2
- 238000003475 lamination Methods 0.000 abstract description 2
- 238000004519 manufacturing process Methods 0.000 abstract description 2
- 239000012024 dehydrating agents Substances 0.000 abstract 1
- 241000699666 Mus <mouse, genus> Species 0.000 description 21
- 210000004185 liver Anatomy 0.000 description 12
- 231100000240 steatosis hepatitis Toxicity 0.000 description 12
- 208000004930 Fatty Liver Diseases 0.000 description 10
- 206010019708 Hepatic steatosis Diseases 0.000 description 10
- 208000010706 fatty liver disease Diseases 0.000 description 10
- CTQNGGLPUBDAKN-UHFFFAOYSA-N O-Xylene Chemical compound CC1=CC=CC=C1C CTQNGGLPUBDAKN-UHFFFAOYSA-N 0.000 description 6
- 239000000975 dye Substances 0.000 description 5
- 210000004027 cell Anatomy 0.000 description 4
- LYCAIKOWRPUZTN-UHFFFAOYSA-N Ethylene glycol Chemical compound OCCO LYCAIKOWRPUZTN-UHFFFAOYSA-N 0.000 description 3
- 230000003902 lesion Effects 0.000 description 3
- 206010002091 Anaesthesia Diseases 0.000 description 2
- 102000004190 Enzymes Human genes 0.000 description 2
- 108090000790 Enzymes Proteins 0.000 description 2
- 230000037005 anaesthesia Effects 0.000 description 2
- 239000003795 chemical substances by application Substances 0.000 description 2
- 230000000052 comparative effect Effects 0.000 description 2
- 230000008602 contraction Effects 0.000 description 2
- 238000007710 freezing Methods 0.000 description 2
- 230000008014 freezing Effects 0.000 description 2
- 230000007935 neutral effect Effects 0.000 description 2
- 230000001575 pathological effect Effects 0.000 description 2
- 239000007787 solid Substances 0.000 description 2
- 230000007863 steatosis Effects 0.000 description 2
- 210000003934 vacuole Anatomy 0.000 description 2
- 201000001320 Atherosclerosis Diseases 0.000 description 1
- 229920002527 Glycogen Polymers 0.000 description 1
- 208000000501 Lipidoses Diseases 0.000 description 1
- 206010024585 Lipidosis Diseases 0.000 description 1
- 241000699670 Mus sp. Species 0.000 description 1
- 229930006000 Sucrose Natural products 0.000 description 1
- CZMRCDWAGMRECN-UGDNZRGBSA-N Sucrose Chemical compound O[C@H]1[C@H](O)[C@@H](CO)O[C@@]1(CO)O[C@@H]1[C@H](O)[C@@H](O)[C@H](O)[C@@H](CO)O1 CZMRCDWAGMRECN-UGDNZRGBSA-N 0.000 description 1
- 210000001789 adipocyte Anatomy 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 150000001875 compounds Chemical class 0.000 description 1
- 230000006378 damage Effects 0.000 description 1
- 230000007812 deficiency Effects 0.000 description 1
- 230000007850 degeneration Effects 0.000 description 1
- 238000001514 detection method Methods 0.000 description 1
- 238000003745 diagnosis Methods 0.000 description 1
- 230000004069 differentiation Effects 0.000 description 1
- 235000013399 edible fruits Nutrition 0.000 description 1
- 230000000694 effects Effects 0.000 description 1
- 244000144992 flock Species 0.000 description 1
- 239000012530 fluid Substances 0.000 description 1
- 239000011521 glass Substances 0.000 description 1
- 229940096919 glycogen Drugs 0.000 description 1
- 210000002216 heart Anatomy 0.000 description 1
- 238000009396 hybridization Methods 0.000 description 1
- JEIPFZHSYJVQDO-UHFFFAOYSA-N iron(III) oxide Inorganic materials O=[Fe]O[Fe]=O JEIPFZHSYJVQDO-UHFFFAOYSA-N 0.000 description 1
- 210000003734 kidney Anatomy 0.000 description 1
- 231100001231 less toxic Toxicity 0.000 description 1
- 239000007788 liquid Substances 0.000 description 1
- 239000000463 material Substances 0.000 description 1
- 239000000126 substance Substances 0.000 description 1
- 239000005720 sucrose Substances 0.000 description 1
- 229940045860 white wax Drugs 0.000 description 1
Classifications
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N1/00—Sampling; Preparing specimens for investigation
- G01N1/02—Devices for withdrawing samples
- G01N1/04—Devices for withdrawing samples in the solid state, e.g. by cutting
- G01N1/06—Devices for withdrawing samples in the solid state, e.g. by cutting providing a thin slice, e.g. microtome
-
- 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
- G01N1/286—Preparing specimens for investigation including physical details of (bio-)chemical methods covered elsewhere, e.g. G01N33/50, C12Q involving mechanical work, e.g. chopping, disintegrating, compacting, homogenising
-
- 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
-
- 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
