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Histopathological tissue sample treatment method

A technology for tissue samples and processing methods, applied in sampling, scientific instruments, preparation of samples for testing, etc., can solve problems such as difficulty in pathological diagnosis, and achieve the effect of maintaining color stability time, good staining effect, and high stability

Pending Publication Date: 2020-03-17
YUNNAN UNIV OF TRADITIONAL CHINESE MEDICINE
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Because histopathological tissue samples are easily squeezed, and multi-channel staining is often required in actual work, it brings great difficulties and risks to pathological diagnosis

Method used

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Examples

Experimental program
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Effect test

Embodiment 1

[0033] A method for processing histopathological tissue samples, including the following steps:

[0034] (1) Fixation: Fix the isolated tissue sample, and fix it with Fixative I and Fixative II in sequence;

[0035] Fixing agent I is made by mixing the following components: 2.5% glutaraldehyde 50mL, 2% paraformaldehyde 50mL; Fixing agent I is fixed at 4°C, and the fixing time is 30min;

[0036] Fixing agent Ⅱ is made by mixing the following components: 40% formaldehyde 10mL, distilled water 90mL, NaH 2 PO 4 ·H 2 O 0.5g, Na 2 HPO 4 0.5g; Fixative II is fixed at room temperature for 30 minutes.

[0037] (2) Rinse: Use running water to rinse slowly.

[0038] (3) Dehydration: gradient ethanol dehydration: 75% ethanol 30 min, 80% ethanol 30 min, 85% ethanol I 30 min, 85% ethanol II 60 min, 85% ethanol III 60 min, 95% ethanol I 40 min, 95% ethanol II 40 min, anhydrous ethanol I 30 min, no Water ethanol II 30min.

[0039] (4) Transparency: select TO transparent agent for transparency, TO trans...

Embodiment 2

[0048] A method for processing histopathological tissue samples, including the following steps:

[0049] (1) Fixation: Fix the isolated tissue sample, and fix it with fixative I and fixative II in sequence;

[0050] Fixing agent I is made by mixing the following components: 2.5% glutaraldehyde 50mL, 2% paraformaldehyde 50mL; Fixing agent I is fixed at 6°C, and the fixing time is 30min;

[0051] Fixing agent Ⅱ is made by mixing the following components: 40% formaldehyde 10mL, distilled water 90mL, NaH 2 PO 4 ·H 2 O 0.5g, Na 2 HPO 4 0.5g; Fixative II is fixed at room temperature, and the fixing time is 30min.

[0052] (2) Rinse: Use running water to rinse slowly.

[0053] (3) Dehydration: gradient ethanol dehydration: 75% ethanol 30 min, 80% ethanol 30 min, 85% ethanol I 30 min, 85% ethanol II 60 min, 85% ethanol III 60 min, 95% ethanol I 40 min, 95% ethanol II 40 min, anhydrous ethanol I 30 min, no Water ethanol II 30min.

[0054] (4) Transparency: select TO transparent agent for transpa...

Embodiment 3

[0063] A method for processing histopathological tissue samples, including the following steps:

[0064] (1) Fixation: Fix the isolated tissue sample, and fix it with fixative I and fixative II in sequence;

[0065] Fixing agent I is made by mixing the following components: 2.5% glutaraldehyde 50mL, 2% paraformaldehyde 50mL; Fixing agent I is fixed at 6°C, and the fixing time is 30min;

[0066] Fixing agent Ⅱ is made by mixing the following components: 40% formaldehyde 10mL, distilled water 90mL, NaH 2 PO 4 ·H 2 O 0.5g, Na 2 HPO 4 0.5g; Fixative II is fixed at room temperature, and the fixing time is 30min.

[0067] (2) Rinse: Use running water to rinse slowly.

[0068] (3) Dehydration: gradient ethanol dehydration: 75% ethanol 30 min, 80% ethanol 30 min, 85% ethanol I 30 min, 85% ethanol II 60 min, 85% ethanol III 60 min, 95% ethanol I 40 min, 95% ethanol II 40 min, anhydrous ethanol I 30 min, no Water ethanol II 30min.

[0069] (4) Transparency: select TO transparent agent for transpa...

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Abstract

The invention provides a histopathological tissue sample treatment method, which comprises the following steps: (1) fixing: fixing an in-vitro tissue sample by adopting a fixing agent I and a fixing agent II in sequence during fixing; (2) washing; (3) dehydrating; (4) transparency treatment; (5) wax dipping and embedding; and (6) slicing and dyeing: carrying out dewaxing, treating dewaxed slices with a hematoxylin pre-dyeing solution for 2-3 minutes, and then, treating the dewaxed slices with a hematoxylin dyeing solution for 5-10 minutes, wherein the hematoxylin dyeing assistant solution comprises the following components: hematoxylin, aluminum sulfate, potassium permanganate, citric acid, nonylphenol polyoxyethylene ether and water, and the hematoxylin dyeing solution comprises the following components: hematoxylin, aluminum potassium sulfate dodecahydrate, potassium permanganate, acetic acid, nonylphenol polyoxyethylene ether, glycerol, absolute ethyl alcohol and water. The treatment method disclosed by the invention is easy to operate, high in tissue sample section integrity, good in dyeing effect, obvious in dyeing, clear in comparison and clear in cell hierarchy.

Description

Technical field [0001] The invention relates to the technical field of histopathological sample processing, in particular to a method for processing histopathological tissue samples. Background technique [0002] After a fresh sample is detached, autolysis and (or) corruption will occur, causing its structure to be destroyed, and at the same time, the cells will degenerate and autolyze microscopically. Histopathology, or histopathology, is a branch of pathology that studies the histological changes of various system organs when the human body is sick. The main research method is to slice (or smear) the isolated tissues (or cells) after processing, and then perform optical microscope observation after staining, and use the observation results to diagnose diseases. Therefore, histopathology requires not only the macroscopic aspects (such as color, volume, hardness, etc.) of the isolated sample to be as close as possible to the pre-ex vivo state, but also the microscopic aspect of ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N1/30
CPCG01N1/30G01N2001/302G01N2001/305
Inventor 石安华陈文玲朱晓松谭丽萍
Owner YUNNAN UNIV OF TRADITIONAL CHINESE MEDICINE
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