Analysis of specificity in immunoelectron microscopy

Abstract

Immunoelectron microscopy (immuno-EM) using gold labeling on sections is a powerful technique for mapping the distribution of proteins, lipids, carbohydrates, and nucleic acids in intact biological systems. The gold particles comprise a useful and readily quantifiable digital readout. Simply applying a labeling reagent (antibody or other affinity probe) to an ultrathin section yields a pattern of gold signal over the biological structures displayed in the section. This initial (raw) distribution of gold signal contains both specific and nonspecific labeling. Here we describe a method for removing nonspecific labeling to leave the target-specific signal. This specific labeling distribution better reflects the "real" distribution of the cell component of interest.