Erthrocytes must be washed within 4 hours of draw.
Collection:
1.) Collect and process a minimum of 2 mL of whole blood as follows:
A. Immediately centrifuge for 10 minutes at 1900 RPM
B. Discard the plasma and buffy coat layers.
C. Add a cold 0.9% saline solution to the erythrocytes (about 2 times the volume of erythrocytes).
D. Mix gently by inversion and centrifuge again for 10 minutes at 650 x g.
E. Remove and discard the saline.
F. Repeat the wash steps (C-E) 2 more times.
2.) After the final centrifugation, remove and discard the saline and a thin layer of the top cells.
Additional information: The relative centrifugal force (G-force) can be estimated by applying the following formula: g=11.18 x r (n/1000)2. Where: r=radius in centimeters and n=speed in RPM. The radius from the center of the rotation axis to the bottom or outermost portion of the test tube should be used. RCF is expressed relative to the force of the earth's gravity.