With 3-color we see significant reductions in staining intensity in the PE-Cy5 or PerCP-Cy5 signal (FL3) when there is also a strong PE signal. I assume this is steric hindrance. The antibody combinations of "kappa FITC, lambda PE and CD19 PE-Cy5 or PerCP-Cy5" and "CD10 FITC, CD34 PE and CD45 PE-Cy5 or PerCP-Cy5" show this reduction in the staining in FL3. We see this from different vendors and with a high % of samples which have the right antigen mix. Diluting the antibodies does not help eliminate this problem. Are others having this problem? Any practical suggestions as to how to avoid this? Thomas M. McHugh Technical Director, Clinical Laboratories UCSF Medical Center mchugh@pangloss.ucsf.edu
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