Thank you for the excellent information. -Teresa On Mon, Dec 5, 2011 at 1:03 AM, John Kiernan <jkier...@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 *<tligles...@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 > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > -- ______________________________ 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 tligles...@ucdavis.edu ______________________________ _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet