I've heard of an approach for studying cell cycle control using GFP. GFP and
molecule X are co-transfected into a cell and GFP+ cells are scored for
incorporation of BrdU by florescence microscopy. How is this done? Doesn't
the fixation for BrdU staining destroy the GFP? Are cells simply marked for
GFP positivity and then fixed and stained, then realigned with the GFP
photo? If so, how is the alignment maintained exactly?
Also, has anyone tried sorting GFP+ cells which have already incorporated
BrdU? I'm just wondering if the exposure to the sorting laser destroys any
possibility of measuring BrdU positivity (as BrdU labeled cells must be kept
in the dark)?
haqr at oci.utoronto.ca