Physiology Labrotaroy

Red blood cell counting

Lab. 6

Dr. Iman AL-Nasrawy

Introduction:
A red blood cell count is a blood test that your doctor uses
to find out how many red blood cells (RBCs) you have in
your blood. It is also called an erythrocyte count .
This 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 are receiving. Your
tissues need oxygen to function effectively, This test
also evaluates the shape and the size of the red blood
cells. All of this information is then used to
determine the number of red blood cells per
microliter of blood .
Red blood cell count values vary according to the age
and the sex of a patient. The RBC count ranges from
4.2-5.0 million red blood cells per microliter of blood
for women and 4.6-6.0 million for men. A normal red
blood cell count for children is typically between 3.8
and 5.5 million red blood cells per volume .
This blood test is considered a very important indicator of a
patient’s health. A low red blood cell count might mean the
patient has anemia, acute or chronic blood loss,
malnutrition, chronic inflammation, or a number of
nutritional deficiencies including iron, copper, vitamin B-
12, or vitamin B-6. On the other hand, a higher than
average RBC count, called polycythemia, can be a sign of
congenital heart disease, pulmonary fibrosis, or renal
problems.
An increase of red blood cells can also happen naturally,
though. People who live at high altitudes tend to have a
higher-than-average RBC count, and smokers generally
have a higher number of red blood cells than non-
smokers. Drugs that can increase the RBC count include:
Gentamicin, Methyldopa while the Drugs that can
decrease the RBC count include :Chemotherapy drugs
,Chloramphenicol , Quinidine
Methods :
1- Manual method
2- Electronic method
Manual method :
Materials :
1-Capillary blood (lancet, alcohol)
2-Diluting fluid ( haymes solution in
which contain:
a- HgCl2 0.25 gm
b- Na2SO4 2.5 gm
c- NaCl 0.5 gm
d- Distilled water 100.0 ml
3--heamocytometer:
a- RBCs pipette b- rubber c- mouth piece
- Neubauers counting chamber with cover
slip ( two types:
a- Improved ( has groves and 25 middle
squares)
b- Ordinary ( without groves and 16 middle
squares)
- Light Microscope
Procedure :
1-The red blood cell pipette is filled with blood to the 0.5
mark in an identical manner and then filled to the 101 mark
with haymes solution. which prevents lysis and crenation
of erythrocytes. haymes solution also contains a fixative
which acts as a preservative and prevents agglutination of
cells if counting cannot be done within an hour.
2-Mixing is accomplished just as with the red cell pipette.
3-The counting chamber is conveniently located on the
microscope stage. Place the cover slip over the counting area
with the edge resting on the cover glass supports .
4-Shake the pipette again to insure adequate mixing. Expel the
fluid from the lower stem of the pipette and wipe the tip .
5-Place the tip of the pipette exactly at the junction of the
cover glass and the chamber. The capillarity of the space
between the cover glass and the chamber will immediately
pull fluid from the pipette to fill the chamber. If the fluid spills
into the moats or should bubbles form under the cover slip,
the chamber will have to be cleaned and the procedure
repeated.
calculation:
By using special formula we can multiply total
countable erythrocytes by 10000 (micro liter) or104
Total n. of RBC=N *10000
N=countable RBC in 80 squares.