Name: ___________________________________________ Date: __________________

Group: __________________ Score: _________________

RED BLOOD CELL COUNTING

EXPERIMENT 1
To achieve this unit a learner must:
Demonstrate the proper use of RBC pipette.
OUTCOMES

Perform the proper charging of blood into the hemacytometer.

UNIT

Focus the proper counting squares for RBC.

Perform RBC counting properly and accurately.
Calculate the RBC count accurately.

TEACHING AND LEARNING ACTIVITIES

Laboratory Experimentation

The students will demonstrate the proper procedures of red blood cell counting. The students
will answer the guide questions after the experimentation.

The red cell count is the number of red cells in 1 cu.mm. The RBC count is one of the tests that
are used for the diagnosis of anemia and polycythemia or erythrocytosis and leukemia. The normal adult
counts are as follows:

Males: 4.5-5.9 x 1012/L

Females: 4.0-5.2 x 1012/L

Glossary of Terms

Anemia – is a condition characterized by abnormally low levels of healthy red blood cells or hemoglobin
(the component of red blood cells that delivers oxygen to tissue throughout the body)

Erythrocytosis – an increase in the number of circulating erythrocytes of more than two standard
deviations above the mean normal, usually occurring in secondary hypoxia.

Leukemia – any of various acute or chronic neoplastic diseases of the bone marrow in which
unrestrained proliferation of white blood cells occurs, usually accompanied by anemia, impaired
blood clotting and enlargement of the lymph nodes, liver and spleen.
Polycythemia – an increase in the number of red blood cells.

ANALYTICAL PHASE
PRE-ANALYTICAL PHASE
Materials and Instruments
Anticoagulated blood (EDTA)
RBC pipet
Diluting fluid (Hayem’s or Gower’s solution)
Tally counter
Counting chamber
Clean gauze
Test tubes
Microscope

Procedure

1. Draw blood up to 0.5 mark using the RBC pipette.

NOTE: If the blood was drawn too far above the 0.5 mark, the procedure should be
repeated using a new pipette, excess blood causes an inaccurate result.
2. Wipe the outside walls of the pipette with clean gauze.
NOTE: Do not allow capillary attraction to draw fluid from the tip onto the gauze. Gauze
tends to absorb the liquid portion of the blood, causing an inaccurate result.
3. Dip the pipette into diluting fluid, and then draw the diluting fluid into the pipette
slowly, until the mixture reaches the 101 mark. Gently rotate the pipette to ensure a
proper amount of mixing. The dilution is 1:200
4. Remove the tubing from the pipette, and then mix it in a horizontal axis for 5 minutes.
5. Discard the first 3-4 drops of the diluted sample.
6. Charge both sides of the hemacytometer counting chamber with a drop of the diluted
sample and allow to stand for few minutes.
7. While keeping the hemacytometer in a horizontal position, place it on the microscope
stage.
8. Using HPO, count the red cells in the 5’R’ squares of the central secondary square (Fig.
1).
9. Calculate the number of RBC per liter of each side of the hemacytometer. Average the
results and report this number
FIG. 1 Close-up view of the counting chamber
of the Hemacytometer. The areas for RBC are
labeled as R, and the areas for WBC are
labeled as W.
FIG. 2 One square of hemacytometer,
indicating which cells to count. Cells touching
the top and left lines are counted, while cells
touching the bottom and right lines are not
counted. (Solid circle, cells counted; open

POST-ANALYTICAL PHASE

Data and Results

(3pts)
Patient’s Name:
________________________________________________________________________

Age and Gender: ________________ Date and Time of Collection:

__________________________

FORMULA:
Total count = Number of RBC counted x Dilution Factor x 106
mm2 counted x 0.1 mm

Computation Interpretation
(2pts)

Critical Thinking

1. Give at least 10 errors in red blood cell counting? (10 pts)

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 (STEININGER)
2. Why do you discard the first 3-4 drops before charging the sample in the counting chamber?
(2pts)

3. What are the rules in counting cells in the hemacytometer? (4pts)

4. Calculate for the RBC Count. Show your computation. (4pts)

RBC counted: 800
Squares counted: 5 RBC squares
Aspirated blood up to: 1 mark
RBC/cu