With regard to Gene's ??, the following. Fig 1 in the paper clearly shows that the bimane and Ho42 spectra overlap alomst completely, so your plan to use a 440LP DC and a 450/50BP will fail completely. This was the reason we had to go the phase-resolved route. >From reading the literature we found several probes (mentioned in the article) that have been used for GSH/thiol analyses. However Hedley found that bimane was the most specific for GSH. If you are not that concerned about specifity you can probably use CMFDA and accept "some" co-staining of other -SH containing molecules. Indeed, providing a solution for the overlapping spectra was the central issue in the manuscript. Regards, Jan >Colleagues, >My question concerns the Phase-Resolved flow cytometry paper >by Jan Keij et al. in the january Cytometry. >I had thought people were routinely utilizing Hoechst/MCB to examine >glutathione and relative DNA content and had hoped to shortly do this >myself on our FACStar Plus utilizing UV excitation and collection >across a 440LP dichroic to a standard FITC filter fot the MCB and a >450/50 for the Hoechst. >Is monobromobimane much more specific for GSH and its overlapping >emission profile relative to Hoechst the central issue here? Gene/UCONN Health Jan F Keij Los Alamos National Laboratory LS-5, MS M888 Los Alamos, NM 87545 505-667-3526
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