Sudan black B (SB) is a fat-soluble dye, which can be dissolved in the lipid-containing structures in the cell plasma, so that the lipids in the cytoplasm appear as brownish-black or dark black particles. This experiment is from Mudanjiang Medical College, undergraduate 5-year laboratory guide for testing majors.

Operation method

Sudan black staining experiment

Principle

Sudan black B (SB) is a fat-soluble dye that dissolves in the lipid-containing structures within the cell plasma, causing the lipids in the cytoplasm to appear as brownish-black or dark-black particles.

Materials and Instruments

Cell plasma
Formalin solution Alcoholic solution of Sudan black

Move

I. Experimental reagents:

1. formalin solution

2. Sudan Black Alcohol Solution

(1) Preparation solution: Sudan Black B 0.3 g dissolved in 100 ml of pure alcohol, at room temperature, shaking frequently, a few days after the complete dissolution of

(2) Buffer: 16 g of phenol dissolved in 30 ml of pure alcohol, and then with a mixture of 100 ml of distilled water containing 0.3 g of disodium hydrogen phosphate (Na2HPO4-12H2O).

(3) Staining solution: mix 60 ml of storage solution with 40 ml of buffer wave, filter available for several weeks.

3. 70% ethanol

4. Ricci stain

III.

l. Sufficiently dried smears were fixed in formalin vapor for 5-10 minutes;

2. Rinsed in running water for 5-10 minutes

3 . Place in Sudan black staining solution for 30 minutes.

4. Rapidly wash with 70% ethanol.

5. Rinse with tap water for 1 minute.

6. Ricci staining re-staining.

Results:

Positive reaction is brown-black or dark black granules, localized in the cytoplasm, granulocyte and monocyte series are positive. In the granulocyte series, the primary granulocytes are generally positive, or weakly positive, the cells in the stages below the early juvenile granulocytes gradually strengthen the degree of positivity as they mature, the granules of neutrophils are of uniform size and similar to the peroxidase granules; the granules of eosinophils are larger, with brown coloring and foamy sensation, and the granules of basophils are of unequal sizes. The granules in the cytoplasm of monocytes were small, irregular, and diffusely distributed, with some covering the nucleus. The lymphocyte series was negative, and the erythrocyte lineage, megakaryocytes, and platelets were negative.

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