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Animal Physiology Lab 3

This document provides instructions for counting red blood cells (RBCs) using a hemocytometer. It describes diluting a blood sample, loading it into the hemocytometer, and counting RBCs in specific squares under a microscope. It then explains how to calculate the RBC concentration per cubic millimeter by factoring in the dilution and counting area. Normal RBC count ranges are provided for humans and some animals. Sources of error and clinical applications are also discussed.

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58 views15 pages

Animal Physiology Lab 3

This document provides instructions for counting red blood cells (RBCs) using a hemocytometer. It describes diluting a blood sample, loading it into the hemocytometer, and counting RBCs in specific squares under a microscope. It then explains how to calculate the RBC concentration per cubic millimeter by factoring in the dilution and counting area. Normal RBC count ranges are provided for humans and some animals. Sources of error and clinical applications are also discussed.

Uploaded by

ao868598
Copyright
© © All Rights Reserved
We take content rights seriously. If you suspect this is your content, claim it here.
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Download as PPT, PDF, TXT or read online on Scribd
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Subject: Practical Animal Physiology

4th STAGE.
Animal Production Department

COLLEGE OF AGRICULTURE

DUHOK UNIVERSITY

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The Total Red Blood Cell Count

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Aim: To count the total number of Erythrocytes (RBC) per cubic
millimeter (mm3)
Principle: A known volume of blood is diluted 200 times with a
known volume of an isotonic fluid a drop of diluted blood is then
spread over graduated scale of a known depth and the cells are then
counted in a known area.

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Apparatus:
Hemocytometer: consist of:-
A. A special pipette (with red bulb marked, 0.5-101)
B. Improved neubare slide (consist of large squares)
C. Diluting isotonic fluid (Hayem’s solution)
Consist of:
NaCl 0.5 g%
Na2SO4 2.5g%
HgCl2 0.25g%
In 100 ml distilled water
HgCl2 prevent fungus growth
NaCl + Na2SO4 prevent blood clotting and with distilled
water they diluted the blood sample.

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Procedure:
1. Clean and dry the pipette & counting chamber.
2. The cover slide placed on lateral bare across the middle of counting chamber.
3. Fill (capillary or venous )blood into red pipette at 0.5 mark.
4. Fill reagent add up into the pipette to 101 mark.
5. Hold the pipette horizontally and start rotating it between your thumb and index finger.
6. Discarded the first 3-4 drops.
7. Fill in the counting chamber nicely.
8. Allow RBC to set down for 2-3 min.
9. Count 5 red squares under microscope
10. Calculate the RBC concentration.

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RBC Count (cont.)
0.2 mm.

W W

R R
1 mm. R 0.2 mm.
R R

W W

High 0.1 mm.


3 mm. 9
RBC Count (cont.)
Top
Left

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RBC Count Calculation
1. Volume of 1 red square = 0.2 x 0.2 x 0.1 cu.mm.
= 0.004 cu.mm.
2. Volume of 5 red square = 0.004 x 5 cu.mm.
= 0.02 cu.mm.
3. In 0.02 cu.mm. the RBC count =N (counted No.)
4. In 1 cu.mm. the RBC count = N x 1 / 0.02
= N x 50
5. The dilution for RBC = 0.5 / 100
= 200
6. The final RBC count = N x 50 x 200
= 10,000 N (/cu.mm.)

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RBC Count (cont.)
Calculation

Red cell count = number of cells counted (N)


x volume factor (=50)
x dilution factor (=200)
= N x 10,000

Normal range = 3.8 - 6.0 x 106 / cu.mm.

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Source of Errors:
1. Bad sampling.
2. Prolonged use of tourniquet or excessive squeezing of finger.
3. Defect in pipetting, calculation.
Clinical Application:
Normal Ranges:
Men 4.5-6.5 million /mm3 with a mean value (5.5±1) million /mm3
Women 3.8-5.8 million /mm3 with a mean value (4.8±1) million /mm3
Infants (new borns) 4 -6 million /mm3
Children 3month 3.2 – 4.8 million /mm3
Children 3 -6 years 4.1 -5.5 million /mm3
Goat 13-14 million /mm3
Sheep 10-13 million /mm3
Rabbit 5.5-6.5 million /mm3

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If the number is higher than normal range is called Polycythemia, but if the
number is lower than normal range is called Anemia
Physiological Factors affect on RBC:
1. Hypoxia: when the O2 is too little in the atmosphere such as high
altitudes ( increase in RBC number).
2. Old age
3. Sex
4. Digestion
5. Pregnancy

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Thank You For Your Attention

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