Topic: Red Cell Suspension
PRELIMS | Laboratory | Ma’am Geraldine Bediones Jungco
PREPARING AN EXACT 5% (and 2%) WASHED RED CELL SUSPENSION
1. Label test tubes as follows:
• Plasma/Patient’s identi,ication
• Washed cells/Patient’s identi,ication
• 5% Red blood cell suspension/patient’s identi,ication
• 2% Red blood cell suspension/ patient’s identi,ication
NOTE: Labeling is a critical step in any laboratory procedure and should be done carefully
2. Centrifuge the blood sample, if necessary. EDTA is the preferred anticoagulant during
preparation of red cell suspension.
3. Wearing gloves and goggles, use a transfer pipet and carefully remove the plasma to the
labeled tube.
NOTE: Be careful not to contaminate with red blood cells.
4. When most of the plasma has been removed, remove some red cells from the bottom of the
patient sample tube and place them in the tube labeled “Washed cells”.
NOTE: This tube should be no more than half full of cells.’
5. Add NSS from the wash bottle to the “Washed Cells” tube until the liquid level is about ½”
from the top.
NOTE: Be careful not to contaminate the tip of the wash bottle dispenser with blood.
6. Cover the top of the tube with paraUilm and invert to mix
7. Centrifuge with a balancer tube for 1 minute at 3400 rpm.
NOTE: Never open the lid of the centrifuge before the spinning action has come to a complete stop. The
spin time may vary on individual centrifuges. The student should check the calibration information for
the centrifuge in use.
8. Discard paraUilm into a biohazardous waste container.
9. Using a transfer/Pasteur pipet, carefully remove the supernatant and discard it into the
large beaker with disinfectant. Be certain not to splash any of the supernatant on top of the
laboratory table.
10. Repeat steps 5 through 9 two or three more times. This makes a total of three or four
washes.
11. Using the 5.0 ml pipet and a rubber aspirator, pipet 1.9 ml of saline from the small beaker
into the tube labeled “5% Red Cell Suspension”.
*(pipet 1.96ml of saline to tube labeled 2% RCS)
12. Using a 0.1 ml pipet and a rubber aspirator, add 0.1 ml of patient’s red cells
to the saline in the “5% Red Cell Suspension” tube.
*(pipet 0.04ml of patient red cells to saline in the 2% red cell suspension)
13. Cover with paraUilm and mix by several inversions. This is an exact 5% Red cell suspension.
14. Discard all biohazardous waste in a puncture proof biohazardous waste container.
Additional Notes:
• If other concentrations or volumes of red cell suspension are desired, the formula below can
be used to determine how much washed cells and NSS should be mixed together.
𝐴𝑚𝑜𝑢𝑛𝑡 𝑜𝑓 𝑊𝑎𝑠ℎ𝑒𝑑 𝑅𝐵𝐶𝑠 𝑥 100
% 𝑅𝑒𝑑 𝐶𝑒𝑙𝑙 𝑆𝑢𝑠𝑝𝑒𝑛𝑠𝑖𝑜𝑛 =
𝑇𝑜𝑡𝑎𝑙 𝑉𝑜𝑙𝑢𝑚𝑒
• Where: Total Volume = amount of washed packed RBCs + amount of NSS to be added
• The desired total volume can be set by the medical technologist.
• For instance, in order to prepare 5 ml of 2% red cell suspension, the amount of washed
packed RBC and amount of NSS to be added can be determined by manipulating the formulas
given above.
PREPARATION OF AN APPROXIMATE 5% WASHED RED CELL SUSPENSION
1. Label 2 test tubes as follows:
a. Washed cells/patient’s identi,ication
b. 5% Red cell suspension/patient’s identi,ication
2. Wearing gloves and goggles, perform steps 4 through 10 from the procedure for
preparing an exact 5% washed red cell suspension.
3. Using the tube labeled “5% red cell suspension”, Uill the tube half full with NSS from the wash
bottle.
4. Using a transfer/Pasteur pipet, add cells from the “washed cell tube” until the color and
concentration of cells in the tube is approximately the same as the exact red cell suspension
prepared in the previous procedure.
NOTE:
• As the cells are added, use the transfer/pasteur pipet to mix the solution by rinsing the pipet
with the saline/cell mixture in the tube.
• This transfer pipet may then be used to transfer the 5% cell mixture into the tubes to be
tested.
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