[Histonet] beta galactosidase nuclear counterstain

Shaumik Adhya shaumik.adhya <@t> ucl.ac.uk
Fri Apr 23 04:46:59 CDT 2004



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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