Retic stain

Histopathology
 Reticulocyte count
VALUE OF TEST

 Reticulocytes are immature red cells normally present in small numbers in the
blood (up to 2%).
 Reticulocyte numbers increase when there is an increase in erythropoietic activity.
 A reticulocyte count assesses bone marrow activity, e.g. whether there is an
effective erythropoietic response when there is a reduction in the number of red
cells due to haemolysis or haemorrhage.
 A reticulocyte count is also of value in monitoring the erythropoietic response of an
anaemic patient following treatment
 PRINCIPLE OF TEST
 A supravital stain (i.e. one that stains living material) such as New
methylene blue or brilliant cresyl blue is incubated with a few drops of
blood.
 To detect ribosomal RNA in reticulocytes, the red cells must be stained
while they are still living (not fixed).
 A thin preparation is made and the reticulocytes counted microscopically.
 Reticulocytes are recognized by the violet-blue stained granules of
ribosomal RNA (reticulin).
 The reticulocyte count is usually expressed as a percentage
REAGENT
New methylene Reagent or blue or brilliant cresyl blue

SPECIMEN
Use well-mixed EDTA anticoagulated blood or if the patient is a young
child, use capillary blood
Procedure

By self
COUNTING RETICULOCYTE
1. Count a total of 500 red cells, noting on paper the number of cells
that are reticulocytes.
2. Multiply the number of reticulocytes counted by 2.
3. Divide the figure by 10 to obtain the percentage figure.
INTERPRETATION OF RETICULOCYTE COUNT
1. Infants at birth = 2–5%
2. Children and adults = 0.5–2.5%
RAISED RETICULOCYTE COUNTS:
Found when there is an increase in red blood cell production as occurs
in:
1. Haemolytic anaemias (with effective erythropoiesis).
2. Following acute blood loss
3. After iron therapy for iron deficiency anaemia or specific therapy for
megaloblastic anaemia.
DECREASED RETICULOCYTE COUNT:
Associated with ineffective erythropoiesis or decreased production of
red cells.