Re: [Histonet] Re: Autofluorescence/Imunofluorescence protocols rat kidney

From:Mikael Niku


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,

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:
  University of Helsinki  Dept. Basic Veterinary Sciences

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