- G01N1/286—Preparing specimens for investigation including physical details of (bio-)chemical methods covered elsewhere, e.g. G01N33/50, C12Q involving mechanical work, e.g. chopping, disintegrating, compacting, homogenising
- G01N2001/2873—Cutting or cleaving
Abstract
The invention belongs to histopathologic slide's manufacture technology fields, and in particular to polyethylene glycol is used for the application of animal tissue sections grease dyeing.Utilize the outstanding water soluble nature of polyethylene glycol, using polyethylene glycol as dehydrating agent, embed the infiltration for realizing medium, embedding forming simultaneously during the dehydration process, the organization embedding block of formation has the sShape features of paraffin mass, organization embedding block can remove embedding medium through washing simultaneously, without using organic solvent processing, the problem of organic solvent processing causes lipid to be lost is avoided.The histotomy of the method for the invention preparation avoids frozen section blocked up the problem of leading to tissue lamination with a thickness of 3~5 μm.The histotomy of the method for the invention preparation is after oil red O stain, it is permeated because polyethylene glycol is dehydrated, make most of fat drips are intact to be stored in cytoplasm, the problem of fat drips are displaced in frozen section is avoided, also avoids causing institutional framework incomplete because low temperature leads to the problem of ice crystal in frozen section.
Description
Technical field
The invention belongs to histopathologic slide's manufacture technology fields, and in particular to polyethylene glycol is for animal tissue sections oil
The application of rouge dyeing.
Background technique
Lipid is the general name of neutral fat, lipoid and its derivative, is primarily referred to as neutral fat.Under normal circumstances, grease removal
Fat is extracellular, other generally lose into the cell or only a small amount of fat drips.When lesion occurs, it will appear fat drips or fat drips in cell
It increased significantly, when especially steatosis occurs for the organa parenchymatosums such as the heart, liver, kidney, will appear vacuole not of uniform size in endochylema, need
Belong to steatosis or hydropic degeneration with fat stains identification vacuole or glycogen stores.In addition, when atherosclerosis, it is interior
Lipidosis under chrotoplast can show lipid with fat stains.
Lipid is soluble in organic solvent, cannot carry out diagnosis detection with paraffin section.Because paraffin section is in film-making, dyeing
It needs to carry out dewaxing treatment by organic solvents such as alcohol, dimethylbenzene in the process, lipid etc. can be removed.Therefore paraffin section can not
Pathological examination for lipid.
In daily pathological examination and scientific research, fat stains is frequently with frozen section film-making, oil red O stain.Frozen section
Preparation is that will organize after OCT embedding medium embeds forming, and tissue block is cut into 5~15 μ with freezing microtome under -20 DEG C of environment
Then the slice of m carries out oil red O stain.Blocked up due to being sliced in dyeing course, dyeing and when affixed slice, are easy to produce folding
It is folded, influence chipping qualities.Thermal expansion and contraction will lead to fat drips and flock together, and make fat cell in " empty balloon-shaped ", dyeing knot
Fruit is negative change, and is sliced and cannot be saved for a long time, need to be observed, be had some limitations in time.
Carbowax also known as polyethylene glycol are a kind of water-soluble high-molecular compounds, are had a series of from as low as intermediate molecular weight
Product.The physical aspect of carbowax can be from colorless and transparent viscous fluid (molecular weight 200~700) to white wax semisolid (molecule
Amount 1000~2000) up to hard waxy solid (molecular weight 3000~20000).Carbowax is completely soluble, and can and it is very much
Substance mixes, and has good stability and surface-active, less toxic and nonirritant.
Summary of the invention
The present invention is in view of the deficiencies of the prior art, and it is an object of the present invention to provide a kind of polyethylene glycol is used for animal tissue sections grease
The application of dyeing.
For achieving the above object, the technical scheme adopted by the invention is as follows:
Polyethylene glycol is used for the application of animal tissue sections grease dyeing.
