[Histonet] RE: beta galactosidase nuclear counterstain
C.M. van der Loos
c.m.vanderloos <@t> amc.uva.nl
Mon Apr 26 04:37:33 CDT 2004
Hi,
Using X-gal as substrate and ferri/ferro-cyanide as chromogen for b-galactosidase activity you obtain a turquoise/blue reaction product that survives everything: alcohol, xylene, cooking in citrate, in situ procedure.....
Chris van der Loos, PhD
Dept. of Pathology
Academic Medical Center
Amsterdam - The Netherlands
---- Original Message -----
>From Shaumik Adhya <shaumik.adhya <@t> ucl.ac.uk>
Date Fri, 23 Apr 2004 10:46:59 +0100
To histonet <@t> lists.utsouthwestern.edu
Subject [Histonet] beta galactosidase nuclear counterstain
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
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