Re: [Histonet] beta galactosidase nuclear counterstain

From:Gayle Callis

Just air dry sections after final water rinse (after nuclear fast red
counterstain).  You can do this in front of a fan and up to 2 hours.  Mount
a permanent coverslip and avoid dehydration through alcohols, etc.  

At 10:46 AM 4/23/2004 +0100, you wrote:
>Hi Histonetters,
>I'm trying to stain myocardium for beta galactosidase (resulting in a blue 
>precipitate), and have been trying to use Nuclear Fast Red as a nuclear 
>counterstain - whilst this gives me reasonable nuclear staining, the process 
>of dehydrating and mounting makes me lose all the beta gal.  If I use an 
>aqueous mount such as gycergel, the Nuclear Fast Red dissolves out over the 
>course of a week, which is less than ideal.
>My protocol for dehydrating and mounting is:
>10 quick dips in 70% ethanol
>2 min in 100% ethanol
>5 min in 100% ethanol
>5 min in Histoclear
>2 min in Histoclear
>DPX to coverslip with.
>I'm wondering what people use as a nuclear counterstain for beta-gal, and if 
>you had any tips or advice.  Thanks
>Histonet mailing list
Gayle Callis
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)

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