CN110973116A - Adipose tissue fixing liquid and preparation method and application method thereof - Google Patents
Adipose tissue fixing liquid and preparation method and application method thereof Download PDFInfo
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- CN110973116A CN110973116A CN201911335026.0A CN201911335026A CN110973116A CN 110973116 A CN110973116 A CN 110973116A CN 201911335026 A CN201911335026 A CN 201911335026A CN 110973116 A CN110973116 A CN 110973116A
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- 210000000577 adipose tissue Anatomy 0.000 title claims abstract description 78
- 238000000034 method Methods 0.000 title claims abstract description 26
- 238000002360 preparation method Methods 0.000 title claims abstract description 13
- 239000007788 liquid Substances 0.000 title abstract description 16
- 210000001519 tissue Anatomy 0.000 claims abstract description 34
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims abstract description 30
- WSFSSNUMVMOOMR-UHFFFAOYSA-N Formaldehyde Chemical compound O=C WSFSSNUMVMOOMR-UHFFFAOYSA-N 0.000 claims abstract description 27
- QTBSBXVTEAMEQO-UHFFFAOYSA-N Acetic acid Chemical compound CC(O)=O QTBSBXVTEAMEQO-UHFFFAOYSA-N 0.000 claims abstract description 18
- YGSDEFSMJLZEOE-UHFFFAOYSA-N salicylic acid Chemical compound OC(=O)C1=CC=CC=C1O YGSDEFSMJLZEOE-UHFFFAOYSA-N 0.000 claims abstract description 18
- 238000007598 dipping method Methods 0.000 claims abstract description 16
- 229920001223 polyethylene glycol Polymers 0.000 claims abstract description 11
- UXVMQQNJUSDDNG-UHFFFAOYSA-L Calcium chloride Chemical compound [Cl-].[Cl-].[Ca+2] UXVMQQNJUSDDNG-UHFFFAOYSA-L 0.000 claims abstract description 9
- 239000002202 Polyethylene glycol Substances 0.000 claims abstract description 9
- 229960000583 acetic acid Drugs 0.000 claims abstract description 9
- BFNBIHQBYMNNAN-UHFFFAOYSA-N ammonium sulfate Chemical compound N.N.OS(O)(=O)=O BFNBIHQBYMNNAN-UHFFFAOYSA-N 0.000 claims abstract description 9
- 229910052921 ammonium sulfate Inorganic materials 0.000 claims abstract description 9
- 235000011130 ammonium sulphate Nutrition 0.000 claims abstract description 9
- 229910001628 calcium chloride Inorganic materials 0.000 claims abstract description 9
- 239000001110 calcium chloride Substances 0.000 claims abstract description 9
- 239000012362 glacial acetic acid Substances 0.000 claims abstract description 9
- FJKROLUGYXJWQN-UHFFFAOYSA-N papa-hydroxy-benzoic acid Natural products OC(=O)C1=CC=C(O)C=C1 FJKROLUGYXJWQN-UHFFFAOYSA-N 0.000 claims abstract description 9
- 229960004889 salicylic acid Drugs 0.000 claims abstract description 9
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims abstract description 9
- 208000005156 Dehydration Diseases 0.000 claims abstract description 7
- 230000018044 dehydration Effects 0.000 claims abstract description 7
- 238000006297 dehydration reaction Methods 0.000 claims abstract description 7
- 239000012188 paraffin wax Substances 0.000 claims description 14
- CTQNGGLPUBDAKN-UHFFFAOYSA-N O-Xylene Chemical compound CC1=CC=CC=C1C CTQNGGLPUBDAKN-UHFFFAOYSA-N 0.000 claims description 11
- 239000001993 wax Substances 0.000 claims description 10
- 238000002844 melting Methods 0.000 claims description 6
- 230000008018 melting Effects 0.000 claims description 6
- 239000008118 PEG 6000 Substances 0.000 claims description 5
- 229920002584 Polyethylene Glycol 6000 Polymers 0.000 claims description 5
- MTHSVFCYNBDYFN-UHFFFAOYSA-N diethylene glycol Chemical compound OCCOCCO MTHSVFCYNBDYFN-UHFFFAOYSA-N 0.000 claims description 5
- 239000000834 fixative Substances 0.000 claims description 5
- 229920001030 Polyethylene Glycol 4000 Polymers 0.000 claims description 3
