Light scatter based lymphocyte gate--helpful tool or source of error?
During flow cytometric analysis of 29 samples of human peripheral blood mononuclear cells, we observed that a light scatter-based lymphocyte gate would fail to cover all TCR (CD3)-expressing cells. A mean of 7.9 +/- 5.3% of all CD3+ cells was found outside the gate. In addition, this shifted cell subset showed an altered CD4/CD8 ratio (8.1 +/- 5.0% of all CD4+ and 13.7 +/- 5.8% of all CD8+ cells). The use of standard light scatter-based lymphocyte gates in acquisition and/or analysis of flow cytometry (FCM) data may thus lead to erroneous results, particularly if T cells are to be studied.
Krömer
E
E
Ludwig Boltzmann Institute of Leukemia Research and Hematology, Vienna, Austria.
Grossmüller
F
F
D016477:Artifacts; D002469:Cell Separation / Q000379:methods*; D005434:Flow Cytometry / Q000379:methods*; D005455:Fluorescent Antibody Technique; D006801:Humans; D016130:Immunophenotyping / Q000379:methods; D008027:Light; D012542:Scattering, Radiation; D016176:T-Lymphocyte Subsets / Q000166:cytology*