Double "mirror" lymphocyte population
Posted 22 July 2009 - 04:49 AM
I have problems with my cell culture/activation/staining protocol. I analyze the samples (cultured PBMCs or freshly isolated mucosal intestinal lamina propia lymphocytes) with a FACSCalibur cytometer. and frequently I find a double lymphocyte population: one is smaller (200-400 FSC) and the other bigger (400-600FSC). they have the same expression pattern (regarding CD8, CD45 and CD3). usually the bigger population has few CD4 compared with the small one.
Is that normal? Am I doing something wrong with my cells?
Posted 23 July 2009 - 05:27 PM
Posted 24 July 2009 - 01:41 AM
it sounds like "doublets" (two cells attached to each other). We used to see these in mouse lymphocytes samples
Posted 27 July 2009 - 05:52 PM
Posted 30 July 2009 - 05:10 AM
Thanks! How can I avoid or reduce that phenomenon? It normally increases with a long time culture.
Additionally to thoroughly mix the cells with the pipette, you could gate the doublets out for analysis.
Posted 25 August 2009 - 06:40 AM