Sudan Black B Stain (SBB)

Assay-kits

Product code: ACMBS-49
Specifications: 3x100mL (Box)
Intended use: This kit is for staining bone marrow cell and blood cell smear
Store temperature: 5℃～30℃
Expiry date: 12 Months

More Details

PrincipleMain componentsSample requirementTest procedureResult interpretationMethod limitationPrecautions

Principle

Sudan black B is a fat-soluble dye, lipids within the cell can be displayed, showing the Brown-black granular located in the cytoplasm.

Main components

Reagent composition	Main ingredient
1. Fixative	Formaldehyde
2. Sudan black B solution	Sudan black B
3. Wright-Giemsa solution	Wright-Giemsa
4. Phosphate buffer powder	Phosphate

Sample requirement

Fresh bone marrow cell smear and blood cell smear.

Test procedure

1. Phosphate buffer preparation: phosphate buffer powder 1 package, dissolved in 1000ml of distilled water, completely dissolved, sealed at room temperature, each time to take out.
2. The staining requires the use of dip method.
3. Dried smears of bone marrow or blood smears fixed with fixative (Return to room temperature before use and shake well) for 30 to 60 seconds, rinse with distilled water, wait until dry.
4. Smear immersed in Sudan black B solution at 37℃ for 20 to 30 minutes, rinse with distilled water, wait until dry or dry by filter paper.
5. Counterstained with Wright-Giemsa (Wright-Giemsa staining solution with phosphate buffer mixed 1:2 ratio, stained 3 to 5 minutes), rinse with distilled water, after dried then microscopy.

Result interpretation

Positive reaction was Brown-black granular, located in the cytoplasm.

Normal distribution:

1. Granulocyte system: the granulocyte system in addition to the early myeloblasts, the next phase of cells was positive reaction.
2. Monocytes system: some cells were weak positive reaction, small particles, loose, dispersively distributed, some cells were negative reaction.
3. Macrophages showed different degrees of positive reaction.
4. Lymphocytes, plasmacytes, nucleated red blood cells, megakaryocytes were negative reaction.

Method limitation

Only use for morphological staining to observe.

Precautions

1. Shake well before using the reagents, all containers must be clean.
2. Until Sudan Black B solution temperature was raised to 37℃, and then starts counting.
3. Sudan black B solution need to be dip staining, please prepare dye vat by yourself.
4. Sudan black B solution used at room temperature closed preservation, to prevent evaporation, can be used repeatedly, each time with a fresh blood can be used as a control, if the dyeing effect is weakened, the same piece can be replaced after dyeing dye again.
5. Smears may not need to be fixed, directly into the Sudan black B solution for incubation.
6. This kit must be used by professionals and the interpretation of results.
7. Should carefully read the instruction manual before using. Can only be used before expiration date, and good personal hygiene protection.
8. After use, disposal of waste should comply with the hospital or the environmental protection department requirements.
9. Production lot number and expiration date on the package.

References

1. National Health and Family Planning Commission of the People's Republic of China Medical Administration Hospital Authority. National Clinical Laboratory Procedures [M]. 4th ed. Beijing: People's Medical Publishing House, 2014.

2. Gradwohl's Clinical Laboratory Methods And Diagnosis, 8th, ed.

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