[Histonet] alternative to Bouin's in Schiff-methylene blue nucleic acid

[John Kiernan]

The hot Bouin will be working as a fixative for the bacteria in addition to causing partial hydrolysis of DNA and (probably) complete hydrolysis and dissolution of RNA. Bouin is the traditional fixative for making nuclear material (nucleoids) of bacteria visible. See Robinow C & Kellenberger E 1994 "The bacterial nucleoid revisited" Microbiological Reviews 58(2):211-232.
 
John Kiernan
Anatomy & Cell Biology
University of Western Ontario
London, Canada
= = =
----- Original Message -----
From: Carla M Conway <cmconway <@t> usgs.gov>
Date: Wednesday, January 19, 2011 15:17
Subject: [Histonet] alternative to Bouin's in Schiff-methylene blue nucleic acid
To: histonet <@t> lists.utsouthwestern.edu

> Hello,
> 
> A colleague is trying to determine the nucleic acid type of a 
> rickettsial 
> phage. Acridine orange with RNAse/DNAse treatment was not 
> successful. They 
> want to try a Schiff-methylene blue technique which requires 
> hydrolyzing 
> in Bouin's fluid (1 h at 60 C). Could anyone suggest another 
> protocol 
> which does not use picric acid (or suggest an alternative to 
> Bouin's )? 
> 
> Also, any advice for nucleic acid differential staining would be 
> appreciated.
> 
> Thanks very much, 
> 
> 
> 
> Carla Conway
> Histology Technician
> Western Fisheries Research Center, USGS
> 6505 N.E. 65th Street.
> Seattle, WA 98115-5016 USA
> Phone: 206-526-6282 ext. 242
> Fax: 206-526-6654
> E-mail: cmconway <@t> usgs.gov
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