The effect of erythrocyte lysing reagents on enumeration of leukocyte subpopulations compared with a no?lyse?no?wash protocol

Introduction Standard protocols in flow cytometry (FCM) require lysis of erythrocytes, which may induce an unwanted loss leukocytes as bystander effect. Methods In the present study, we investigated influence 6 laboratory using 4 different lysing reagents, FACS® Lysing Solution (FacsL), QUICKLYSIS® (QuickL), IOTest® 3 (NH4Cl), VersaLyse® (VersaL), and with added fixative (VersaFix) on relative quantity leukocyte subsets identified by CD3, CD4, CD8, CD19, CD14, CD16, CD56, CD45, applying a no-lyse-no-wash (NoL) protocol reference. addition, compared efficiency red blood cell (RBC) lysis. Results Peripheral samples from 52 individuals were analyzed. NoL was suitable reference method, but led to less clear-cut gating lymphocyte monocyte populations due wider distribution light scatter. Best completeness RBC remaining erythrocytes below 10% achieved NH4Cl VersaL. We observed 11% monocytes after QuickL. Lymphocyte counts 19% lower FacsL. Cell within compartment rather similar between methods exception B-cell (?8%) higher NK-cell (+11%) VersaL good accordance method also mean values all methods. Conclusion Our data show that reagents tested lead specific deviations quantitation erythrocyte