A Scalable Histological Method to Embed and Section Multiple Brains Simultaneously

Author:

Nwafor Divine C.1,Benkovic Stanley A.2,Clary Briana L.23ORCID,Brichacek Allison L.4,Lambert H. Wayne5ORCID,Zdilla Matthew J.5ORCID,Brown Candice M.234ORCID

Affiliation:

1. Department of Neurosurgery, University of Virginia, Charlottesville, VA 22903, USA

2. Department of Neuroscience, School of Medicine, West Virginia University, Morgantown, WV 26506, USA

3. Rockefeller Neuroscience Institute, West Virginia University, Morgantown, WV 26506, USA

4. Department of Microbiology, Immunology, and Cell Biology, School of Medicine, West Virginia University, Morgantown, WV 26506, USA

5. Department of Pathology, Anatomy and Laboratory Medicine, School of Medicine, West Virginia University, Morgantown, WV 26506, USA

Abstract

The preparation and processing of rodent brains for evaluation by immunohistochemistry is time-consuming. A large number of mouse brains are routinely used in experiments in neuroscience laboratories to evaluate several models of human diseases. Thus, methods are needed to reduce the time associated with processing brains for histology. A scalable method was developed to embed, section, and stain multiple mouse brains using supplies found in any common histology laboratory. Section collection schemes can be scaled to provide identical bregma locations between adjacent sections for immunohistochemistry, facilitating comprehensive, high-quality immunohistochemistry. As a result, sectioning and staining times are considerably reduced as sections from multiple blocks are stained simultaneously. This method improves on previous procedures and allows multiple embedding and subsequent immunostaining of brains easily with a dramatically reduced time requirement. Furthermore, we expand this method for use in numerous mouse tissues, rat brain tissue, and post-mortem human brain and arterial tissues. In summary, this procedure allows the processing of many rodent or human tissues from perfusion through microscopy in 10 days or less.

Funder

NIH

Publisher

MDPI AG

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