Re: [Histonet] b-galactosidase in lung

From:John Kiernan

According to Takahashi et al J Histochem Cytochem
51(4):553-554 (2003) beta galactosidase staining
was strongest after fixation of cryosections
for 10 min to 8 hrs at room temperature in 100% 
There was weaker staining after 0.2% 
glutaraldehyde and almost no staining after
4% formaldehyde (from paraformaldehyde) - all
done for the same times & temperature. They
were looking at muscles from LacZ transgenic
rats, and they used the usual indigogenic
method, with a 30 minute incubation.

Could your detergent wash be doing something
nasty? It seems a very harsh thing to do
to sections of almost unfixed tissue, and
there's no obvious reason for it.
John A. Kiernan
Department of Anatomy and Cell Biology
The University of Western Ontario
London,   Canada   N6A 5C1
______________________________________ wrote:
> Hi,
> I'm having troubles finding my b-gal-expressing plasmid in the lung
> tissue. I do 5-10 um frozen sections of murine lungs, fix them for 10
> minutes in 0.2% glutaraldehyde, wash twice in a detergent wash, and
> incubate overnight in regular lacZ staining solution.
> I am sure my lacZ is there, because i can see it when I stain my lungs
> macroscopically.
> Thanks
> Tom Kuzniar
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