[Histonet] Reply on HELP!!! - Autofluorescence in the mouse brain

[Gayle Callis]

If you want, I have an excellent review of autofluorescence, the causes and 
what can or cannot be done to reduce the problem.  Although in conjunction 
with GFP, it is still pertinent to other fluorescent work.  I will be happy 
to attach it if you want, just let me know.

At 11:35 AM 11/12/2004, you wrote:
>Pawel,
>Sounds like you might be seeing lipofuschin granules, common 
>autofluorescent intracellular structures in brain.  To reduce 
>autofluroescence see
>Baschong W, Suetterlin R, Laeng RH.  Control of autofluorescence of 
>archival formaldehyde-fixed, paraffin-embedded tissue in confocal laser 
>scanning microscopy (CLSM).  J Histochem Cytochem. 2001 Dec;49(12):1565-72.
>They describe studies using ammonia-ethanol, borohydride, and
>sudan Black B alone or in combination, with a variety of fixation and 
>section prep methods compatible with immunolabeling.  You can find other 
>literature regarding the application of these reagents to autofluorescence 
>problems.  Expect some trial-and-error to figure out what works best for 
>your particular specimens.
>
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Gayle Callis
MT,HT,HTL(ASCP)
Research Histopathology Supervisor
Veterinary Molecular Biology
Montana State University - Bozeman
PO Box 173610
Bozeman MT 59717-3610
406 994-6367 (lab with voice mail)
406 994-4303 (FAX)