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BACKGROUND
The reticulocytes are immature red blood cells. These are produced during erythropoiesis and maturation happens in bone marrow. They do not have cell nucleus and possess a reticular network of ribosomal RNA and are visible with methylene blue and Romanowsky stain under microscope. The number of reticulocytes indicates the healthy activity of bone marrow.1 The reticulocyte count is higher in newborn and in adults it ranges from 0.5%-2%.2
The aim of the experiment is determination of reticulocyte count.
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REQUIREMENTS
Subjects: Human
Apparatus: New methylene blue solution
Blood sample
Test tube
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PROCEDURE
Manual method
Collect the blood sample from the subject. Add methylene blue 2-3 drops to the test tube. Mix the solution with equal amount of blood and shake it well. The mixture is kept at room temperature for 15 minutes. Prepare a thin smear of the mixture and allow it to dry. Examine under the microscope.
The reticulocytes take up the methylene blue stain and immature reticulocytes show a large amount of precipitated material and the most mature reticulocytes show granular appearance. Count the number of red cells and reticulocytes.
Reticulocyte % = NR/NRBC x 100
(NR=Number of reticulocytes counted, NRBC= Number of red blood cells counted).
Absolute reticulocyte count = Reticulocyte percentage × Red cell count
Corrected reticulocyte count= Reticulocyte % × PCV of patient / Normal PCV
Reticulocyte maturation production index =corrected reticulocyte count/ Estimated maturation time in days.
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CONCLUSION
The total reticulocyte count helps in diagnosis of conditions like hemolytic anemia, chemotherapy, aplastic anemia, pernicious anemia, vitamin & mineral deficiencies,
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REFERENCES
- https://en.m.wikipedia.org/wiki/Reticulovyte. Accessed on 5 April 2018.
- https://www.bioscience.pk/component/k2/item/146-how-to-count-reticulocytes?Itemid=1857. Accessed on 5 April 2018.
