[Histonet] b-galactosidase in lung

[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% 
acetone.
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
   kiernan <@t> uwo.ca
   http://publish.uwo.ca/~jkiernan/
______________________________________

t-kuzniar <@t> md.northwestern.edu 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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