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LEARNING OBJECTIVES
INTRODUCTION
Numerous procedures that is done inside the laboratory requires demonstration of antigen
and antibody reactions in vitro; thus, the addition of indicator cells to the system is needed. The best
example is the testing for naturally occurring ABO antibodies in the plasma donors and recipients that
requires the utilization of such suspension. There are commercially available cells that are packed in
different combinations, but the most common is A1 and B combination.
Two to five percent red cell suspensions are the universally employed indicator cells displays such
reactions. These suspensions are prepared by using previously washed anticoagulated blood.
A 2-5% red cell suspension is used for the following tube examination procedures:
• ABO and Rh typing
• Direct antiglobulin test and auto control
• Donor unit compatibility (crossmatch)
• Red cell phenotyping
A. Materials
C. Procedure
WASHING
1. Pipette 3mL of whole blood into 10-mL conical tube.
2. Fill the tube with NSS until it reached the 10 mL.
3. Cover the tube with screw cap and gently mix by inversion.
4. Centrifuge for 5 minutes at 3,400 rpm
5. Decant by aspirating the supernatant using a pasteur pipette. Be sure that the packed red
cells are not disturbed.
6. Resuspend the cells with another volume of NSS until it reaches the same initial
measurement.
7. Repeat steps 3-5
Note: washing the cells is preferably done 3 times
8. Set wash red cells aside
4. Cover the prepared tube with Nescofilm and mix gently (if the suspension is correct this is the
formula)
Note: For best results red cell suspensions should be used for testing on the day of preparation.
Variations in red cell concentration can markedly affect the sensitivity of tests results. If red
cell suspensions are too concentrated, it may mask weak agglutination. When red cell suspensions
are too low in concentration, they become difficult to visualize, and in extreme cases, a weak
positive can fail to be detected.
Exercise 3
2. Why is Pasteur pipette used to deliver the WPRBC rather than the serological pipette?
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Using the formula given above complete the table in preparing red cell suspension with
desired concentration.
References:
-Cardona, C.C, Martin , G.L, and Garcia- Meim, R. (2016) Laboratory Manual in Blood
Banking 2nd Edition