- 239000003795 chemical substances by application Substances 0.000 claims description 2
- 239000012024 dehydrating agents Substances 0.000 claims description 2
- WSFSSNUMVMOOMR-NJFSPNSNSA-N methanone Chemical compound O=[14CH2] WSFSSNUMVMOOMR-NJFSPNSNSA-N 0.000 claims description 2
- 239000012530 fluid Substances 0.000 claims 2
- 238000003778 tissue fixation method Methods 0.000 claims 1
- 239000008096 xylene Substances 0.000 claims 1
- 231100000053 low toxicity Toxicity 0.000 abstract 1
- 206010024612 Lipoma Diseases 0.000 description 10
- 230000000694 effects Effects 0.000 description 8
- 230000000052 comparative effect Effects 0.000 description 7
- 210000004027 cell Anatomy 0.000 description 5
- 102000004169 proteins and genes Human genes 0.000 description 4
- 108090000623 proteins and genes Proteins 0.000 description 4
- 241001391944 Commicarpus scandens Species 0.000 description 2
- 210000001789 adipocyte Anatomy 0.000 description 2
- 230000008602 contraction Effects 0.000 description 2
- 230000007547 defect Effects 0.000 description 2
- 238000003745 diagnosis Methods 0.000 description 2
- 238000010827 pathological analysis Methods 0.000 description 2
- 230000001988 toxicity Effects 0.000 description 2
- 231100000419 toxicity Toxicity 0.000 description 2
- 230000037303 wrinkles Effects 0.000 description 2
- 230000009286 beneficial effect Effects 0.000 description 1
- 230000007717 exclusion Effects 0.000 description 1
- 231100001261 hazardous Toxicity 0.000 description 1
- 230000036571 hydration Effects 0.000 description 1
- 238000006703 hydration reaction Methods 0.000 description 1
- 150000002632 lipids Chemical class 0.000 description 1
- 239000000203 mixture Substances 0.000 description 1
- 230000000149 penetrating effect Effects 0.000 description 1
- 230000035515 penetration Effects 0.000 description 1
- 229920000642 polymer Polymers 0.000 description 1
- 238000004804 winding Methods 0.000 description 1
Classifications
-
- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01N—PRESERVATION OF BODIES OF HUMANS OR ANIMALS OR PLANTS OR PARTS THEREOF; BIOCIDES, e.g. AS DISINFECTANTS, AS PESTICIDES OR AS HERBICIDES; PEST REPELLANTS OR ATTRACTANTS; PLANT GROWTH REGULATORS
- A01N1/00—Preservation of bodies of humans or animals, or parts thereof
-
- 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
Abstract
The invention discloses adipose tissue fixing liquid and a preparation method and an application method thereof. The preparation method of the adipose tissue fixing liquid comprises the following steps: and (2) uniformly mixing 32-36 parts of ethanol and 34-36 parts of salicylic acid, adding 18-22 parts of formaldehyde and 3-5 parts of glacial acetic acid, and continuously uniformly mixing to obtain a solution a. And (3) ultrasonically mixing 10-14 parts of polyethylene glycol, 2-5 parts of calcium chloride, 6-10 parts of ammonium sulfate and 300-400 parts of water uniformly, and storing at 4 ℃ overnight to obtain a solution b. And mixing the prepared solution a and the solution b by using a magnetic stirrer, and uniformly mixing to obtain the adipose tissue fixing solution. The fixing method comprises the following steps: and (3) taking a fat tissue sample, adding the fat tissue fixing solution, fixing for 3-24 hours at room temperature, and then performing dehydration treatment, transparent treatment and wax dipping treatment by using a full-automatic dehydrator. The adipose tissue fixing solution has the advantages of simple preparation method, low toxicity, easy operation and better application prospect.
Description
Technical Field
The invention relates to the field of biotechnology, in particular to adipose tissue fixing liquid and a preparation method and an application method thereof.
