Autofluorescence.

From: Oberdan Leo (oleo@dbm.ulb.ac.be)
Date: Thu May 31 2001 - 10:26:57 EST


Dear all,

I am trying to compare cell surface expression of TCR/CD3 structures on
naive versus activated T lymphocytes. I have a question related to the
interpretation of the data. What if two cell types express similar
levels of a given protein (as judged by the mean channel fluorescence
following staining with a directly labeled antibody), but display
different autofluorescence ?  My guess is that since autofluorescence is
at least an order of magnitude lower than the Ab-FITC signal, just
ignore it and conclude that both cells express similar levels of the
molecule. Am I right ? Or do I need to consider autofluorescence ? One
possibility would be to change the settings (PMT) so that
autofluorescence of both cells falls within the same range, but I do not
like this solution......... The problem is even worst when comparing T
cells to dendritic cells or macrophages ...

Thanks ,

Oberdan LEO




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