Posted by
Alessandro Esposito on
URL: http://confocal-microscopy-list.275.s1.nabble.com/Bleaching-of-background-tp7578651p7578654.html
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Dear Sanjuan,
I can confirm that you may see autofluorescence in many media
formulations. We observe the same with Leibovitz media. I never
investigated the issue as it is not impacting our experiments, but there is
clearly some component of Leibovitz (I've seen identical results with
DMEM) that is autofluorescent. I would also add that I assume the source
of autofluorescence is a relativelty large molecule as it diffuse very slowly
to the photobleached field of view. I mention this because it has possible
impact on imaging. Very often, by the time you select a cell and find the
proper imaging parameters, autofluorescence is significantly bleached and
(with the appropriate filters) not visible on the acquired images. However,
for time lapse imaging, autofluorescence is recoverying because of the
long time between subsequent exposures and contributing to background.
I am not sure if there is anything better than Leibovitz in terms of
autofluorescence, perhaps other users have more experience.
Cheers,
Alessandro