Background
In the pathological diagnosis process, specimen preparation is often required. The fixation of the specimen is crucial to the correctness of pathological diagnosis. For the fixation of adipose tissues, due to the special structure, various factors often appear in the flaking process, so that the flaking effect of specimens is poor, and the subsequent observation and diagnosis are influenced.
Adipose tissue contains a large amount of lipid components and cannot be completely fixed and dehydrated. And when fixing the adipose tissues, the fixing liquid needs to be considered to be incapable of causing hardening and shrinking of the adipose tissues, otherwise, the tabletting effect is influenced.
In the prior art, adipose tissues are generally fixed by using adipose tissue fixing liquid taking formaldehyde as a main component, and the using effect of the fixing liquid can meet the requirements of most flaking. However, formaldehyde is highly hazardous and is prone to side effects that can lead to poor fixation.
Therefore, there is a need for further improvement of the existing adipose tissue fixative, so that it can achieve better fixation effect and reduce its harmfulness.
Disclosure of Invention
In order to overcome the defects in the prior art, the embodiment of the invention provides the adipose tissue fixing liquid, and the preparation method and the application method thereof, wherein the preparation method is simple, low in toxicity, easy to operate, high in penetration speed, uniform in fixation, clear in display of the microstructure of cells, and good in application prospect.
The invention provides adipose tissue fixing liquid which comprises the following components in parts by weight:
10-14 parts of polyethylene glycol,
18-22 parts of formaldehyde,
2-5 parts of calcium chloride,
32-36 parts of ethanol,
3-5 parts of glacial acetic acid,
34-36 parts of salicylic acid,
6-10 parts of ammonium sulfate,
300-400 parts of water.
Preferably, the polyethylene glycol comprises one or more of PEG 4000, PEG6000 and PEG 10000.
The invention also provides a preparation method of the adipose tissue fixing liquid, which comprises the following steps:
uniformly mixing 32-36 parts of ethanol and 34-36 parts of salicylic acid, adding 18-22 parts of formaldehyde and 3-5 parts of glacial acetic acid, and continuously uniformly mixing to obtain a solution a;
ultrasonically mixing 10-14 parts of polyethylene glycol, 2-5 parts of calcium chloride, 6-10 parts of ammonium sulfate and 300-400 parts of water uniformly, and storing at 4 ℃ overnight to obtain a solution b;
and mixing the solution a and the solution b by using a magnetic stirrer, and uniformly mixing to obtain the adipose tissue fixing solution.
Preferably, the ultrasonic mixing frequency in the second step is 40kHz, the power is 330W-410W, the mixing time is 20-30 min, and the mixing temperature is 40-60 ℃.
Preferably, the operating speed of the magnetic stirrer in the third step is 150 rpm-250 rpm, and the temperature is room temperature.
The invention also provides a fixing method of the adipose tissues, which comprises the following steps:
taking a fat tissue sample, adding a fat tissue fixing solution, and fixing for 3-24 hours at room temperature;
and (3) carrying out dehydration treatment, transparent treatment and wax dipping treatment on the fixed adipose tissue sample by using a full-automatic dehydrator.
Preferably, the dehydrating agent for dehydrating in the fixing method is 70-90 wt% of ethanol, and the time is 1-1.5 h.
Preferably, the clearing agent for clearing treatment in the fixing method is dimethylbenzene, and the treatment time is 30-40 min.
Preferably, in the fixing method, the wax dipping treatment adopts melted paraffin wax to dip for 3 times, each time lasts for 1-2 hours, the wax dipping temperature is 60 ℃, and the melting point of the paraffin wax is 56 ℃.
Preferably, in the fixing method, after the adipose tissue sample is taken, the volume of the added adipose tissue fixing liquid is 6-10 times of the adipose tissue volume.
The invention has the following beneficial effects:
the adipose tissue fixative is simple in preparation method, low in toxicity, easy to operate, high in penetrating speed, uniform in fixation, clear in display of the microstructure of cells and good in application prospect.
After the adipose tissue fixing liquid is adopted and the lipoma tissue is processed by the fixing method provided by the invention, when the lipoma tissue is sliced, the sample has moderate hardness and good cutting performance, the sliced tissue is smooth and not easy to break, the sliced tissue can be smoothly sliced, and the sliced tissue is easy to flatten without wrinkles, has complete tissue structure under a mirror, has clear adipose cell frame and does not have the phenomena of cell contraction, collapse and overlapping.
