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Dear Hai--

Are the mice alive or dead?  If alive, remove the hair with a chemical
depilatory (e.g. "Nair"--found at drugstores), if it's a problem.  The
superficial epidermis is highly autofluorescent and I don't know any way
around it except using a longer fluorophore, which I gather isn't an
option for you.  However, once you get into the dermis, that should be
largely gone.  You might try comparing results using 1-photon to those
using 2-photon; that might give you a better idea of the source of the
autofluorescence.  If 488 nm excitation doesn't provoke the same
autofluorescence (--I don't know how much you're seeing, but in my
experience there isn't THAT much), then you might want to consult with
an EGFP/2P person and see if you can change the 2P wavelength.

Good luck!

Martin Wessendorf

Qi, Hai (NIH/NIAID) wrote:
> We are trying to visualize the intradermal behavior of
> EGFP-expressing/// Leishmania/ parasites in mouse ears. A hurdle is the
> high background autofluorescence from keratinocytes, hairs, and hair
> follicles. Given that our samples were excited with a 2-photon laser at
> 800 nm, should I consider to tune the laser to a longer wavelength? Any
> additional suggestions on potential ways to reduce the autofluorescence
> are very much appreciated.


--
Martin Wessendorf, Ph.D.                   office: (612) 626-0145
Assoc Prof, Dept Neuroscience                 lab: (612) 624-2991
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