[Histonet] nissl after permount?

[Teresa Iglesias]

Thank you for the excellent information.
-Teresa

On Mon, Dec 5, 2011 at 1:03 AM, John Kiernan <jkiernan <@t> uwo.ca> wrote:

> The usual approach would be to do Nissl stains on nearby (ideally
> adjacent) sections to those immunostained for the immediate early gene
> products. If you have no unstained slides (from bad planning), you will
> need to remove the coverslips from some of your slides, rehydrate and then
> do your Nissl stain.   If the immunohistochemical product was oxidized DAB
> (brown), this will remain in place, and your sections will also have
> nuclear chromatin and nucleolar and cytoplasmic rRNA coloured blue, violet
> or red according to your choice of Nissl stain.
>
> Removing coverslips, extracting resinous mountant, rehydration and
> restaining is a tedious and time consuming job (days to a few
> weeks), especially if the slides are bearing thick sections (50-200um) or
> whole-mounts.
>
> As a graduate student, you should have an experienced faculty member to
> advise you.  If your boss told you to ask on the Internet, he isn't earning
> his salary.
> There are many experienced histotechnologists at the University of
> California at Davis. Histotechs love to pass on their experience, learning
> and practical advice.  You should still chase up your academic supervisor,
> who is paid from your "tuition" fees.
>
> John Kiernan
> Anatomy, UWO
> London, Canada
> = = =
> On 04/12/11, *Teresa Iglesias *<tliglesias <@t> ucdavis.edu> wrote:
>
> Hi all,
> I'm new to this so this may be a very dumb question but here goes.
> If you have already stained for IEGs (Zenk and cFos protein) in brain
> tissue and adhered coverslips with permount to the slides is it possible to
> then stain with nissl (after removing the covers, of course)?
>
> I'm having a hard time locating certain nuclei now (they were obvious
> before staining for IEGs and mounting) so I'm looking for a way to
> corroborate that I am looking in the right areas.
>
> As an alternative, I can stain new sections with nissl that have been kept
> in cryoprotectant and are free of IEG staining. IF I take this route, would
> I have to stain a replicate set of sections for ALL individuals (birds) so
> that I compare each IEG stained section to a nissl section that came from
> the same bird? In other words, I have 100 slides with 3+ sections per slide
> with IEG staining, will I need 100 slides with the same sections stained
> for nissl?
>
> Thanks for the help!!
>
> -Teresa
>
> --
> ______________________________
> Teresa Iglesias
> Graduate Group in Animal Behavior
> Department of Evolution and Ecology
> University of California-Davis
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>
>



-- 
______________________________
Teresa Iglesias
Graduate Group in Animal Behavior
Department of Evolution and Ecology

University of California-Davis
One Shields Avenue
2320 Storer Hall
Davis, CA 95616
Office: 530-754-7837
Fax: 530-752-1449
tliglesias <@t> ucdavis.edu
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