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Reticulocyte Stain
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Recticulocytes (immature erythrocytes) are slightly larger than normal red cells, and by using a supravital staining technique basic dyes such as brilliant cresyl blue, are precipitated into a meshwork (called reticulum) within the cell. This meshwork appears deep blue against a relatively unstained background. The reticulum is considered to be remnants of basophilic ribonucleoprotein (normally found in the cytoplasm) and the more mature the cell the less reticulum found.
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- Sodium citrate
- Sodium chloride
- Distilled water
- Brilliant cresyl blue
- Place 2-3 drops of the dye in glass test tube.
- Add 2-4 drops of the patient’s heparinized blood and gently mix.
- Incubate at 37°C for 15 – 20 min.
- Gently re-suspend the cells.
- Place one drop of the suspended cells on glass slide then put cover slide with gentle pressure.
- Examine the unfixed uncounterstained cells using oil immersion lens.
- Count at least 1000 red cells and calculate the per centage of reticulocytes present.
Normal RBC: Greenish blue
Reticulocytes: Greenish bluewith deep blue intracellular precipitate network.
Adults: 0.2 – 2 per cent.
Cord blood: 2 – 6 per cent.