Quantitative differential agglutination method using the Coulter Counter to measure survival of compatible but identifiable red blood cells.
The method described here using a centrifuge and Coulter Cell Counter for the quantitative differential agglutination of human red cells uses commercial anti-A and anti-B antisera for the ABO system, and for the Rh system a commercial anti-D serum, a low ionic strength solution and an anti-human IgG antiserum. We compared this Coulter Counter method with the Technicon AutoAnalyzer method which utilizes bromelin and polyvinyl pyrrolidone, and anti-A, anti-B and anti-CD antisera, and found this new method to be the simpler of the two. The nonagglutinable count with the Coulter Counter was 1.07% for A1 red cells, 2.26% for A2 red cells, 1.06% for B red cells, and 1.78% for Rh-positive red cells, results similar to those seen with the Technicon AutoAnalyzer. Results with the Coulter Counter method were consistently accurate whether the ACD red cells were studied on the day of collection, after 10 days of 4 degrees C storage, or after 4 degrees C storage for up to 6 days followed by cryopreservation with 40% (w/v) glycerol at-80 degrees C, thawing and washing. In this study, red cell samples obtained from recipients who had received compatible but identifiable donor red cells were frozen with 40% glycerol and stored at-80 degrees C for 10 months, thawed and washed. Survival measurements on these washed previously frozen red cells were similar to the values in liquid-stored red cells.