A method of the dyeing of animal tissue sections grease being carried out using polyethylene glycol, is included the following steps:
(1) animal tissue's sample is taken, after fixer is fixed, flowing water is rinsed, blotting paper suck dry moisture；
(2) animal tissue's sample is placed in progress waxdip processing in carbowax I；
(3) animal tissue's sample is placed in progress waxdip processing in carbowax II again；
(4) then animal's liver tissue sample is placed in tissue embedding box, the carbowax II of enough fusings is added, it is naturally cold
But,
It after carbowax solidification forming, is sliced with paraffin slicing machine, normal-temperature distilled water opens up piece after slice, bakes piece；
(5) slice is slightly washed with distilled water without dewaxing, directly enters haematoxylin dyeing 2min；It slightly washes, 0.5% hydrochloric acid
Differentiation rapidly, 0.5% ammonium hydroxide returns indigo plant after washing；Then it is gently washed with 70% ethyl alcohol the slice several seconds, oil red O stain 10min, 70%
Ethyl alcohol breaks up rapidly, dries in air, glycerin gelatine mounting.
In above scheme, carbowax I is the polyethylene glycol of molecular weight 1000~2000.
In above scheme, carbowax II is that molecular weight 1500 is gathered with the polyethylene glycol of molecular weight 1000 as what 3:1~1:3 was mixed
Ethylene glycol mixture.
In above scheme, the condition of step (2) and step (3) the waxdip processing are as follows: be placed in 55~65 DEG C of incubators
Manage 1~3h.
In above scheme, step (3) the waxdip number of processing is 3~5 times.
In above scheme, the condition of step (4) the roasting piece are as follows: 2~4h of piece is baked in 37~42 DEG C of incubators.
In above scheme, animal tissue's embedded section described in step (4) with a thickness of 3~5 μm.
In the present invention, carbowax embedding has the Some features of frozen section and paraffin embedding:
It (1) can be solid at room temperature since carbowax is liquid at high temperature from " hydropexis phase " direct embedded section,
It can be dissolved in water by different proportion, can be directly immersed in histocyte；
(2) without dewaxing through dehydration of alcohol, dimethylbenzene after tissue is fixed, the carbowax of gradient molecular weight can directly be used
Dehydration embedding, avoids the fatsolvents such as alcohol, dimethylbenzene from utmostly saving lipid to the destruction of lipid；
(3) due to reducing the programs such as dehydration, transparent, tissue block contraction distortion can be avoided as far as possible, is saved in histocyte
Activity, the antigenicity of enzyme are completely suitable for the technologies such as enzyme histochemistry and molecule hybridization；
(4) it can be sliced with common paraffin slicing machine.
Beneficial effects of the present invention: (1) using polyethylene glycol outstanding water soluble nature, using polyethylene glycol as dehydration
Agent, embeds the infiltration for realizing medium, embedding forming simultaneously during the dehydration process, and the organization embedding block of formation has the outer of paraffin mass
Shape feature, while organization embedding block can remove embedding medium through washing, without using organic solvent processing, avoid organic molten
The problem of agent processing causes lipid to be lost.(2) histotomy of the method for the invention preparation avoids ice with a thickness of 3~5 μm
Freeze and is sliced blocked up the problem of leading to tissue lamination.(3) the method for the invention preparation histotomy after oil red O stain, because
Polyethylene glycol dehydration infiltration, makes most of fat drips are intact to be stored in cytoplasm, avoids fat drips in frozen section and be displaced
The problem of, it also avoids causing institutional framework incomplete because low temperature leads to the problem of ice crystal in frozen section.
Detailed description of the invention
Fig. 1 is normal mouse liver carbowax embedded section oil red O stain, photo under 200 times of mirrors.
Fig. 2 is fatty liver mouse liver carbowax embedded section oil red O stain, photo under 200 times of mirrors.
Fig. 3 is fatty liver mouse liver frozen section oil red O stain, photo under 200 times of mirrors.
Specific embodiment
For a better understanding of the present invention, below with reference to the embodiment content that the present invention is furture elucidated, but it is of the invention
Content is not limited solely to the following examples.
