physiology lab 1, wbc
DESCRIPTION
Physiology Lab 1, WBCTRANSCRIPT
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WBC manual count using hemocytometer
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Introduction
Blood represent about 8% of total body weight
It consist of 3 type of specialized cellular elements:
Erythrocytes RBCs Leukocytes WBCs Platelets (thrombocytes)
These cells suspended in complex liquid plasma
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Introduction Blood perform two major
function:1. Transport through the body
O2 & CO2 Food molecules (Glucose, lipid, aa) Wastes (ex. Urea) Hormones heat
2. Defense of the body against infections & other foreign materials, all WBCs participate in these defenses
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Introduction
All various types of blood cells are produced in the bone marrow
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Introduction
WBCs (leukocytes) Have nuclei Consist of:
lymphocytes with relatively clear cytoplasm
And 3 types of granulocytes whose cytoplasm contain granules
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Introduction
WBCs count is the count of leukocytes in a volume of blood
Expressed as WBCs/mm³
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Principle
The # of WBCs is very large, so its practical to dilute a sample with diluting solution (2% Glacial acetic acid with methylene blue)
This solution will lyses cytoplasmic membrane, and leave the nuclei of WBCs.
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Material
1. Blood sample (EDTA anticoagulated blood or capillary blood)
2. WBCs diluting pipette3. Diluting solution (2% AA with
methylene blue)4. Hemocytometer5. microscope
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The Hemacytometer
contains 2 Neubauer counting chamber
Each chamber contains: *4 WBC counting
squares *Each contains 16
squares
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The Hemacytometer
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The Hemacytometer
]0.25mm
]0.20mm
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Methodology
With a safety bulb draw blood up to 0.5 marks on WBCs pipette and complete to 11 with WBCs diluting solution.
Mix for 2-3 minute.
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Charge hemacytometer
Discard the first 4-5 drops
Place tip of the pipette at the edge central platform
Then let the hemacytometer to stand on the bench for 3-5 minute.
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Count and calculate:
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Count and calculate:
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Calculation
Number of cells/mm³= counted cells in 4 large square x diluting factor x volume correction factor.
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Calculation
The dilution factor= total volume/sample volume = 11-1/0.5 =20 Volume correction factor= Desired volume/counted volume 1 mm³/ counted volume
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Calculation
counted volume =The total volume of the 4 large squares== Volume x number of large squares= (width x length x depth )x 4= (1mm x 1mm x 1/10 mm) x 4
=0.4mm³ Volume correction factor= 1 mm³/ counted volume = 1 /0.4 = 2.5
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Calculation
Number of cells/mm³= counted cells in 4 large square x diluting factor x volume correction factor.
Number of cells/mm³= n x20 x 2.5 = n x 50
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Example:
If total # of WBCs in 4 squares is 120 Then the # of WBCs in 1mm³= 120 x 50 =6000
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Normal values of WBCs:
Newborn 9000-30000 cell/mm³ Adult 4000-11000 cell/mm³
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Significance of the test
Leukocytosis Occur as an indicator of body defense
against foreign materials (bacteria, parasites, toxins)
Metabolic disorders Chemical and drug poisoning Acute hemorrhage
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Significance of the test
Leukopenia Result from X-ray therapy Alcoholism Antibiotic therapy Typhoid infection Measls Infectious hepatitis TB And cirrhosis of the liver
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Sources of error
1. Flooding of chamber with excess sample
2. Failing to count all the cells in the squares or conversely including artifacts in the count.