In order to make the aforementioned and other objects, features and advantages of the invention more comprehensible, preferred embodiments accompanied with figures are described in detail below.
Detailed Description
The technical solutions in the embodiments of the present invention will be clearly and completely described below with reference to the embodiments of the present invention, and it is obvious that the described embodiments are only a part of the embodiments of the present invention, and not all of the embodiments. All other embodiments, which can be derived by a person skilled in the art from the embodiments given herein without making any creative effort, shall fall within the protection scope of the present invention.
The adipose tissue samples used in examples 1 to 3 and comparative example 1 of the present invention were lipoma tissues.
The fixing time of the fat tissue fixing liquid to the fat tissue is determined according to different types, properties and sizes of the tissue. Since the samples used in examples 1 to 3 of the present invention and comparative example 1 were lipoma tissues, the fixation time was 10 hours.
Example 1
Preparing an adipose tissue fixing solution:
uniformly mixing 36 parts of ethanol and 34 parts of salicylic acid, adding 22 parts of formaldehyde and 3 parts of glacial acetic acid, and continuously and uniformly mixing to obtain a solution a;
ultrasonically mixing 14 parts of PEG 4000, 2 parts of calcium chloride, 10 parts of ammonium sulfate and 300 parts of water uniformly, wherein the ultrasonic mixing frequency is 40kHz, the power is 410W, the mixing time is 20min, the mixing temperature is 60 ℃, and then storing overnight at the temperature of 4 ℃ to obtain a solution b;
and mixing the prepared solution a and solution b by using a magnetic stirrer, wherein the working speed of the magnetic stirrer is 150rpmrpm, the temperature is room temperature, and uniformly mixing to obtain the adipose tissue fixing solution.
Fixation of adipose tissue:
after an adipose tissue sample is taken, adding an adipose tissue fixing solution, wherein the volume of the added adipose tissue fixing solution is 10 times of that of the adipose tissue, and then fixing for 10 hours at room temperature;
and (2) placing the fixed adipose tissue sample in a full-automatic dehydrator for treatment, dehydrating by using 70 wt% of ethanol for 1.5h, then performing transparent treatment by using dimethylbenzene for 30min, and then performing wax dipping treatment by using melted paraffin for 3 times, wherein the wax dipping temperature is 60 ℃ and the melting point of the paraffin is 56 ℃ for 2h each time.
Example 2
Preparing an adipose tissue fixing solution:
uniformly mixing 32 parts of ethanol and 36 parts of salicylic acid, adding 18 parts of formaldehyde and 5 parts of glacial acetic acid, and continuously and uniformly mixing to obtain a solution a;
ultrasonically mixing 10 parts of PEG10000, 5 parts of calcium chloride, 6 parts of ammonium sulfate and 400 parts of water uniformly, wherein the ultrasonic mixing frequency is 40kHz, the power is 330W, the mixing time is 30min, the mixing temperature is 40 ℃, and then storing overnight at the temperature of 4 ℃ to obtain a solution b;
and mixing the prepared solution a and solution b by using a magnetic stirrer, wherein the working speed of the magnetic stirrer is 250rpm, the temperature is room temperature, and uniformly mixing to obtain the adipose tissue fixing solution.
Fixation of adipose tissue:
after the adipose tissue sample is taken, adding adipose tissue fixing liquid, wherein the volume of the added adipose tissue fixing liquid is 6 times of that of the adipose tissue, and then fixing for 10 hours at room temperature;
and (2) placing the fixed adipose tissue sample in a full-automatic dehydrator for treatment, performing dehydration treatment by adopting 90 wt% of ethanol for 1h, performing transparent treatment by adopting dimethylbenzene for 40min, performing wax dipping treatment by adopting melted paraffin for 3 times, wherein the wax dipping temperature is 60 ℃ and the melting point of the paraffin is 56 ℃ for 1h each time.
