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Physiological Practical 3

Cellular Physiology: Blood: Practical Haematology


The aim of this practical is to
1. Use blood typing techniques to determine the ABO and Rh blood group of
blood samples
2. Use a haemocytometer to determine the red blood count in a sample of
blood
3. Obtain a measurement of Packed Cell Volume (PVC) from a sample of
fractionated blood and use the PVC to calculate the Mean Cell Volume of the
red cells. (Reference book)
Experiment 1: Blood Typing
Method: The method for this experiment can be found on page 72 of the
Human Biology lab manual
Results: The results were compiled from
different students (including myself) in the laboratory on Monday 12 May 2014

Sample

Anti-A

Anti-B

Anti-D

D8054468

Blood
Type
O+

D4963181

B+

D8148725

O+

D4913662

A-

D8052526

B-

D9140495

A+

D8053303

O+

D9294804

O+

D9052526

B-

D9205119

O-

D9148445

A+

Preliminary notes for experiment 1:


- The blood samples that showed agglutination in the presence of Anti-A
antiserum had A antigens on their red blood cells and the blood was ABO
blood type A.
- The blood samples that showed agglutination in the presence of Anti-b
antiserum had B antigens on their red blood cells and the blood was ABO
blood type B.
- The blood samples which showed no agglutination to Anti-A and Anti-B
antiserum had no A or B antigens present on their red blood cells and the
blood was ABO blood type O.

None of the blood samples agglutinated in the presence of both Anti-A and
Anti-B (did not have both A and B antigens present) therefore none of the
samples were ABO blood type AB.
The blood samples that agglutinated in the presence of Anti-D tested
positive for the presence of the Rhesus factor, thus were Rh blood typed
Rh+. The samples that did not agglutinate were type Rh-.

Experiment 2: Red Cell Count


Method: For this experiment the procedure can be found on page 74 of the
Human Biology lab manual
Results: the more squares counted the greater the accuracy so I counted 96
instead of the minimum 80 squares.
Square (1
square
contains 16
small
squares)

Cell
Cou
nt

122

88

92

120

113

120

Total

655

Calculations:
Red Cell Count =

number of cells counted


6
dilution 10
Number of small squares volume of small square ( L )

655
200 106
Red Cell Count =
1
92(
)
4000
Red Cell Count =

5.696 1012 cells per litre

Preliminary notes for Experiment 2:


- Some of the red cells had lysed and had lost their ovoid shape.
- The red cell count obtained is within the standard deviation of the average

RCC for both men and women (according to Human Biology Lab Manual
page 77) so the results are credible.

Experiment 3: Packed Cell Volume - PCV


Method: The method for this experiment can be found on page 76 of the
Human Biology lab manual
Results:
Packed Cell Volume = 0.38 L/L
Calculations: The blood used for the calculation of PCV and the blood
used for the calculation of RCC is from the same person, so it is possible to
use the two results to calculate MCV
MCV = PCV/RCC
MCV = 0.38 /
MCV = 6.67
MCV = 66.7

5.696 1012
1014 L/L
fL

Preliminary notes for Procedure 3:


- When blood is spun in a centrifuge, the different components split into 3
separate phases; a plasma phase and haemotocrit phase separated by a
buffy coat.

Plasma makes approx. 55% of the total blood volume and contains plasma
proteins.
The buffy coat, the phase between the plasma and the packed cell volume,
is composed of platelets and white blood cells.
The haematocrit, also called the packed cell volume, contains the red blood
cells. Generally the PCV makes up 45 % of the total blood volume and the
results fall within the standard deviation of the average PCV.
The MCV calculated falls within the standard deviation of the average adult
MCV.

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