Analysis of specificity in immunoelectron microscopy

Christian Hacker, John M Lucocq

Research output: Contribution to journalArticlepeer-review

6 Citations (Scopus)

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.
Original languageEnglish
Pages (from-to)315-23
Number of pages9
JournalMethods in molecular biology (Clifton, N.J.)
Volume1117
DOIs
Publication statusPublished - 2014

Keywords

  • Animals
  • Microscopy, Immunoelectron
  • Microtomy
  • Sensitivity and Specificity
  • Staining and Labeling

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