Name: brain banking: making the most of your research specimens
Uploaded: 2009-07-24T16:02:00
Description: Watch this Scientific Journal Video about Brain Banking: Making the Most of your Research Specimens at JoVE.com

<p class="jove_content">The authors would like to thank Ikiel Ptito for his continued technical support.</p>

<p class="jove_title">Part 1: Pre-processing of tissue</p>
<ol>
<li>Tissue should be well perfused with paraformaldehyde, glutaraldehyde, or formalin. This can be achieved through standard transcardial perfusion typically used to harvest other organs. In the present study the subject was deeply sedated with ketamine hydrochloride (10 mg/kg, i.m.), euthanized with an overdose of sodium pentobarbital (25 mg/kg, i.v.) and perfused transcardially with 0.1 M PBS until completely exsanguinated. This is followed by a 4% paraformaldehyde solution in PB...

<ol>
	<li>West, M., Slomianka, L., Gundersen, H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Unbiased+stereological+estimation+of+the+total+number+of+neurons+in+the+subdivisions+of+the+rat+hippocampus+using+the+optical+fractionator.+Aat+Rec.">Unbiased stereological estimation of the total number of neurons in the subdivisions of the rat hippocampus using the optical fractionator. Aat Rec.</a> <b>231</b>, 482-497 (1991).</li><li>Burke, M. W., Zangenehpour, S., Boire, D., Ptito, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Dissecting+the+Non-human+Primate+Brain+in+Stereotaxic+Space.">Dissecting the Non-human Primate Brain in Stereotaxic Space.</a> <em style='font-style: italic'>J Vis Exp</em>. , (2009).</li></ol>

<p class="jove_content">Systematic sampling is an inexpensive method intended to maximize research material. This sampling strategy is designed to comply with the rules of unbiased stereology that requires systematic sampling throughout the region. It is critical that the order of sections for each series is maintained. In our laboratory we have successfully used this method for banking both hamster and non-human primate brain sections. So far, we have collected close to 20,000 vervet monkey (<em>Chlorocebus aethiops sabeus</em>) brain sections...

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brain banking: making the most of your research specimens

Unbiased stereology is a method for accurately and efficiently estimating the total neuron number (or other cell type) in a given area of interest<sup>1</sup>.  To achieve this goal 6-10 systematic sections should be probed covering the entire structure. Typically this involves processing 1/5 sections which leaves a significant amount of material unprocessed.  In order to maximize the material, we propose an inexpensive method for preserving fixed tissue as part of a long-term storage research plan.  As tissue is sliced and processed for the desired stain or antibody, alternate sections should be systematically placed in antigen preserve at -20&deg;C for future processing. Using 24-well plates, sections can be placed in order for future retrieval. Using this method, tissue can be stored and processed for immunohistochemistry over the course of years.

Watch this Scientific Journal Video about Brain Banking: Making the Most of your Research Specimens at JoVE.com

Brain Banking: Making the Most of your Research Specimens

<p class="jove_content">Brain banking and systematic sampling of biological material provides the basis for unbiased stereology and maximizes the potential data obtained from each specimen.</p>

Unbiased stereology is a method for accurately and efficiently estimating the total neuron number (or other cell type) in a given area of interest1.  To ...

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