Preparation of 2 – 5% red

cell suspension
Laboratory Discussion 2
Introduction
Many procedure done in the laboratory require
demonstration of antigen and antibody reactions in vitro and
thus, addition of indicator cells to the system is needed. Two
to five percent (2-5%) red cell suspension (RCS) is the
universally employed indicator cells to display such reactions.
It is prepared by using an anticoagulated blood.
reagent
1. Normal Saline Solution (0.85-.9%) NaCl in a wash bottle
2. Anticoagulated Blood preferably blood with EDTA
Procedure:
Preparation of washed packed red blood cell
1. Pipette 2mL of blood intro a test tube using Pasteur pipette
2. Fill the tube half full with normal saline solution.
3. Cover the tube with nescofilm and mix by gentle inversion
4. Centrifuge for 1 minute at 3,400 rpm.
5. Remove the nescofilm and aspirate the supernatant using a Pasteur
pipette . This can also be done by discarding the supernatant as
quickly as possible in order not to disturb the cell bottom.
6. Resuspend the cells with another half-full volume of NSS. Cover
the tube with Nescolfilm and mix by gentle inversion
7. Centrifuge for 1 minutes at 3,400 rpm
Note: This process is called washing and is preferably done three
times
8. After the last washing, aspirate the supernatant using a Pasteur
pipette.
Procedure:
Taring an empty tube with a fixed volume of fluid
1. Using a 0.1mL serological pipette, aspirate distilled water up to the
highest mark.
2. Deliver exactly 0.1mL onto a clear identical test tube.
3. Mark the level
Note: This is now your 0.1mL tared tube. Set this aside.
PROCEDURE:
Preparation of red cell suspension of various concentration

1. Label 4 test tube as 2% RBC, 3% RBC and 5% RBC.

2. Using a Pasteur pipette, drop as much washed packed red blood
cell in a separate test tube until it reaches a volume equal to 0.1mL.
Compare that volume to a previously tared tube.
Note: Use only 1 Pasteur pipette in dropping the washed PRBC
3. Using a 2.0 mL or 5.0 mL serological pipette, deliver exactly the
following required volume of NSS
continuation
2% - 4.9mL NSS
3%- 3.2mL NSS
4%- 2.4mL NSS
5%- 1.9mL NSS
4. Cover with nescrofilm and mix gently
% Red Cell Suspension = amount of washed packed red blood cell x 100
total volume

Total volume= amount of washed packed red blood cell + amount of NSS to be added