osmiumtetroxide enhancement of immunocytochem.
aray at emory.edu
Fri Aug 18 22:37:59 EST 1995
In article <40vpdo$m22 at agate.berkeley.edu>, <Noone at uclink.berkeley.edu> writes:
>>>I have done immuno against parvalbumin in brain tissue and staining is
>>>but weak. I've read that one can use osmium tetooxide to enhance
>>>of the precipitate from the DAB reaction. However, I need specific
>>>instructions about how to use the osmium to do this. If you can help me
>>>post in the
>>>news group or email me at kopl that is kopello at eden.rutgers.edu.
>>hi Michael, I can't provide the method but I thought you could do with
>>some OH & S stuff. Osmium tetroxide is mega- dangerous. It is both a strong
>>poison & mutagenic.
>I don't recommend using Osmium tetroxide to enhance DAB staining, unless you
want to investigate your tissue under the electron microscope.
We get excellent results with parvalbumin - fibers, cells, sometimes even
terminal fields. We use Ni ammonium sulfate, but don't have to to get good
staining. Are you sure you're using a high enough concentration of primary or
secondary (or tertiary if present) antibody? The other thing to wonder is if
you are using a long enough incubation in the DAB (or even in the Ab's). It
doesn't take much of a change in concentration or incubation to make a large
change in your labeling. Good Luck.
Dept of Neuroscience
aray at emory.ed
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