Accurate counting of low numbers of leukocytes. Use of flow cytometry and a manual low-count chamber.

Abstract

The leukocyte depletion that current filters make possible in erythrocyte and platelet preparations can result in leukocyte concentrations too low to be counted accurately by automated and standard manual methods. A recently described method for counting low numbers (1 to 10 x 10(6) per L) of leukocytes by flow cytometry and the use of a manual low-count chamber on 25 venous samples serially diluted to 1:1,000 were evaluated. The results show that both methods for counting leukocytes can reliably determine counts of 1 to 10 x 10(6) leukocytes/liter (a three-order of magnitude reduction from venous blood) in terms of order of magnitude but lack accuracy for specific measurement. The flow cytometric method is more expensive, less readily available, and suffers from greater sample variability. Use of a low-count chamber is a superior technique to evaluate and maintain quality control of methods for leukocyte depletion, resulting in a final leukocyte concentration of 1 to 10 cells per microliter.