Acid Phosphatase (ACP) Stain

Acid Phosphatase (ACP) Stain
Acid Phosphatase (ACP) Stain

Assay-kits

Acid Phosphatase (ACP) Stain

  • Product code: ACMBS-51
  • Specifications: 5Tests, 20Tests (Box)
  • Intended use: This kit is for staining bone marrow cell and blood cell smear
  • Store temperature: 2℃~8℃
  • Expiry date: 12 Months

More Details

PrincipleMain componentsSample requirementTest procedureResult interpretationMethod limitationPrecautions

Principle

Acid Phosphatese stain is based on diazotization-coupling principle. At pH 5.0, Acid Phosphatase hydrolyzes Naphthol AS-BI Phosphoric Acid and releases Naphthol AS-BI to mix with diazonium salt and form colored precipitate in cytoplasm.

Main components

Reagent compositionMain ingredient
1. Fixative (solution A)Formaldehyde
2. Fast garnet GBC base solution (solution B)GBC salt
3. Sodium nitrite solution (solution C)Sodium nitrite
4. Naphthol AS-BI phosphoric acid solution (solution D)Phosphoric naphthol AS-BI
5. Buffer solution (solution E)Phosphate
6. Methyl green solution (solution F)Methyl green
7. T artaric acid solutionL-tartaric acid

Sample requirement

  • Fresh bone marrow cell smear and blood cell smear.

Test procedure

A. Working solution preparation

Dip stain working solution preparation (50ml volume dye vat: A dip stain working solution is 43.5ml, at the same time can place at least 8-10 smears.)

  • 1. Preparation of diazonium salt solution: Each of 0.5 ml of B and C solution thoroughly mixed and let stand for 2 minutes.
  • 2. Add 40 ml of distilled water in dye vat (prepare dye vat by yourself).
  • 3. The diazonium salt solution is poured into the dye vat, and mix.
  • 4. Plus 0.5 ml of D solution and 2 ml of E solution into the dye vat, mix gently. (Add 2ml tartaric acid solution for Tartaric acid inhibition test.)
 Solution BSolution CDistilled waterSolution DSolution ETartaric acid solution
Dye vat Α0.5ml0.5ml40ml0.5ml2ml
Dye vat B (Inhibition test)0.5ml0.5ml40ml0.5ml2ml2ml

Note: The above is the preparation amount of the dip staining working solution for the 50ml volume dyeing vat. Each laboratory can scale up or down according to the volume of its own dyeing vat.

B. Drops stain working solution preparation (A drops stain working solution is 2.175ml.)

  • Apparatus required: disposable tube, micropipette, disposable tips, dropper.
  • Instruction: Mix 25μl solution B with 25μl solution C, wait for 2 minutes. Add distilled water 2ml, and then add 25μl solution D and 100μl solution E. Mix thoroughly. (Add 2 drops of tartaric acid solution for Tartaric acid inhibition test.)
 Solution BSolution CDistilled waterSolution DSolution ETartaric acid solution
Tube Α25μl25μl2ml25μl100μl
Tube B (Inhibition test)25μl25μl2ml25μl100μl2 drops

C. Staining procedure:

  • 1. Dried smears then fixed with fixative (Solution A) (Return to room temperature before use and shake well) for 30 to 60 seconds, rinse with distilled water, wait until dry.
  • 2. Add or dip in the working solution incubate at 37℃ for 60 minutes. Rinse with distilled water and dry it.
  • 3. Counterstain with solution F for 1~2 minutes. Rinse with distilled water. Air dry the smear for microscopic examination.

Result interpretation

  • Mauve granules are positive.
  • Reference range:
  • 1. Macrophages and some reticulocytes are with stronger positive reaction.
  • 2. Monocyte, T lymphocytes, and plasmacytes are with medium positive reaction.
  • 3. Granulocyte, megakaryocytes, and platelets are with positive or weakly positive reaction.
  • 4. Red blood cell system and B lymphocytes are in negative reaction.
  • 5. Acid Phosphatase are found in most tissues. Strong enzyme activity of acid phosphatase can be noted in prostatic cells.
  • 6. Hairy cell leukemia, hairy cell ACP mostly stained strongly positive or moderately positive, and not inhibited by L-tartaric acid, tartaric acid inhibited other cells were negative or very weakly positive.

Method limitation

  • Only use for morphological staining to observe.

Precautions

  • 1. 6 Tests only use on drops stain. 20 Tests and 100 Tests can be used for drip staining and dip staining.
  • 2. Return to room temperature before use and shake well (Recommend to store solution F at room temperature). Please thoroughly mixed B and solution C, all containers must be clean, the normal working solution color was dark yellow, if the working solution was light brown or dark brown, indicating that solution B and solution C is not fully mixed, which will cause dyeing failure.
  • 3. The enzyme activity of unstained smear after 24 hours of storage shall be decreasing.
  • 4. Working solution should be used within 10 minutes after preparation.
  • 5. Tightly cap the reagent bottle immediately after use to avoid evaporation.
  • 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. The PRC Ministry of Health Medical Administrative Department. National Clinical Laboratory Procedures (M) version 3. Nanjing: Southeast University Press, 2006.

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

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