A freeze substitution fixation-based gold enlarging technique for EM studies of endocytosed Nanogold-labeled molecules.

ABSTRACT: We have developed methods to locate individual ligands that can be used for electron microscopy studies of dynamic events during endocytosis and subsequent intracellular trafficking. The methods are based on enlargement of 1.4 nm Nanogold attached to an endocytosed ligand. Nanogold, a small label that does not induce misdirection of ligand-receptor complexes, is ideal for labeling ligands endocytosed by live cells, but is too small to be routinely located in cells by electron microscopy. Traditional pre-embedding enhancement protocols to enlarge Nanogold are not compatible with high pressure freezing/freeze substitution fixation (HPF/FSF), the most accurate method to preserve ultrastructure and dynamic events during trafficking. We have developed an improved enhancement procedure for chemi

SUBMITTER: He W 

PROVIDER: S-EPMC2076746 | biostudies-literature | 2007 Oct

REPOSITORIES: biostudies-literature