We are interested in detecting BrdU incorporation in GFP+ cells as an assay
of cell proliferation. The approach we want to take is to transiently
transfect cells with gene of interest and GFP. After 48hrs, we label the
cells in BrdU-containing media, and measure the number of cells that are
both GFP+ and BrdU+ compared to GFP+ BrdU- cells (using immunofluorescence).
This has been reported before (Biotechniques, 1997, p. 88), but the paper
lacks details on how this was done. Specifically,
- how is the GFP preserved when labelling the BrdU?
- what fixation methods preserve the GFP fluorescence?
Does anyone have any advice regarding these? Thanks in advance to any help
you can provide. Either post or email haqr at oci.utoronto.ca, and I'll
summarize for the newsgroup.