I find I get cleaner differentiation by using 1 ml HCl per liter of distilled water instead of the acetic acid solution.  2 minutes differentiation usually works well, but it can be used up to 5 minutes if necessary.

> ----------
> From: 	histonet-bounces@lists.utsouthwestern.edu on behalf of Liz Chlipala
> Sent: 	Wednesday, June 20, 2007 5:42 PM
> To: 	histonet@lists.utsouthwestern.edu
> Subject: 	[Histonet] picro sirius red
> 
> Hello everyone
>  
> We are running some picro sirius red for image analysis and were running into a bit of a problem.  We are staining rat heart, the collagen staining is quite nice but some of the muscle is staining a tad bit orangish rather than a really nice yellow.  Which is not a problem when you look under the microscope but once the images are captured for analysis thats were we run into problems.   We have tried making up fresh staining solution, etc.  differenciating more in the acetic acid but the muscle is stil a bit orange in color.  Any help would be appreciated.  
>  
> Thanks in advance
>  
> Liz
>  
> Elizabeth A. Chlipala, BS, HTL(ASCP)QIHC
> Manager
> Premier Laboratory, LLC
> P.O. Box 18592
> Boulder, CO 80308
> phone (303) 735-5001
> fax (303) 735-3540
> HYPERLINK "mailto:liz@premierlab.com"liz@premierlab.com
> HYPERLINK "http://www.premierlab.com/"www.premierlab.com
>  
> Ship to Address:
>  
> Premier Laboratory, LLC
> University of Colorado at Boulder
> MCDB, Room A3B40
> Boulder, CO 80309
>  
> 
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