Embodiment 1
A method of mouse liver animal tissue sections grease is carried out using polyethylene glycol and is dyed, and is included the following steps:
(1) Normal healthy mice liver tissue sample is taken, about 0.5cm × 0.5cm × 0.5cm, after fixer is fixed,
Flowing water rinses, blotting paper suck dry moisture；
(2) mouse liver tissue sample is placed in carbowax I (polyethylene glycol that molecular weight is 1000), in 60 DEG C of incubators
Waxdip handles 60min；
(3) mouse liver tissue sample is placed in carbowax II again (carbowax of molecular weight 1500 and molecular weight 1000 is mixed by 1:2
Close) in, waxdip handles 60min in 60 DEG C of incubators；
(4) mouse liver tissue sample is placed in carbowax II again (carbowax of molecular weight 1500 and molecular weight 1000 is mixed by 1:1
Close) in, waxdip handles 60min in 60 DEG C of incubators；
(5) mouse liver tissue sample is placed in carbowax II again (carbowax of molecular weight 1500 and molecular weight 1000 is mixed by 2:1
Close) in, waxdip handles 60min in 60 DEG C of incubators；
(6) then mouse liver tissue sample is placed in tissue embedding box, the carbowax II (molecular weight of enough fusings is added
1500 are mixed with the carbowax of molecular weight 1000 by 2:1), natural cooling, after carbowax solidification forming, with paraffin section machine-cut
Piece, slice with a thickness of 5 μm, normal-temperature distilled water opens up piece after slice, and piece 2h is baked in 37 DEG C of incubators；
(7) slice is slightly washed with distilled water without dewaxing, directly enters haematoxylin dyeing 2min；It then slightly washes, 0.5%
Hydrochloric acid breaks up rapidly, and 0.5% ammonium hydroxide returns indigo plant after washing；70% ethyl alcohol is gently washed the slice several seconds, saturation oil red O dye liquor dyeing
10min；70% ethyl alcohol breaks up rapidly, dries in air, glycerin gelatine mounting.
The present embodiment is prepared into the animal tissue sections after grease obtained dyeing and is placed in microscopically observation, as a result such as Fig. 1
Shown, there are a small amount of fat drips in normal mouse liver, fat drips are rounded, and core week, accurate positioning are distributed in cytoplasm.
Embodiment 2
A method of fatty liver mouse liver slice grease is carried out using polyethylene glycol and is dyed, and is included the following steps:
(1) Models of Fatty Liver mouse takes liver tissue sample after anesthesia is put to death immediately, and about 0.5cm × 0.5cm ×
0.5cm, after fixer is fixed, flowing water is rinsed, blotting paper suck dry moisture；
(2) mouse fatty liver is placed in carbowax I (polyethylene glycol that molecular weight is 1000), in 55 DEG C of incubators at waxdip
Manage 90min；
(3) again by mouse fatty liver be placed in carbowax II (molecular weight 1500 mixed with the carbowax of molecular weight 1000 by 1:2 and
At) in, waxdip handles 120min in 55 DEG C of incubators；
(4) again by mouse fatty liver be placed in carbowax II (molecular weight 1500 mixed with the carbowax of molecular weight 1000 by 2:1 and
At) in, waxdip handles 120min in 55 DEG C of incubators；
(5) again by mouse fatty liver be placed in carbowax II (molecular weight 1500 mixed with the carbowax of molecular weight 1000 by 2:1 and
At) in, waxdip handles 120min in 55 DEG C of incubators；
(6) then mouse liver tissue sample is placed in tissue embedding box, the carbowax II of enough fusings is added, it is naturally cold
But, after carbowax solidification forming, be sliced with paraffin slicing machine, slice with a thickness of 4 μm, normal-temperature distilled water opens up piece after slice, in
Piece 2h is baked in 40 DEG C of incubators；
(7) slice is slightly washed with distilled water without dewaxing, directly enters haematoxylin dyeing 2min；It then slightly washes, 0.5%
Hydrochloric acid breaks up rapidly, and 0.5% ammonium hydroxide returns indigo plant after washing；70% ethyl alcohol is gently washed the slice several seconds, saturation oil red O dye liquor dyeing
10min；70% ethyl alcohol breaks up rapidly, dries in air, glycerin gelatine mounting.
The present embodiment is prepared into the animal tissue sections after grease obtained dyeing and is placed in microscopically observation, as a result such as Fig. 2
Fat lesion occurs for the liver of shown Models of Fatty Liver mouse, has the fat drips largely coloured by oil red, is positioned in endochylema, fat drips
It is not displaced, nucleus shows clear.
Comparative example
Included the following steps: in the prior art using the method for frozen section oil red O stain
(1) Models of Fatty Liver mouse takes liver to take liver tissue sample immediately after anesthesia is put to death, and about 0.5cm ×
0.5cm × 0.5cm, after fixer is fixed, flowing water is rinsed, blotting paper suck dry moisture.
(2) mouse liver is placed in 15~30% graded sucrose solutions and carries out 24~36h of dehydration, until tissue sinks to pipe
Bottom represents dehydration and completes；
(3) dewatered liver organization is embedded with OCT embedding medium, and freezing microtome slice, is cut by 8~10 μm of slice thickness
Piece is attached directly on anticreep glass slide, is saved in -20 DEG C；
(4) before dyeing, frozen section is slightly washed in room temperature rewarming 10min, distilled water, directly enters haematoxylin dyeing 2min；With
It slightly washes afterwards, 0.5% hydrochloric acid breaks up rapidly, and 0.5% ammonium hydroxide returns indigo plant after washing；
(5) 70% ethyl alcohol are gently washed the slice several seconds, and saturation oil red O dye liquor dyes 10min；70% ethyl alcohol breaks up rapidly, empty
It is dried in gas, glycerin gelatine mounting.
