Copper Stain (Rubeanic Acid Method)

Assay-kits

Copper Stain (Rubeanic Acid Method)

Product code: ACMBS-41
Specifications: 5Tests, 20Tests (Box)
Intended use: Used to stain the copper particles in the tissue
Store temperature: 2℃～8℃
Expiry date: 18 Months

More Details

PrincipleMain componentsSample requirementTest procedureResult interpretationPrecautions

Principle

Hepatolenticular degeneration, also known as Wilson disease, is an autosomal recessive copper metabolic disorder. Due to the poor specificity of liver lesions in Wilson's disease histologically, copper staining in liver tissue is very important for the differentiation and diagnosis of Wilson's disease. Copper and some other metals can form chelates with erythrocytic acid. Adding sodium acetate can blocking-up the binding of red acid and other metals other than copper.

Main components

Reagent composition	Main ingredient
1. Rubeanic acid solution (solution A)	Rubeanic acid
2. Sodium acetate solution (solution B)	Anhydrous sodium acetate
3. Nuclear fast red solution	Nuclear fast red

Sample requirement

The tissue section must be fully fixed.

Test procedure

A. Working solution preparation:

Mix solution A and solution B at 1:20 (Mix before use).

B. Staining procedure:

1. The slices are routinely dewaxed to water and washed once with distilled water.
2. The sections were immersed in the working solution and stained at 37℃ incubator for.
3. 18～24 hours or in a 60℃water bath for 1～2 hours.
4. Take out slice, 75% ethanol was added into the slices and treated at room temperature for 30 minutes.
5. The slices were immersed in anhydrous ethanol for 10 minutes, distilled water washing.
6. Nuclear solid red dye staining for 1～2 minutes, and washing.
7. Conventional dehydration and transparent, sealed with neutral resin.

Result interpretation

Copper particles	Dark green
Nucleuses	Red

Precautions

1. When the tissue sections were put into the working fluid at 37℃ incubator overnight, the amount of staining solution should be sufficient, the staining temperature would be constant, and the staining should be in plenty of time.
2. Due to the long staining time, care should be taken to prevent the tissue from detaching during the staining process, especially if the water bath is at 60℃. Pay attention to the cleanness of the slides. It is best to use slide-proof slides. The spreading slides must be flat and free of wrinkles. The temperature and time of baking slides must be sufficient.
3. The working solution can be reused 2～3 times. After each use, it must be sealed and stored at 2～8℃ in time to prevent the working solution from evaporating or deteriorating. The validity period is 1 month.
4. This reagent should be used by professionals and interpretation of the results. Read the instruction manual carefully before use, use it within the validity period, and do personal hygiene protection.
5. Dispose of waste according to the requirements of the hospital or environmental protection department after use.
6. The production date, production batch number and expiration date are shown in the outer packaging.

References

1. Sun Yanling, Zhao Jingmin, Yang Jianfa, et al. Clinical characteristics of 48 children with Wilson's disease and the mechanism of liver fibrosis [J]. Chinese Journal of Medicine. 2005.30(4): 300-32.

2. Zhao Yulai, Yang Jianfa, Pan Deng, and so on. The application of copper staining in the pathological diagnosis of liver disease in Wilson's disease. The 302 Hospital of the People's Liberation Army.

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