[Histonet] lacZ staining in the liver
cbass <@t> bidmc.harvard.edu
Tue Feb 1 17:16:06 CST 2005
I was wondering if I could get some opinions (or protocols) on the best
way to see lacZ expression in the liver. I am injecting the liver
directly with a virus that should express lacZ. I would like to see
the efficiency of expression with this virus.
This is not a histology lab, nor am I very experienced so I have a
number of constraints about how I can best process this tissue.
I have easy access to a sliding microtome with a dry ice reservoir that
will section efficiently in the 20-40 micron range. Due to the large
number of animals, I would prefer to collect the tissue fresh and fix
by immersion in formalin. I would collect floating sections, and
probably stain the sections by floating them in the staining solution
(containing NP40 and SDS), mount them onto superfrost plus slides, and
counterstain with eosin, then coverslip with mount-quick.
Does this sound like it should work? Is there anything I should look
out for? For example, am I better off using some sort of subbed slide?
Is eosin the best counterstain?
If there are any other suggestions I would greatly appreciate it. I
used this methodology for lacZ staining in the brain and it seems to
work for me. However, I know very little about liver histology.
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