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2-Washing and Preparation of Red Cell Suspension

The document provides instructions for washing red blood cells and making a cell suspension. Red cells are washed 3 times by centrifuging, removing the supernatant, and resuspending the cell button in saline to remove proteins and fibrinogen that could cause non-specific agglutination. A 2-5% cell suspension is made by resuspending the final washed cell button in double the volume of saline. Washed red cells in the optimal concentration aid in detecting weak antibodies.

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Bromance H.
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2K views

2-Washing and Preparation of Red Cell Suspension

The document provides instructions for washing red blood cells and making a cell suspension. Red cells are washed 3 times by centrifuging, removing the supernatant, and resuspending the cell button in saline to remove proteins and fibrinogen that could cause non-specific agglutination. A 2-5% cell suspension is made by resuspending the final washed cell button in double the volume of saline. Washed red cells in the optimal concentration aid in detecting weak antibodies.

Uploaded by

Bromance H.
Copyright
© © All Rights Reserved
We take content rights seriously. If you suspect this is your content, claim it here.
Available Formats
Download as PDF, TXT or read online on Scribd
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Practical Session NO (2)

Done by:
Miss Nada Fallatah 2016
Red cells should to be washed to remove
serum or plasma which may contain:
1- proteins: that interfere with testing, causing
non-specific agglutination or rouleaux
formation.
2-fibrinogen: which may cause small clots.

Preparation of a 2-5% cell suspension


provides cells in an optimum concentration
to detect weak antibodies.
 Normal Ionic Strength Saline (NISS):
Phosphate buffer saline is suitable for
agglutination tests in tubes.

 Low Ionic Strength Saline (LISS): is used


to increase the binding of antibody to
antigen.
1. Sample:
 Anti-coagulated patient samples (EDTA).

 Donor blood taken from a sealed segment


of the donor unit.
2. Reagents, Equipment, and Supplies:

 Indeliblemarking pen.
 Glass test tubes.
 Disposal pipettes.
 Wash bottle filled with physiologic saline.
 Test tube stand.
 Centrifuge machine.
A- Washing:
1. Place 1 to 3 drops of blood to
new-labeled tube.
2. Fill the tube 3/4 full of isotonic
saline. Drops of blood
3. Centrifuge for 5 minutes at high with ¾ saline
speed (3400 RPM)
4. Carefully remove and discard
the supernatant
5. Shake the tube to re-suspend
cell button before washing the
cells again. Cell button in tube
NOTE: The red cell washing will
depend on the procedure being
done as to how many washings
are going to be done (usually
three times).

B- Making 3% cell suspension:


After the final wash, shake the
tube to completely re-suspend
the cells and add saline double
the amount of the red cell to get
2-3% cell suspension
a final concentration of
approximately 3%.
 To prevent contamination, do not touch tubes
with the crest of the saline bottle.
 Ifa sample from a donor unit is to be washed,
use a lancet to make a little fix in one of the
segments attached to the donor unit. Squeeze
a drop of red cells in the appropriately labeled
tube. Avoid applying excess pressure or the
cells will hemolysis.
 Re-suspend the cell button thoroughly
between washes before adding more saline to
ensure complete washing.
Red cell should be washed and suspended
in saline just before use to aid in increasing
the rate of antibody binding ratio as well as
unwanted antigens are easily taken out
from the red cell surface.

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