Processing Neonatal Mouse Brains for Immunohistochemical Analysis: As Required for Including Spindle Orientation Analysis and Analysis of DNA Damage and Apoptosis

Gabriela De la Cruz; Nana Nikolaishvili-Feinberg; Scott E Williams; Timothy R Gershon

doi:10.1007/978-1-0716-2752-5_5

Processing Neonatal Mouse Brains for Immunohistochemical Analysis: As Required for Including Spindle Orientation Analysis and Analysis of DNA Damage and Apoptosis

Methods Mol Biol. 2023:2583:49-54. doi: 10.1007/978-1-0716-2752-5_5.

Authors

Gabriela De la Cruz^{1

2}, Nana Nikolaishvili-Feinberg^{1

2}, Scott E Williams^{1

3

4}, Timothy R Gershon^{5

6

7}

Affiliations

¹ Department of Pathology & Laboratory Medicine, The University of North Carolina at Chapel Hill, Chapel Hill, NC, USA.
² Translational Pathology Laboratory, The University of North Carolina at Chapel Hill, Chapel Hill, NC, USA.
³ Department of Biology, The University of North Carolina at Chapel Hill, Chapel Hill, NC, USA.
⁴ Lineberger Comprehensive Cancer Center, The University of North Carolina at Chapel Hill, Chapel Hill, NC, USA.
⁵ Lineberger Comprehensive Cancer Center, The University of North Carolina at Chapel Hill, Chapel Hill, NC, USA. gershont@neurology.unc.edu.
⁶ Department of Neurology, The University of North Carolina at Chapel Hill, Chapel Hill, NC, USA. gershont@neurology.unc.edu.
⁷ UNC Neuroscience Center, The University of North Carolina at Chapel Hill, Chapel Hill, NC, USA. gershont@neurology.unc.edu.

PMID: 36418724
DOI: 10.1007/978-1-0716-2752-5_5

Abstract

Analyzing sections of neonatal mouse brain using immunohistochemistry can inform microcephaly pathogenesis, but obtaining and staining high-quality sections can be challenging. The neonatal brain shows less structural integrity than the adult brain. As a result, embedding technique must be optimized to allow sections without cracks or other anatomic disruptions. Moreover, paraffin embedding, which maximized tissue preservation, can reduce antigenicity of proteins in the embedded tissues. We describe an optimized embedding technique and antigen recovery technique that allows successful sectioning and immunohistochemical staining.

Keywords: Cerebellum; Granule cells; Hippocampus; Microcephaly; Neural progenitor; Postnatal neurogenesis; Subventricular zone.

MeSH terms

Animals
Animals, Newborn
Apoptosis
Brain*
DNA Damage*
Mice
Paraffin Embedding