Search the CONFOCAL archive at
http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocalHi Fredrik,
This isn't a quantitative answer, but the light that the sample sees
during excitation, even with substantial ND filtering, etc. has to be
orders of magnitude more intense than ordinary room light.
Maybe someone has done the equivalent of what we do to test safelights
for film in a darkroom - make a set of slides and start with all of them
in the light but one (T=0, minimal necessary handling exposure) and then
dark-store one from the set at 5m, 30m, 1hr, 2hr. etc. Quantitatively
image the set at the end taking good care not to bleach in the process
of focusing. See if there is a difference. It is going to depend on the
quality and intensity of "room light" and how much fluorescence, window
light. But it will tell you, for your conditions, how much it matters.
Please let us know the results!
Dale
Fredrik Wermeling wrote:
> Search the CONFOCAL archive at
>
http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal>
> Hi,
>
> A trivial question that I’ve thought about is how much fluorophore
> bleaching that could be attributed to working with it under regular
> light (light bulbs etc). Some people tend to turn off all lights as soon
> as they think about a fluorophore. I seldom turn off the lights and have
> never had a problem with it. So my question is how much bleaching the
> light from a regular light source can cause. I do realize that it
> depends on how long the exposure time is and which fluorophore we’re
> talking about so to make it easy lets say the time it takes to mix, add
> and wash the samples (not the incubation step) and the fluorophores FITC
> and Alexa-488.
>
> /fredrik
>
>
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