AUTHOR=Petralia Ronald S. , Wang Ya-Xian TITLE=Review of Post-embedding Immunogold Methods for the Study of Neuronal Structures JOURNAL=Frontiers in Neuroanatomy VOLUME=15 YEAR=2021 URL=https://www.frontiersin.org/journals/neuroanatomy/articles/10.3389/fnana.2021.763427 DOI=10.3389/fnana.2021.763427 ISSN=1662-5129 ABSTRACT=
The post-embedding immunogold (PI) technique for immunolabeling of neuronal tissues utilizing standard thin-section transmission electron microscopy (TEM) continues to be a prime method for understanding the functional localization of key proteins in neuronal function. Its main advantages over other immunolabeling methods for thin-section TEM are (1) fairly accurate and quantifiable localization of proteins in cells; (2) double-labeling of sections using two gold particle sizes; and (3) the ability to perform multiple labeling for different proteins by using adjacent sections. Here we first review in detail a common method for PI of neuronal tissues. This method has two major parts. First, we describe the freeze-substitution embedding method: cryoprotected tissue is frozen in liquid propane