Fig. 3 is fat lesion mouse liver frozen section oil red O stain as a result, visible small fat drips shape is not true to type in figure,
Big fat drips obviously shift, position inaccurate, and fat drips are linked to be sheet and float on above tissue, cause nucleus very unobvious.
The comparative example that compares Fig. 3 is prepared after gained slice dyes using the method for the present invention (see Fig. 1 and Fig. 2), and lesser fat drips are in typical case
Graininess, big fat drips are respectively positioned on around cell within a cell core, and accurate positioning, nucleus is not covered by fat drips, and display is clear.
To sum up, it is believed that the carbowax embedded section flaking method that the present invention illustrates is better than the locating and displaying of lipid material in tissue
Existing frozen section flaking method.
Claims (8)
1. the application that polyethylene glycol is used for the dyeing of animal tissue sections grease.
2. a kind of method for carrying out the dyeing of animal tissue sections grease using polyethylene glycol, which comprises the steps of:
(1) animal tissue's sample is taken, after fixer is fixed, flowing water is rinsed, blotting paper suck dry moisture；
(2) animal tissue's sample is placed in progress waxdip processing in carbowax I；
(3) animal tissue's sample is placed in progress waxdip processing in carbowax II again；
(4) then animal's liver tissue sample is placed in tissue embedding box, is added the carbowax II of enough fusings, natural cooling,
It after carbowax solidification forming, is sliced with paraffin slicing machine, normal-temperature distilled water opens up piece after slice, bakes piece；
(5) slice is slightly washed with distilled water without dewaxing, directly enters haematoxylin dyeing 2min；It slightly washes, 0.5% hydrochloric acid divides rapidly
Change, 0.5% ammonium hydroxide returns indigo plant after washing；Then it is gently washed with 70% ethyl alcohol the slice several seconds, oil red O stain 10min, 70% ethyl alcohol is rapid
Break up, is dried in air, glycerin gelatine mounting.
3. the method for carrying out the dyeing of animal tissue sections grease using polyethylene glycol according to claim 1, which is characterized in that
The carbowax I is the polyethylene glycol of molecular weight 1000 ~ 2000.
4. the method for carrying out the dyeing of animal tissue sections grease using polyethylene glycol according to claim 1, which is characterized in that
The carbowax II is the polyethylene glycol mixture that molecular weight 1500 is mixed with the polyethylene glycol of molecular weight 1000 by 3:1 ~ 1:3.
5. the method for carrying out the dyeing of animal tissue sections grease using polyethylene glycol according to claim 1, which is characterized in that
The condition of step (2) and step (3) the waxdip processing are as follows: be placed in 1 ~ 3h of processing in 55 ~ 65 DEG C of incubators.
6. the method for carrying out the dyeing of animal tissue sections grease using polyethylene glycol according to claim 1, which is characterized in that
Step (3) the waxdip number of processing is 3 ~ 5 times.
7. the method for carrying out the dyeing of animal tissue sections grease using polyethylene glycol according to claim 1, which is characterized in that
The condition of step (4) the roasting piece are as follows: 2 ~ 4 h of piece is baked in 37 ~ 42 DEG C of incubators.
8. the method for carrying out the dyeing of animal tissue sections grease using polyethylene glycol according to claim 1, which is characterized in that
Animal tissue's embedded section described in step (4) with a thickness of 3 ~ 5 μm.
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| CN201910749883.9A CN110426259A (en) | 2019-08-14 | 2019-08-14 | Polyethylene glycol is used for the application of animal tissue sections grease dyeing |
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Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110361242A (en) * | 2019-08-14 | 2019-10-22 | 武汉赛维尔生物科技有限公司 | It is a kind of for the fixer of eyeball tissue and the preprocess method of eyeball tissue film-making |
| CN111929095A (en) * | 2020-07-10 | 2020-11-13 | 南京农业大学 | Small sample frozen section rapid imaging method |
| CN112611614A (en) * | 2020-11-26 | 2021-04-06 | 杭州百凌生物科技有限公司 | Preparation method and application of cell semi-solid suspension |
| CN113866199A (en) * | 2021-09-26 | 2021-12-31 | 中国海洋大学 | Method for identifying deposition part and fat cell characteristics of fish adipose tissues |
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| Publication number | Priority date | Publication date | Assignee | Title |
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