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. 2016 Feb 16;10:5. doi: 10.3389/fncel.2016.00005

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Copyright © 2016 Alshammari, Alshammari and Laezza.

This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) or licensor are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.

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Scheme and examples of routinely used brain tissue preparations for immunohistochemistry. (A) Free-floating sections of 4% PFA perfusion-fixed mouse brain tissue are depicted in individual wells in a 24-well plate before mounting. (a) Representative confocal image of 4% PFA perfusion-fixed sections of the hippocampal DG immunolabeled with a rabbit anti-calbindin (CB) antibody visualized with an Alexa 488-conjugated secondary antibody (green), a mouse monoclonal anti-calretinin (CR) antibody visualized with an Alexa 568-conjugated secondary antibody (red) and Topro-3 nuclei staining (blue). (B) Fresh-frozen mouse brain slices directly adhered to positively charged glass slides. (b) Representative confocal image of a fresh-frozen section of the hippocampal DG, immunolabeled with a mouse monoclonal anti-FGF14 antibody visualized with an Alexa 568-conjugated secondary antibody. PFA, paraformaldehyde; DG, dentate gyrus; ML, molecular layer; GCL, granule cell layer; SGZ, sub-granular zone. Scale bars represent 20 μm.