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Hi Gareth:<br>
<br>
I have not seen an answer to your question on Histonet so .........
Osmium (1% aqueous or less) will enhance the DAB reaction product. If
your (brain) sections have not been defatted/delipidized osmium will
also stain the lipids (myelin) in the brain. This may actually reduce
the contrast of the DAB-osmium reaction product depending on where it
is. Osmium will also make the sections less flexible so they should be
mounted on slides before exposure. Osmium is expensive and must be used
under a hood ("fume cupboard") as its vapors will fix your corneal and
nasal epithelium. It is also requires some care in its disposal. There
are several less dangerous/expensive ways to enhance the DAB reaction
product, including nickel or nickel+cobalt ions in the DAB-peroxide
incubation medium is popular. While nckel and cobalt are certainly not
benign they don't have the fume problems that osmium does and they are
less expensive. I can send you a nickel+cobalt recipe if you like. In
my experience DAB should be 'enhanced' with something if you want the
intensity of the reaction product to be permant. Also, use DPX as a
mounting media.<br>
<br>
Geoff<br>
<br>
Davis, Gareth wrote:<br>
<blockquote type="cite"
cite="mid082C721AF78DB34983E8BA2CD085462105C64B@mailbe07">
<meta http-equiv="Content-Type" content="text/html; ">
<div>I plan to do a immunohistochemical stain on mouse brain tissue
with c-fos. Some protocols I've read state that you should osmicate
the sections, after mounting, to enhance the stain. I've never stained
with c-fos, but I have been told it is very dark on it's own. Does
anyone know if it will be necessary to osmicate. Of course, I guess I
could just try and see, but any information on the staining process
itself would be helpful.</div>
<div>Thanks,</div>
<div>Ms. Gareth Davis</div>
<div>Vanderbilt University Medical Center</div>
<div> </div>
<div> </div>
<div><font face="System" color="#800080" size="2">Ms. Gareth B. Davis</font></div>
<div><font face="System" color="#800080" size="2"><strong>Research
Assistant II</strong></font></div>
<div><font face="System" color="#800080" size="2">Neuro-magnetics
Division of </font></div>
<div><font face="System" color="#800080" size="2">the Department of
Neurology</font></div>
<div><font face="System" color="#800080" size="2">Vanderbilt
University Medical Center</font></div>
<div><font face="System" color="#800080" size="2">615-936-3318</font></div>
<div> </div>
<div> </div>
</blockquote>
<br>
<pre class="moz-signature" cols="72">--
--
**********************************************
Geoff McAuliffe, Ph.D.
Neuroscience and Cell Biology
Robert Wood Johnson Medical School
675 Hoes Lane, Piscataway, NJ 08854
voice: (732)-235-4583; fax: -4029
<a class="moz-txt-link-abbreviated" href="mailto:mcauliff@umdnj.edu">mcauliff@umdnj.edu</a>
**********************************************</pre>
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