We've found that the most useful method for fluorescently tagging neurons is to use IHC with a primary against neuronal nuclear protein (NeuN). The NeuN can be processed for fluorescence or light-field and makes a fantastic counterstain, allowing the sections to be matched easily with atlas plates. If you're interested in delineating the shell and core subregions of the accumbens (and you probably should be), you might want to try processing the tissue for calcium-binding protein, which is differentially distributed in these compartments.
http://www.ncbi.nlm.nih.gov/pubmed/7526940
Your best bet to locate them is to stain with thionin or cresyl violet or something of this nature. Why do you want do do immunofluorescence, rather than a more common stain?
If you already have tissue labeled, what did you use for your immunofluorescence work?
Dear Greg, thanks for your input. I wish to look at several types of proteins expressed in those regions.
It's difficult to distinguish clearly between the two structures using immunofluorescence but as mentioned by Greg, using staining methods could help greatly. You use also enzymatic methods as acetylcholinesterase staining. Please see these references:
- http://www.ncbi.nlm.nih.gov/pubmed/15223749
-http://cshprotocols.cshlp.org/content/2010/8/pdb.prot4806.short
Either immunofluorescence or any other conventional staining, you have to look for different anatomical landmarks that are available in plenty in literature to identify the area of your interest...simultaneously use rat/mouse brain atlas .. otherwise I don't think there is a specific immunological markers for NA or vmPFC..
Hello
to localise the Nucleus Accumbens you can use/or do the superposition of your slides with an atlas of rat brain (Paxinos and Watson), by smatching diffetentes slides (virtualy) with a level of the nucleus Accumbens (surronding the anterior commisur). you can do the same for vmPFC.
We've found that the most useful method for fluorescently tagging neurons is to use IHC with a primary against neuronal nuclear protein (NeuN). The NeuN can be processed for fluorescence or light-field and makes a fantastic counterstain, allowing the sections to be matched easily with atlas plates. If you're interested in delineating the shell and core subregions of the accumbens (and you probably should be), you might want to try processing the tissue for calcium-binding protein, which is differentially distributed in these compartments.
http://www.ncbi.nlm.nih.gov/pubmed/7526940
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