An improved approach to align and embed multiple brain samples in a gelatin-based matrix for simultaneous histological processing

J Neurosci Methods. 2016 Mar 1:261:155-60. doi: 10.1016/j.jneumeth.2015.12.008. Epub 2015 Dec 29.

Abstract

Background: Preparation and processing of free-floating histological sections involve a series of steps. The amount of labor, particularly sectioning and mounting, quickly multiplies as the number of samples increases. Embedding tissue samples in a flexible matrix allows simultaneous handling of multiple samples and preserves the integrity of the tissue during histological processing. However, aligning multiple asymmetrical samples, for example small-animal brains, in a particular orientation requires skillful arrangement and securing of the samples by pinning onto a solid surface. Consequently, costly technical services offered by contract research organizations are often sought.

New method: An improved approach to align and embed multiple whole or half rodent brain samples into a gelatin-based matrix is described. Using a template specifically designed to form arrayed mouse brain-shaped cavities, a "receiving matrix" is prepared. Inserting brain samples directly into the cavities allows the samples to be effortlessly positioned into a uniform orientation and embedded in a block of matrix.

Results: Multiple mouse brains were arrayed in a uniform orientation in a gelatin matrix block with ease using the receiving matrix. The gelatin-embedded brains were simultaneously sectioned and stained, and effortlessly mounted onto glass slides.

Comparison with existing methods: The improved approach allowed multiple whole or half mouse brains to be easily arrayed without pinning the samples onto a solid surface and prevented damages or shifting of the samples during embedding.

Conclusions: The new approach to array multiple brain samples provides a simple way to prepare gelatin-embedded whole or half brain arrays of commercial quality.

Keywords: Brain array; Embedding; Frozen-section; Gelatin; Immunohistochemistry; Matrix; Tissue preparation.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Brain / cytology*
  • Brain / metabolism
  • Calcium-Binding Proteins / metabolism
  • Frozen Sections / methods
  • Gelatin
  • Immunohistochemistry / methods
  • Mice, Inbred C57BL
  • Microfilament Proteins / metabolism
  • Microglia / cytology
  • Microglia / metabolism
  • Photomicrography
  • Sucrose
  • Tissue Embedding / methods*

Substances

  • Aif1 protein, mouse
  • Calcium-Binding Proteins
  • Microfilament Proteins
  • Sucrose
  • Gelatin