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Formaldehyde-free fixing liquid capable of being used for fixing various fresh tissues and fixing method

A technique of tissue fixation and formaldehyde fixation, applied in the field of medical histopathology, which can solve the problems of sample contamination, cumbersome operation, and heavy workload

Pending Publication Date: 2022-04-05
哈尔滨东尧科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, if one kind of fixative is used for each type of human tissue, it is necessary to prepare a large number of fixatives and mark each fixative separately, which is a huge workload.
[0004] As a kind of tissue fixative, ethanol has both fixation and dehydration effects on tissue cells. When using ethanol as a fixative, if high-concentration ethanol is directly used, the surface of the biological material will be dehydrated rapidly to form a layer similar to a protective film. structure, which affects the fixation and dehydration inside the tissue, so that the concentration of ethanol is required to have a gradient (required concentration from high to low), and the concentration gradient generally used in the prior art is 50%-70%-80%-95% - Absolute ethanol, the span of each gradient of ethanol concentration varies greatly, and the fixed dehydration effect is not good. If you want to use ethanol solutions with small spans between gradients, you need to prepare many groups of ethanol solutions with different concentrations, and frequent fixes are required. Liquid replacement, preparation, fixation, and dehydration are all cumbersome to operate, and the fixation method in the prior art is an open processing method. When the fixative needs to be replaced many times, it is easy to cause sample contamination.

Method used

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  • Formaldehyde-free fixing liquid capable of being used for fixing various fresh tissues and fixing method
  • Formaldehyde-free fixing liquid capable of being used for fixing various fresh tissues and fixing method
  • Formaldehyde-free fixing liquid capable of being used for fixing various fresh tissues and fixing method

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0075] Preparation of fixatives for various fresh tissue fixations:

[0076] Weigh raw materials:

[0077] Raw materials for preparing liquid A, by mass percentage, weigh: 15:1 picric acid saturated liquid and glacial acetic acid;

[0078] Raw materials for preparing liquid B:

[0079] The raw materials for preparing the B1 liquid are weighed in parts by mass: 10 parts of glutaraldehyde aqueous solution with a mass percentage of 25% (that is, the mass percentage of glutaraldehyde in the glutaraldehyde aqueous solution is 25%), anhydrous 40 parts of ethanol, 50 parts of phosphate buffer;

[0080] The raw materials for preparing the B2 liquid are weighed in parts by mass: 10 parts of glutaraldehyde aqueous solution with a mass percentage of 25% (that is, the mass percentage of glutaraldehyde in the glutaraldehyde aqueous solution is 25%), anhydrous 70 parts of ethanol, 20 parts of phosphate buffer;

[0081] The raw materials for preparing the B3 liquid are weighed in parts b...

Embodiment 2

[0089] The interstitial fluid that embodiment 1 is made is used for the fixation of the fresh tissue of mammary gland, comprises the following steps:

[0090] S1. Place the fresh breast tissue in liquid A, the volume ratio of liquid A to the fresh breast tissue is 20:1, perform the first fixation, and fix for 3 hours;

[0091] S2. After the first fixation is completed, wash the breast tissue with PBS buffer solution;

[0092] S3. Place the cleaned mammary gland tissue in a fixation chamber, add liquid B for second fixation, the volume ratio of liquid B to fresh mammary gland tissue is 20:1;

[0093] Specifically: first fill the fixation chamber with cleaned breast tissue with B1 solution for fixation;

[0094] After 3 hours of fixation treatment, inject new B2 solution to replace part of the used liquid, every 1 hour of fixation treatment, inject new B2 solution to replace part of the used liquid once, and replace 5 times in total;

[0095] After the fifth injection of new B...

Embodiment 3

[0098] The interstitial fluid prepared in Example 1 is used for fixing the fresh tissue of the thyroid gland, comprising the following steps:

[0099] S1. Place the fresh thyroid tissue in solution A, the volume ratio of solution A to fresh thyroid tissue is 20:1, perform the first fixation, and fix for 3 hours;

[0100] S2. After the first fixation is completed, wash the thyroid tissue with PBS buffer solution;

[0101] S3. Place the cleaned thyroid tissue in the fixation chamber, add solution B for second fixation, the volume ratio of solution B to fresh thyroid tissue is 20:1;

[0102] Specifically: first fill the fixation chamber with cleaned thyroid tissue with B1 solution for fixation;

[0103] After 3 hours of fixation treatment, inject new B2 solution to replace part of the used liquid, every 1 hour of fixation treatment, inject new B2 solution to replace part of the used liquid once, and replace 5 times in total;

[0104] After using new B2 solution for the fifth ti...

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PUM

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Abstract

The invention discloses a formaldehyde-free fixing liquid for fixing various fresh tissues, which comprises a liquid A and a liquid B, the liquid A and the liquid B are used for sequentially carrying out first fixation and second fixation on the fresh tissues, various substances in the various tissues can be effectively fixed, and the fixed tissues are not obvious in expansion and contraction, so that the tissues keep the morphological structure in a living state as far as possible; and subsequent slicing and pathological examination are facilitated.

Description

technical field [0001] The invention belongs to the technical field of medical histopathology, and in particular relates to a formaldehyde-free fixative solution and a fixation method that can be used for fixing various fresh tissues. Background technique [0002] In histopathology, it is necessary to properly and effectively fix the collected fresh tissues in time to prevent tissue autolysis and corruption, to precipitate or coagulate intracellular proteins, fats, sugars, enzymes and other components, and to make tissues and cells Keep the morphological structure in the living state as much as possible, so that the subsequent work such as slicing can be carried out smoothly. [0003] Fixative is a commonly used tissue fixative. The traditional tissue fixative is formalin (formaldehyde aqueous solution), which has the problems of volatile formaldehyde and high toxicity. It is easy to pollute the environment and cause damage to the operator's body during use. . At the same ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N1/36G01N1/28G01N1/34G01N33/00
Inventor 王金涛刘玭
Owner 哈尔滨东尧科技有限公司
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