Example 3
Preparing an adipose tissue fixing solution:
uniformly mixing 34 parts of ethanol and 35 parts of salicylic acid, adding 15 parts of formaldehyde and 4 parts of glacial acetic acid, and continuously and uniformly mixing to obtain a solution a;
ultrasonically mixing 12 parts of PEG6000, 3.5 parts of calcium chloride, 8 parts of ammonium sulfate and 350 parts of water uniformly, wherein the ultrasonic mixing frequency is 40kHz, the power is 370W, the mixing time is 25min, the mixing temperature is 50 ℃, and then storing overnight at the temperature of 4 ℃ to obtain a solution b;
and mixing the prepared solution a and solution b by using a magnetic stirrer, wherein the working speed of the magnetic stirrer is 200rpm, the temperature is room temperature, and uniformly mixing to obtain the adipose tissue fixing solution.
Fixation of adipose tissue:
after an adipose tissue sample is taken, adding an adipose tissue fixing solution, wherein the volume of the added adipose tissue fixing solution is 8 times of that of the adipose tissue, and then fixing for 10 hours at room temperature;
and (2) placing the fixed adipose tissue sample in a full-automatic dehydrator for treatment, performing dehydration treatment by adopting 80 wt% of ethanol for 1h, performing transparent treatment by adopting dimethylbenzene for 35min, performing wax dipping treatment by adopting melted paraffin for 3 times, wherein the dipping temperature is 60 ℃ and the melting point of the paraffin is 56 ℃ for 1.5h each time.
Comparative example 1
This comparative example differs from example 1 in that the PEG6000 component is not included. The method comprises the following specific steps:
preparing an adipose tissue fixing solution:
uniformly mixing 34 parts of ethanol and 35 parts of salicylic acid, adding 15 parts of formaldehyde and 4 parts of glacial acetic acid, and continuously and uniformly mixing to obtain a solution a;
3.5 parts of calcium chloride, 8 parts of ammonium sulfate and 350 parts of water are taken and ultrasonically mixed uniformly, the ultrasonic mixing frequency is 40kHz, the power is 370W, the mixing time is 25min, the mixing temperature is 50 ℃, and then the mixture is placed in an environment at 4 ℃ for storage overnight to obtain a solution b;
and mixing the prepared solution a and solution b by using a magnetic stirrer, wherein the working speed of the magnetic stirrer is 200rpm, the temperature is room temperature, and uniformly mixing to obtain the adipose tissue fixing solution.
Fixation of adipose tissue:
after an adipose tissue sample is taken, adding an adipose tissue fixing solution, wherein the volume of the added adipose tissue fixing solution is 8 times of that of the adipose tissue, and then fixing for 10 hours at room temperature;
and (2) placing the fixed adipose tissue sample in a full-automatic dehydrator for treatment, performing dehydration treatment by adopting 80 wt% of ethanol for 1h, performing transparent treatment by adopting dimethylbenzene for 35min, performing wax dipping treatment by adopting melted paraffin for 3 times, wherein the dipping temperature is 60 ℃ and the melting point of the paraffin is 56 ℃ for 1.5h each time.
The adipose tissue samples prepared in examples 1 to 3 and comparative example 1 were sliced and stained, and the treatment effect was observed, with the results as shown in table 1 below:
TABLE 1
| Hardness of | Quality of slice | Under mirror effect | |
| Example 1 | Moderate hardness | Good cutting performance, smooth slice and difficult breakage | Under-mirror partThe structure of fat cells is less distinct |
| Example 2 | Is slightly soft | Poor cutting performance, smooth and smooth slices and difficult breakage | Partial incompleteness of the tissue structure under the mirror |
| Example 3 | Moderate hardness | Good cutting performance, smooth slice and difficult breakage | The tissue structure under the mirror is complete and clear |
| Comparative example 1 | Is soft | Unsmooth slicing and difficult slicing | Subscopic tissue deformity and fold |
As can be seen from Table 1, after the adipose tissue fixing solution and the adipose tissue treating method are adopted to treat the adipose tissue, the adipose tissue can be well fixed, so that the treated adipose tissue can be conveniently sliced and subsequently observed, and better diagnosis can be carried out.
