[Histonet] nissl after permount?

[John Kiernan]

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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