[Histonet] Re: Autofluorescence/Imunofluorescence protocols rat kidney

[Gayle Callis]

I have a question:  what do you mean by "not effecting the signals from 
Alexa 488 and Alexa 546 too much?"  Do you actually see some reduction in 
these two fluorophores? What about the fluoresceinated based fluorophores, 
FITC, Rhodamines, Texas Red?



At 02:11 AM 5/26/2006, you wrote:
>Hello!
>
>I recently evaluated several easy methods to quench autofluoresence for 
>paraformaldehyde-fixed paraffin sections of bovine tissues.
>The only thing that worked (but worked beautifully!) was Sudan Black B (CI 
>26150). A 5-30 min incubation in a 0.1% solution in 70% ethanol, after the 
>whole immunofluorescence staining procedure, eliminated practically all 
>autofluorescence in any tissue, without affecting signals from alexa488 or 
>alexa546 too much. It's like magic! The only drawback was that it seems to 
>quench the DAPI counterstain.
>
>Just prepare the solution carefully, it won't solubilize very quickly. And 
>wash the slides with plenty of running water after the Sudan treatment.
>
>With best regards,
>Mikael
>
>PS In addition to several chemical treatments, I also tried the recently 
>published method of irradiating the sections with fluorescent tubes of 
>specific colors. Did not have any effect at all. Anyone know what I could 
>have been doing wrong?
>
>--
>////////////////////////////////////////////////////////////
>
>  Mikael Niku             URL: www.helsinki.fi/~mniku/
>  University of Helsinki  Dept. Basic Veterinary Sciences
>
>  - Mitäkö mieltä olen länsimaisesta sivistyksestä?
>  Minusta se olisi erinomainen 
> ajatus!
>                                          - Gandhi
>
>////////////////////////////////////////////////////////////
>
>
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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
406 994-4303 (FAX)