I am trying to collect the hippocampus, prefrontal cortex and amygdala from adolescent rat brains (unfixed). Is there a way I could validate the accuracy of the dissection to rule out contamination from adjacent brain regions? For instance, I have been searching for molecular markers of the hippocampus that are absent in adjacent regions, and vice versa.
Thank you
You might want to try taking a slice however many microns thick freeze it down onto a microscope slide then punching out or using a blade to cut out your region of interest. You can then take the slide to a microscope and snap a quick picture to ensure you're not getting too much surrounding tissue. This is how I did amygdala punches in adult rats for years. I'm not sure you can really get clean molecular markers for hippocampus vs PFC vs amygdala.
Its very difficult to get molecular markers for particular brain region (receptors distributed through out brain). I feel in initial stage you can cross check your sectioning with binding study. Like in hippocampus, amygdala and PFC receptor expression will be different and specific compound binding also varies. Thus you can cross check your isolated region is specific or not. After having adequate hands on training no need to worry about marker or binding. Practice and concentration is the only way in these kind of situation.
Regards
Ramanatha
Take the slice from which you dissect your regions of interest, fix the remaining tissue and slice using a microtome/cryostat. Then stain the section using e.g. cresyl violet and validate based on anatomy
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