Manual red blood cells count

Red Blood Cell

• Introduction:
• Non-nucleated formed elements of blood which plays a vital role
in transport of respiratory gases.
• Color – red, presence of pigment hemoglobin.
• Shape-Biconcave disc.
• Life span – 120 days.
 Functions of RBCs
• Transport of oxygen from lungs to tissue. 97%-Oxyhemoglobin.
• Transport of carbon dioxide from tissue to lungs.
• Buffering action. Regulates acid base.
• Blood group determination. ABO and Rh antigens.
 What Is a Red Blood Cell Count?
• The test is important because RBCs contain hemoglobin, which
carries oxygen to your body’s tissues.
• The number of RBCs you have can affect how much oxygen your
tissues receive. Your tissues need oxygen to function effectively.
• However, hemoglobin concentration and hematocrit
determination can be used.
 Reagents, supplies and equipment:
• Red blood cells diluting fluid (reagent):
– 0.9% Nacl.
• Sample:
– EDTA-anticoagulated blood
• Equipment:
1) Micro pipette.
2) Test tube.
3) Hemocytometer.
4) Coverslip.
5) Microscope.
6) Hand counter.
7) Lint-free wipe.
8) Alcohol pads.
 Procedure
1. Dilution of the blood:
• 1:200 dilution ( 10µL of blood + 1990 µL diluent).
2. Mix continuously for 2-3 minutes.
3. Load the cleaned hematocytometer.
4. Place the hematocytometer on the microscope stage.
5. Focus with x10 objective lens on the large central square.
6. This square is ruled into 25 small squares, each of which is
further divided into 16 smaller squares, of the 25 squares, only
the four corner squares, and one middle square are used to
count RBCs.
6. Switch to 40 objective lens, and start counting in the five
designated squares.
 Calculations
• The general formula for manual cell counts is as follows and can
be used to calculate any type of cell count:

Cells/ µL
1. Anemia( decreased number of Red blood cells).
2.  Polycythemia ( increased number of Red blood cells)
• Reference range:
• RBC, male 4.20–6.00 x1012/L
– 4.20–6.00 x109/mL
– 4.20–6.00 x106/µL
• RBC, female 3.80–5.20 x1012/L
– 3.80–5.20 x109/mL
– 3.80–5.20 x106/µL
• Normal Values:
• Newborn
• 4.1- 6.1x106/μl 0-1 day
• 4.36–5.96 x106/μl 2-4 days