BLOOD PRACTICAL-1
Objectives
1. Methods used to measure the different
hematological values, and compare it with
the normal values.
2. The calculation of RBC indices, their
normal values and their importance in
diagnosis of different types of anemia.
3. Method used for determination of PCV
(Hct) value.
1. Obtain CBC (RBCs, WBC, &
platelets).
2. Determination of hemoglobin
concentration.
3. Determination of packed cell Microhematocrit
volume (PCV) hematocrit.
analyzer
4. Calculation of red blood cell
indices.
Coulter Counter
General Principle
 It counts and measures
the sizes of particles
(cells) by detecting and
measuring the changes
in electrical resistance
when a particle in a
conductive liquid pass
through a small
aperture.
An electrical current is passing This current is disturbed when a
through the aperture
cell passes through the aperture
The number of disturbances =
number of particles
The amount of disturbance =
volume of particles
Steps
1. Collect blood sample in anticoagulated glass
tube.
2. Diluted by device and it will count RBCs.
3. A lytic reagent will be added to blood sample.
4. Lysis of RBCs for WBC count and Hb
measurement.
Normal Values
RBC
WBC
HB
Platelet
Male
Female
Average
4.5-6.5
x106/l
4 – 11
x103 /l
13-18
g/dl
150400x103
/l
3.8-5.8
x106/l
4 – 11
x103 /l
11.5-16.5
g/dl
150400x103
/l
4.7–6.5
x106/l
4 – 11
x103 /l
13 –18
g/dl
150400x103
/l
Clinical application
1.
2.
3.
4.
5.
6.
 RBC = aneamia
 RBC = polycythemia
 WBC = leucopenia
 WBC = leucocytosis
 Platelets = thrombocytopenia
 Platelets = thrombocytosis
Measurement of Heamatocrit
Collect blood sample in
heparinized capillary
tube
Centrifuge
Read by heamatocrit reading scale
Normal value
Male
PCV % 40-54
Female
Average
35-47
35-54
Clinical Application
 Hct (PCV):



 RBC (polycythemia) ;
 plasma volume (hemo-concentration)
 Hct (PCV):



 RBC (anemia),
 plasma volume (hem-dilution)
Calculation of RBC Indices
 Mean corpuscular volume (MCV)=
 Average RBC volume.
 MCV= Hct X 10/RBC
 Normal= 80-100 fl
(fimtolitre ?)
 Mean corpuscular heamoglobin (MCH)=
 Average weight of Hb in RBC.
 MCH= Hb X 10/RBC
 Normal= 27-31 pg
(picogram?)
Calculation of RBC indices
 Mean corpuscular heamoglobin
concentration (MCHC)=
 Concentration of Hb in 100ml of RBCs.
 MCHC= Hb X 100/Hct
 Normal= 32-36 g/dl
Clinical Application
 Helps in the diagnosis of aneamia.