The adipose tissue fixative, preparation method and use method in example 3 were optimized. After the technical scheme in embodiment 3 is adopted to treat the lipoma tissue, when the lipoma tissue is sliced, the hardness of the sample is moderate, the lipoma tissue has good cutting performance, the slice is flat and is not easy to break, the lipoma tissue can be smoothly sliced, and the lipoma tissue is easy to flatten without wrinkles after being sliced, has a complete tissue structure under a mirror, has a clear lipoma frame, and does not have the phenomena of cell contraction, collapse and overlapping.
The principle of adipose tissue fixative is to precipitate proteins, causing the tissue to contract, but not harden the tissue to enable easy sectioning. After sectioning, it is also required that the fine structure of the cells be clearly shown. In this example, a polyethylene glycol component was added. Polyethylene glycol is a nonionic linear macromolecular polymer with strong hydrophilicity, so that a hydration layer can be damaged to dehydrate protein molecules, and the protein molecules are extruded by space exclusion and are precipitated by winding the protein molecules according to the chain length.
The fat tissue fixing solution of comparative example 1 did not contain PEG6000, and its slicing effect was poor. When slicing, the tissues feel soft, the slicing is not smooth, the phenomena of tissue defect and folding under the lens are serious, and the slicing quality is poor.
The principle and the implementation mode of the invention are explained by applying specific embodiments in the invention, and the description of the embodiments is only used for helping to understand the method and the core idea of the invention; meanwhile, for a person skilled in the art, according to the idea of the present invention, there may be variations in the specific embodiments and the application scope, and in summary, the content of the present specification should not be construed as a limitation to the present invention.
Claims (10)
1. The fat tissue fixing solution is characterized by comprising the following components in parts by weight:
10-14 parts of polyethylene glycol,
18-22 parts of formaldehyde,
2-5 parts of calcium chloride,
32-36 parts of ethanol,
3-5 parts of glacial acetic acid,
34-36 parts of salicylic acid,
6-10 parts of ammonium sulfate,
300-400 parts of water.
2. The fat tissue fixation fluid of claim 1, wherein the polyethylene glycol comprises one or more of PEG 4000, PEG6000, and PEG 10000.
3. A preparation method of an adipose tissue fixing fluid is characterized by comprising the following steps:
uniformly mixing 32-36 parts of ethanol and 34-36 parts of salicylic acid, adding 18-22 parts of formaldehyde and 3-5 parts of glacial acetic acid, and continuously uniformly mixing to obtain a solution a;
ultrasonically mixing 10-14 parts of polyethylene glycol, 2-5 parts of calcium chloride, 6-10 parts of ammonium sulfate and 300-400 parts of water uniformly, and storing at 4 ℃ overnight to obtain a solution b;
and mixing the solution a and the solution b by using a magnetic stirrer, and uniformly mixing to obtain the adipose tissue fixing solution.
4. The method for preparing the fat tissue fixing solution according to claim 3, wherein the frequency of ultrasonic mixing in the second step is 40kHz, the power is 330W-410W, the mixing time is 20-30 min, and the mixing temperature is 40-60 ℃.
5. The method for preparing an adipose tissue-derived fixative according to claim 3, wherein the magnetic stirrer operates at 150rpm to 250rpm in the third step and the temperature is room temperature.
6. A method for fixing adipose tissues, which comprises using the adipose tissue fixing solution according to claim 1, wherein the method comprises:
taking a fat tissue sample, adding a fat tissue fixing solution, and fixing for 3-24 hours at room temperature;
and (3) carrying out dehydration treatment, transparent treatment and wax dipping treatment on the fixed adipose tissue sample by using a full-automatic dehydrator.
7. The method for fixing adipose tissues according to claim 6, wherein the dehydrating agent used in the dehydration treatment in the fixation method is 70 to 90 wt% ethanol for 1 to 1.5 hours.
8. The method for fixing adipose tissues according to claim 6, wherein the clearing agent for the clearing treatment is xylene, and the treatment time is 30 to 40min.
9. The adipose tissue fixation method according to claim 6, wherein the paraffin dipping treatment is performed by dipping melted paraffin for 3 times, each time for 1-2 hours, the paraffin dipping temperature is 60 ℃, and the melting point of the paraffin is 56 ℃.
10. The method according to claim 6, wherein the volume of the adipose tissue-derived fixing solution added after the adipose tissue sample is extracted is 6 to 10 times the volume of the adipose tissue.
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