Glyoxal Fixation Is Optimal for Immunostaining of Brain Vessels, Pericytes and Blood-Brain Barrier Proteins

Authors

Sithara Thomas
Jayanarayanan Sadanandan
Spiros L Blackburn
Devin W McBride
Ari Dienel
Sungha Hong
Hussein A Zeineddine
Peeyush Kumar Thankamani

Publication Date

7-14-2022

Journal

International Journal of Molecular Sciences

Abstract

Brain vascular staining is very important for understanding cerebrovascular pathologies. 4% paraformaldehyde is considered the gold standard fixation technique for immunohistochemistry and it revolutionized the examination of proteins in fixed tissues. However, this fixation technique produces inconsistent immunohistochemical staining results due to antigen masking. Here, we test a new fixation protocol using 3% glyoxal and demonstrate that this method improves the staining of the brain vasculature, pericytes, and tight junction proteins compared to 4% paraformaldehyde. Use of this new fixation technique will provide more detailed information about vascular protein expressions, their distributions, and colocalizations with other proteins at the molecular level in the brain vasculature.

Keywords

glyoxal, paraformaldehyde, vascular staining, blood-brain barrier, tight junction, pericytes