The red blood cell (RBC) count count are parts of the routine
complete blood count (CBC). RBCs are the most numerous of
the blood cells.
The RBC count approximates the number of circulating RBCs
and thus gives an indirect estimate of the blood’s oxygen-
carrying capacity (hemoglobin).
The RBC count is helpful in the diagnosis and treatment of
many diseases, especially anemia, a condition in which there
is a decrease below normal in the RBC count or hemoglobin
level.
1. with a bulb containing red bead as in haemocytometer or a
Sahli's pipette graduated to 20 µl, or any automatic pipette capable of
measuring 20µl volumes.
2. r with cover slip. It is a thick glass slide with H
shaped moats in it. Area between 2 limbs of H is 0.1 mm lower than area on
sides. When a cover slip is fixed across these limbs a depth of 0.1 mm is
provided in the centre. Above and below the horizontal moat is the ruled area.
Moat prevents mixing of two samples charged on either side.
3. . Prepared by dissolving 3.2 g of sodium citrate and 1.0
ml commercial formaldehyde solution in 100 ml distilled water.
4.
5.
6.
7.
1. Draw well mixed blood in RBC pipette up to mark 0.5. Care should be taken not to have air
bubbles in blood column (Blood can also be collected from a finger prick as well, Wipe the outer
side of the pipette clean.
2. Draw RBC diluting fluid up to mark 201 (1/200 dilution).
3. Gently rotate the pipette between thumb and forefinger to mix well.
4. Alternatively prepare 1/200 dilution of blood in diluent in a test tube by adding 20 µl of blood
to 3980 µl diluent.
5. Place the cover slip firmly on the Neubauer chamber. The sign of correct placing is that
diffraction rings are seen on either side.
6. Discard the first 4-5 drops from the RBC pipette before charging the chamber. Blood diluted in
test tube can be used as such after mixing.
7. Charge one side of the chamber by introducing a small drop of diluted blood at the edge of the
cover slip. The sample will move under the cover slip by capillary action.
8. Wait for two min to allow the cells to settle.
9. Count the cells using 40x objective in the central large doubly ruled square of the Neubauer
chamber. Select 5 small squares, four on corners and one in the centre for counting.
Cell counting areas: Cells in the four large corner squares (shown in blue) are counted for the white blood cell count; all
cells in the entire center square (yellow, circled) are counted for the platelet count; the four red corner squares and center
red square within the large center square (labeled a-e) are used for the red blood cell count
Loading the hemacytometer
Photomicrograph of red blood cells as they appear on a hemacytometer
Counting method: A sample hemacytometer square with triple boundary
lines showing cells to count; cells colored red should be counted and
green cells should not be counted
RBC Count/Liter = Cell Counted × DF × 106 / Area × Depth
For Instance, If Cell counted were 550 then,
Step 1- RBC Count/Liter = 550 × 200 × 106 / 0.2 × 0.1
Step 2- RBC Count/Liter = 110000 × 106 / 0.02
Step 3- RBC Count/Liter = 5500000× 106
Step 4- RBC Count/Liter = 5.5× 1012 / Liter
RBC Count = 5500000 µL or 5.5× 106 / µL or 5.5× 1012 / Liter