Topic 2: Reticulocyte Count

Learning Objectives

At the end of this topic, students will be able to:

 Describe the normal distribution and replacement pattern of reticulocytes in the

circulation.
 Compare the morphological appearances of reticulocytes stained with Wright stain and
a supravital stain, such as new methylene blue.
 Discuss various formulas used in counting reticulocytes.
 Calculate for reticulocyte counts using the different formulas
 Explain the methods of reticulocyte counting

Activating Prior Learning

Answer/do the following:

1. What is the clinical use of reticulocyte count?
2. ________ is the anticoagulant of choice for reticulocyte count.
3. ________ is the general indicator of the rate of effective erythropoiesis in patients
w/ anemia.
4. ________ used to correct the reticulocyte counts in patients with
anemia
5. Miller disc is composed of two squares; _________ are counted in the smaller
square (B), and _________ are counted in the larger square (A).

Presentation of Contents

Basic Information
} Assess erythropoeitic activity of the Bone
marrow
} Anticoagulant of choice: EDTA
} Normal Reticulocyte Count:
Adult: 0.5 to 1.5%
Newborn: 2 to 6%
} Principle: Whole blood is stained with
supravital stain. Any non-nucleated red

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 cell that contains 2 or more particles of blue stained granufilamentous material is
a reticulocyte.

Formula
1. For Routine Microscopy
¿ of retics counted
Reticulocyte ( % )= x 100
1000(RBC )

2. Using Miller Disc

} inserted into the eyepiece of the
microscope and the grid is seen
} composed of two squares; RBCs are
counted in the smaller square (B), and
reticulocytes are counted in the larger
square (A).
} Formula:

Total Retics∈Square A
Reticulocyte ( % )= x 100
Total RBC ∈Square B x 9

3. Absolute Reticulocyte Count

} is the actual number of reticulocytes in 1 liter (L) or 1 microliter (mL) of
blood
} Formula:
ARC ¿

} Reference Range: 20 X 109/L - 115 X 109/L

4. Corrected Reticulocyte Count

} is also known as Reticulocyte Index or Hematocrit Correction
} used to correct the reticulocyte counts in patients with
anemia
} Formula:
Patient Hematocrit ( %∨L/ L )
CRC=
45 %(0.45 L/ L)

} Reference Range:
o Hct of 35% (0.35L/L) expected to have 2% - 3%
o Hct of 25% (0.25L/L) expected to have 3% - 5%
o NV: same w/ the stated NV of Retics count – 0.5% - 2%

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 5. Corrected Reticulocyte Count
} also known as Shift Correction
} a general indicator of the rate of effective erythropoiesis in patients w/
anemia
} Formula:
CRC
RPI=
Maturation Time

Maturation Time Correction Factor:

Patient’s Hematocrit Correction Factor
Value (%) (Maturation Time,
Days)
40–45 1
35–39 1.5
25–34 2
15–24 2.5
<15 3

} Reference Range:
o >3 – indicates adequate bone marrow
o <2 – indicates inadequate erythropoietic response
} Clinical Significance:
Elevated RPI Decreased RPI
1. Chronic Hemolysis 1. BM failure
2. Recent Hemorrhage - Aplastic Anemia
3. Response to 2. Ineffective
Therapy Erythropoiesis
- Megaloblastic
Anemia
*Vit. B
12
*Folate
deficiency

Methods of Reticulocyte Count

1. Wet Method (Water)
a. NEW Methylene Blue Method (NMB)
b. Cook, Meyer, Tureen Method (BCB)
c. Seiverd’sMethod (MB or BCB)
2. Dry Method (Alcoholic)
a. Schilling’s Rapid Method (BCB)
b. Sabin’s Method (NR – Neutral Red & JG – Janus Green)
c. Seiverd’s (MB or BCB)
d. Osgood- Wilhelm (MB)

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 Note: NMB and BCB
} bind, neutralize & cross link RNA
} these stains cause the ribosomal & residual RNA to coprecipitate
with the few remaining mitochondria & ferritin in young
erythrocytes to form reticulum (filaments) & microscopically
visible dark – blue clusters

Clinical Significance of Reticulocyte Count

Increased Reticulocyte Count
Indicates Hemolysis
Physiologic Increase at birth, menstruation producing red blood cells (decreased
& pregnancy
hemolytic anemias,
individuals with IDA receiving iron
therapy,
thalassemia,
sideroblastic anemia,
high elevation, meds like LEVODOPA,
malarial meds, CORTICOTROPIN, &
fever-reducing meds and in
acute and chronic blood loss
Decreased Reticulocyte Count
aplastic anemia,
in conditions in which bone marrow is not
erythropoiesis),
acute benzol poisoning,
pernicious anemia &
chemotherapeutic radiation – induced
hypoproliferation (Azathioprine,
Chloramphenicol, Dactinomycin,
Methotrexate & other chemotherapy meds

Sources of Error in Reticulocyte Count

1. Refractile appearance ( bodies) of RBCs are confused as retics
2. RBC inclusions are mistaken as retics

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