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Lesson 6 RBC Counting

The document outlines the procedure for counting red blood cells (RBC) using a hemocytometer, detailing the materials, steps, and calculations involved. It includes reference values for normal RBC counts in different demographics and highlights variations due to physiological and pathological conditions. Additionally, it provides rules for accurate counting to ensure reliable results.

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lian.ericka14
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22 views2 pages

Lesson 6 RBC Counting

The document outlines the procedure for counting red blood cells (RBC) using a hemocytometer, detailing the materials, steps, and calculations involved. It includes reference values for normal RBC counts in different demographics and highlights variations due to physiological and pathological conditions. Additionally, it provides rules for accurate counting to ensure reliable results.

Uploaded by

lian.ericka14
Copyright
© © All Rights Reserved
We take content rights seriously. If you suspect this is your content, claim it here.
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HEMA 311 I 2024

HEMATOLOGY 1 OUR LADY OF FATIMA UNIVERSITY

LECTURE I LABORATORY BACHELOR OF SCIENCE IN MED LAB SCI


THIRD YEAR I 1ST SEMESTER TRANSCRIBED BY: J. HERSHEY REYES

WEEK 4: RBC COUNTING

RBC Counting squares


● The central square for RBC is subdivided into 25
smaller squares
● The distance between each counting surface and
● The red cell count is the number of red cells in 1 coverslip (depth) is 0.10 mm
cu.mm. ● The total volume is 9 cu mm
● Calculate the number of RBC per liter of each
● The RBC count is one of the tests that are used side of the hemocytometer
for the diagnosis of anemia and polycythemia

Materials and Equipment

● Anticoagulated blood (EDTA)


● RBC pipette
● Diluting fluid (Kayen’s solution)
● Tally counter
● Counting chamber
● Gauze pad
● Microscope
● Test tube

Procedure

1. Draw blood up to 0.5 mark using the RBC pipette Area used in the actual 1/ 5 sq mm
2. Wipe the outside walls of the pipette with a clean cell count
gauze with normal saline solution
3. Dip the pipette into diluting fluid, then aspirate the Area correction factor 5
diluting fluid into the pipette slowly until the
mixture reaches the 101 mark Depth of the counting 0.1 or 1/ 10 mm
4. Gently rotate the pipette to mix the diluting fluid chamber
and blood
5. Mix for 5 minutes Depth correction factor 10
6. Discard the first 3-4 drops of the diluted sample
7. Prepare the counting chamber Dilution 1:200 or 1/200
8. Charge both sides of the hemocytometer with a
drop of diluted sample and allow to stand for few Dilution correction factor 200
minutes
9. While keeping the hemocytometer in a horizontal
position, place it on the microscope stage RBC count= no. of cells counted x 5 x 10 x 200 or
10. Using HPO, count the red cells in the 5 “R” Multiply number of cells counted x 10,000
square of the central secondary square
Variation Technique
Microscopically:

● One large square is made up of nine 1-mm ● Polycythemia or erythremia- blood is drawn to
squares 0.3 mark of the RBC pipette and diluent up to 101,
● Each of WBC squares is divided further to 16 1:333

1
● Anemia- blood up to 1 mark and diluent to 101,
the dilution is 1:100

Reference Values

Normal Values:

● Female: 3.6 - 5.6 z 10 12/L


● Male: 4.2 - 6.0 x 10 12/L
● At birth: 5.0 - 6.5 x 10 12/L

Physiologic Variation Pathologic Variation

- Increased count - Increase


in dehydration erythrocyte count
- Increase count in in polycythemia
exercise - Increase in
- Newborn children pulmonary
have higher tuberculosis and
counts than pulmonary
adults fibrosis
- Women have - Increase in acute
lower counts than poisoning
male - Decrease in
- Individual living at anemia and after
higher altitudes hemorrhages
have higher
counts

Rules in Counting

1. Observe the boundary line of the square


2. Count the cells touching halfway in and halfway
out of the upper and left boundary lines
3. Never count cells touching halfway in and halfway
out of the lower and right boundary lines of the
square
4. Never count cells drying preparation
5. Never count cells in underfilled or overfilled
chamber

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