Hematocrit determination

(Packed Cell Volume, PCV)

(Relative Corpuscular Volume)

Principle :
PCV determines the percentage (%) of red blood
cells (RBCs) in the whole blood .

Methods :
can be performed by two methods:
1. Microhematocrit method .
2. Macrohematocrit method .
Equipments :
1. Wintrobe macrohematocrit tube

2. Blood sample
(5 ml venous blood + 4 mg potassium oxalate + 6 mg
ammoniun oxalate).

3. Pasteur pipette : a special pipette with a long narrow tip

used in filling the wintrobe tube .

4. Centrifuge .
Wintrobe macrohematocrit tube:
- A glass tube of 10 cm length and 3 mm
diameter .
- - It carries to graduations (from 0 to 10) one
begins at the top of the tube and the other
begins at the bottom .
(pasteur pipette)
Procedure :
1. Mix the blood sample gently .

2. Fill two macrohematocrit tubes by using pasteur

pipette .

3. Put the two tubes in the centrifuge opposite to each

other (for balance) .

4. Centrifuge at 3000 rpm for 30 minutes .

5. Record the result .

(Wintrobe macrohematocrit tube after blood separation )
Equipments :
1. Capillary tubes , plain or heparinized (contains
heparin as an anticoagulant).

2. A blood sample .

3. Microhematocrit centrifuge .

4. Microhematocrit reader .

5. Clay sealant .
 (Microhematocrit tube)

(Microhematocrit reader )
Procedure:

1. Fill ¾ of the capillary tube with blood by capillary

attraction
2. Seal one end of the tube with clay .
3. Place the tube in the microhematocrit centrifuge
4. Centrifuge at a speed of 10000 to 12000 rpm for 5
minutes
5. Place the tube in the microhematocrit reader and
record the reading .
(Microhematocrit tube after blood separation)
The microhematocrite method is more proffered than
the macrohematocrite method due to:

1- It is more accurate due to less amount of plasma

trapped between the cells .
2- Needs less amount of blood
3- Needs less time.
Observation and Significance :
The blood within the tube will be separated into three
layers according to the specific gravity of the different
cell types :
1- Packed cell volume (PCV):
§ A red layer in the bottom of the tube containing RBCs
only .
§ The % of PCV can be calculated by the following
equation :

PCV(%)= Height of RBCs column (mm) x100

Height of blood column (mm)
§ the normal PCV values are :

Species PCV (%)

Horse 32-53
Cow 24-46
Sheep 27-45
Goat 22-38
Pig 32-50
Dog 37-55
Cat 24-45
Man 40-50
Human
Women 35-45
Newborn 40-65
Ø Lower PCV than normal may indicates anemia or
hemodilution .
Ø Higher PCV than normal indicates polycythemia,
hemoconcentration or dehydration (occurs in
sever burns).
Ø Hematocrit determination is useful in calculating
blood indices .
2- Buffy coat:
§ A gray layer at the middle of the tube containing
leukocytes and thrombocytes .
§ The normal buffy coat is about 0.5 – 1.2mm.
§ Importance: used for rough estimation of leukocytic
count;
- the first mm indicates presence of 10000 leukocytes /
cu.mm.
- while the second mm indicates presence 5000
leukocytes / cu.mm.

§ Lower buffy coat than 0.5 mm indicate leucopenia

§ Higher buffy coat than 1.5 mm indicate leukocytosis

3- Plasma :
§ A straw yellow layer on the top of the tube .
§ Normally plasma layer is about 55% of blood .
§ Lower values indicate polycythemia, hemoconcentration
or dehydration
§ Higher value indicate anemia or hemodilution
§ Abnormal colors of plasma may be observed :
ü Reddish plasma → haemolysis
ü Deep yellow → jaundice
ü Turbid plasma → lipemia