Ann Atzberger writes- >Under the microscope one sees high (blueish) autofluorescence but also the >CFP signal i.e approx 60%pos. - not very bright but there. >So what I was thinking of doing is: > >staining the cells with CFDA, to interfere with the background >fluorescence, which isn't excited at 457nm. Right? Will it work?? Why not? If you're saying CFDA isn't excited at 457 nm, that's not right. It isn't excited optimally at 457 nm, but it's excited well enough to give you a lot of emission in your 515 nm detection band. Also, how good are the filter sets on your microscope; you shouldn't transmit much blue autofluorescence through a filter set made for blue excitation. -Howard
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