[Histonet] beta galactosidase nuclear counterstain
Gayle Callis
gcallis <@t> montana.edu
Mon Apr 26 10:37:44 CDT 2004
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
>
>Shaumik
>
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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 (lab with voice mail)
406 994-4303 (FAX)
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