Reticulocyte Count

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Reticulocyte count:

Reticulocyte are young red cell released from the bone marrow into the blood stream and that contain RNA and ribosome but no nucleus.

After staining with a supravital dye such as new methylene blue, RNA appears as blue precipitate granules or filamentes, within the red cell.

Supravital staining refers to staining of cells in a living state before they are killed by fixating of cell or drying.

Reticulocyte count is performed by manual method.
Recently methods based on flow cytometry have been introduced which are rapid and more presice.

Principle:-
  A few drops of blood (collected in EDTA) are incubated with new methylene blue solution which stains granules of RNA in red cells.

 Number of reticulocytes is expressed as a percentage of red cell.

Specimen:-
Capillary blood or EDTA anticoagulant blood can be used.

 Procedure:-
1. Take 1-2 drops of New Methylene Blue stain / Retic stain in a test tube.
2. Add equal amount of blood mix well.
3. Keep the mixture at room temperature (37)  for 15 minutes.
4. Prepare a thin smear and allow to dry in the air.
5. Examine under microscope.
6. Count 1000 RBC and note the number of RBC that are reticulocytes.


Calculation:-

Reticulocyte percentage = number of reticulocytes counted / number of RBC counted * 100

Refference range= 0.5 % - 2.5 % in adult and children
  Reticulocyte count is higher in newborn.

Significance:- 
Diagnose Anaemia due to ineffective erythropoiesis.



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