Leukocyte Peroxidase (POX) (Oxidation WG-KI) Stain

Assay-kits

Leukocyte Peroxidase (POX) (Oxidation WG-KI) Stain

Product code: ACMBS-42
Specifications: 5Tests, 20Tests, 100Tests (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

Peroxidases in cells dissolve oxides to generate oxygen. Oxygen shall react with to produce iodine. Iodine works with Wright Giemsa stain and forms colored granules in cytoplasm.

Main components

Reagent composition	Main ingredient
1. Solution A	Eosin
2. Solution B	Azure II
3. KI Solution (solution C)	Potassium iodide
4. WG Solution (solution D)	Wright-Giemsa

Sample requirement

Fresh bone marrow cell smear and blood cell smear.

Test procedure

This method uses drop staining, drop stain working solution preparation (One part of drop stain working solution is 2.4ml).

Apparatus required: disposable tube, micropipette, disposable tips, dropper.
Instruction: Mix well the tube while adding 2.0ml solution C to 400μl solution D. Use the working solution within 2 hours.
Procedure:

1. Rapid stain (nuclear staining): Add solution B (solution A: solution B = 1:1 or 1:2) immediately after solution A is added to dried bone marrow smear. Stain for 20～30 seconds, rinse with tap water. Remove the excess water or dry with filter paper.
2. Drop working solution on the slide and stain for 40 seconds to 1 minute. Throw off the stain (Do not rinse with water.), dry with filter paper and examine under a microscope.

Result interpretation

In positive reaction, red-brown to dark blue granules shall be found in cytoplasm. Red-brown granules are with weakly positive reaction. Dark purple to dark blue granules spread over cytoplasm and even fully cover nuclei in strong positive reaction. Blue cytoplasm means negative, and no positive granules are found, while the nuclei shall be evenly stained as red-purple. Eosinophils can be quickly stained in vivid color, and its positive reaction is dark blue. Some cellular granules shall spread diffusely to the outside of cell. Acicula shall appear in the outer layer of cells.

Normal Distribution

1. Granulocyte System: Neutrophils other than myelobalsts and cells at lower stages are positive. Eosinophils are found to be stained fast and deeply, basophil are in negative reaction.
2. Monocytes System: Some cells are with weakly positive reaction and have small granules diffusely spread in cells. Some cells are in negative reaction.
3. Macrophages are in positive reaction at variant degrees.
4. Lymphocytes, plasmacytes, red blood cells, and megakaryocytes are negative.

Method limitation

Only use for morphological staining to observe.

Precautions

1. Return to room temperature before use, shake well before using reagents, used containers must be clean.
2. If a large number of cells appear, POX reaction shall become weak or lead to bad staining results. Restain the smear with working solution is thus suggested to improve the performance.
3. Do not rinse with water, since positive granules resulted from potassium iodide POX Stain are easily dissolved by water.
4. Tightly cap the reagent bottle immediately after use to avoid evaporation.
5. This kit must be used by professionals and the interpretation of results.
6. Should carefully read the instruction manual before using. Can only be used before expiration date, and good personal hygiene protection.
7. After use, disposal of waste should comply with the hospital or the environmental protection department requirements.
8. 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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