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Method for removing melanin in tissue

A melanin and removal technology, which is applied to the field of removing melanin in tissues, can solve the problems of long decolorization time, film dropping, and easy film dropping, and achieves the effects of good removal effect, safe use and low cost.

Inactive Publication Date: 2018-02-02
青岛金域医学检验实验室有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Among them, the potassium permanganate oxalic acid method is a traditional oxidant depigmentation method, but the decolorization time of this method is long, and it is easy to fall off; the decolorization methods such as hydrogen peroxide and bromine water generally need 24-48h, and it takes a long time; the improved Although various methods can shorten the decolorization time, hydrogen peroxide or potassium permanganate is still used as the basic reagent, and the concentration or heating method is used to process, so that the reaction is too strong, and the phenomenon of falling off is not fully resolved; and, tissue After the slices are treated with the above-mentioned oxidants, the structure of the antigen may change, affecting the combination of the antigen and the antibody, which in turn will affect the antigenicity of the tissue slices; in addition, potassium permanganate is a dangerous substance in the laboratory, and it is strictly forbidden to discharge it into the environment Among them, it will explode when it comes into contact with ethanol, sulfuric acid, hydrogen peroxide, etc., and it will burn strongly when it meets glycerin.
However, there is no report on applying trichloroisocyanuric acid to remove melanin in tissues

Method used

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  • Method for removing melanin in tissue
  • Method for removing melanin in tissue

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0039] (1) Preparation of trichloroisocyanuric acid aqueous solution:

[0040] Environmental requirements: room temperature 20~35℃, humidity 30%~65%

[0041] Weigh trichloroisocyanuric acid powder or tablet (1.25g, available chlorine content reaches 500mg) with an electronic balance, dissolve in 500ml distilled water, and oscillate for 10 minutes with the ultrasonic wave sent by the ultrasonic instrument to promote the mixing of the solution and make the trichloroisocyanuric acid Cyanuric acid dissolves uniformly in water. Use a pH meter to detect the pH value of the solution, and control the pH range from 5.5 to 6.5.

[0042] (2) Preparation of tissue slices:

[0043] The pathological tissue samples to be processed were sliced ​​at 4 μm with a Leica automatic microtome, placed in a prepared 500ml trichloroisocyanuric acid solution with ordinary glass slides, removed after 20 minutes of decolorization, washed with water for 30 seconds, and routine HE staining was performed.

...

Embodiment 2

[0051] HMB45, melan A, and MITF, as specific markers of melanoma, are widely used in the auxiliary diagnosis of malignant melanoma.

[0052] In this embodiment, 0.25wt% trichloroisocyanuric acid aqueous solution, 0.25wt% potassium permanganate solution / 2% oxalic acid solution and 10% hydrogen peroxide solution were used to de-melanize the pathological samples. It was found that: after adopting 0.25wt% aqueous solution of trichloroisocyanuric acid, the signals of HMB45, melan A and MITF could be observed, and the signal intensity was not significantly different from that of the blank control; 0.25wt% potassium permanganate solution / 2 After treatment with % oxalic acid solution, the signals of melan A and MITF disappeared; after treatment with 10% hydrogen peroxide solution, the signal of melan A disappeared, and the signal intensity of HMB45 weakened.

[0053] It can be seen that the tissue antigenicity of the slices will not be affected after being treated by the demelaninizat...

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Abstract

The invention discloses a method for removing melanin in a tissue, and the method comprises the following steps: slicing a tissue specimen to be processed, then placing the tissue specimen in a solution containing trichloroisocyanuric acid as an effective component, decoloring for 20-30min, washing with water to complete removal of the melanin in the tissue. The method finds for the first time that during the preparation of pathological tissue sections, the melanin deposited in the tissue specimen can be quickly removed by use of a low concentration of solution containing the trichloroisocyanuric acid, removal conditions are mild, and the removal of the melanin can be carried out at normal temperature without heating of increase of remover concentration.

Description

technical field [0001] The invention relates to the technical field of preparation of pathological tissue slices, in particular to a method for removing melanin in tissues. Background technique [0002] Specimens containing melanin deposits are often encountered in pathological routine tissue work. Melanin is common in malignant melanoma, pigmented nevus, skin disease, pigmented neurofibroma, clear cell sarcoma, etc., especially in malignant melanoma, a large amount of melanin often affects the observation of cell nucleus and morphology, especially the application of DAB in immunohistochemistry Therefore, it is an important measure to depigment the melanin deposited in the tissue. [0003] Melanin is a self-derived pigment produced by human and animal melanocytes. Tyrosine forms dopa under the action of tyrosinase and then produces melanin. It is characterized by relatively stable physical and chemical properties, and is not easy to dissolve in water and organic solutions. ...

Claims

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Application Information

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IPC IPC(8): G01N1/28G01N1/30
Inventor 张龙翔辛泽明孙显路金卫中侯生根刘超广薛彦青
Owner 青岛金域医学检验实验室有限公司
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