I have stained PBL after ficol with CD3-biotin,CD19-biotin and CD16-biotin followed by Avidin-Cascade Blue from Molecular Probes. I have used about 1ug/sample of Avidin-Cascade Blue and have found a non-specific very bright stained population in all three samples. My impression is that Cascade Blue Dye is binding non-specifically for some reason. I am exciting dye with about 100MW of 407nm Kryptonn Laser line and collecting emission with a 450nm/65 Band Pass filter. If anyone has an idea what I am seeing with this bright non-specific staining I would appreciate comment. Thanks. David M. Dombkowski dombkowski@helix.mgh.harvard.edu Flow Cytometry-Pathology-CNY rm7017 149 13th Street Massachusetts General Hospital-East Boston, MA 02129 Tel. (617)726-1683 Fax.(617)724-3164
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