Related Experiment Video

Updated: May 24, 2026

Nanogold Labeling of the Yeast Endosomal System for Ultrastructural Analyses
09:49

Nanogold Labeling of the Yeast Endosomal System for Ultrastructural Analyses

Published on: July 14, 2014

Immunogold probes for light microscopy

J E Beesley1

  • 1Wellcome Research Laboratories, Beckenham, UK.

Methods in Molecular Biology (Clifton, N.J.)
|February 22, 2012
PubMed

Insights

Immunocytochemistry techniques use microscopic markers to locate antigens. Advances include immunofluorescence, enzyme-based methods like peroxidase and alkaline phosphatase, and newer colloidal gold probes for light microscopy.

Area of Science:

  • Immunocytochemistry
  • Microscopy
  • Biochemical Assays

Background:

  • Immunocytochemistry enables antigen localization within cells and tissues.
  • Early methods relied on immunofluorescence, pioneered by Coons et al.
  • Subsequent developments introduced enzyme-based markers for enhanced sensitivity.

Purpose of the Study:

  • To review the evolution of immunocytochemical techniques.
  • To highlight the development of various microscopic markers.
  • To discuss the application of new markers at the light microscope level.

Main Methods:

  • Review of historical and contemporary immunocytochemical methodologies.
  • Description of immunofluorescence, immunoenzyme, and colloidal gold probe techniques.
  • Discussion of antigen localization strategies using different markers.

Main Results:

  • Immunofluorescence was the foundational technique for antigen localization.
  • Enzyme-based methods, such as peroxidase and alkaline phosphatase, offer sensitive detection.
  • Colloidal gold probes are emerging as effective markers for light microscopy.

Conclusions:

  • Immunocytochemical techniques have evolved significantly with diverse marker systems.
  • Enzyme and colloidal gold markers provide valuable alternatives to immunofluorescence.
  • These advancements enhance the capability of antigen localization in biological research.

More Related Videos

High-Resolution Quantitative Immunogold Analysis of Membrane Receptors at Retinal Ribbon Synapses09:24

High-Resolution Quantitative Immunogold Analysis of Membrane Receptors at Retinal Ribbon Synapses

Published on: February 18, 2016

Correlative Light- and Electron Microscopy Using Quantum Dot Nanoparticles11:16

Correlative Light- and Electron Microscopy Using Quantum Dot Nanoparticles

Published on: August 7, 2016

Related Concept Videos

Immunogold Electron Microscopy01:20

Immunogold Electron Microscopy

Immunoelectron microscopy utilizes immunogold labeling of endogenous proteins with specific antibodies to detect and localize these proteins in cells and tissues. The procedure provides insights into the distribution and quantification of protein under different stimulation conditions offering clues about their functions. Conjugating highly electron-dense gold particles with primary or secondary antibodies allow antigen detection on and within cells, with high resolution and specificity.
Immunofluorescence Microscopy01:12

Immunofluorescence Microscopy

A fluorescence microscope uses fluorescent chromophores called fluorochromes, which can absorb energy from a light source and then emit this energy as visible light. Fluorochromes include naturally fluorescent substances (such as chlorophylls) and fluorescent stains that are added to the specimen to create contrast. Dyes such as Texas red and FITC are examples of fluorochromes. Other examples include the nucleic acid dyes 4’,6’-diamidino-2-phenylindole (DAPI), and acridine orange.
The...
Labeling DNA Probes03:31

Labeling DNA Probes

DNA probes are fragments of DNA labeled with a reporter tag to enable their detection or purification. The resulting labeled DNA probes can then hybridize to target nucleic acid sequences through complementary base-pairing, and may be used to recover or identify these regions.
Radioisotopes, fluorophores, or small molecule binding partners like biotin or digoxigenin, are the most widely used reporter tags for labeling DNA probes. These labels can be attached to the probe DNA molecule via...
Overview of Microscopy Techniques01:22

Overview of Microscopy Techniques

The early pioneers of microscopy opened a window into the invisible world of microorganisms. In 1830, Joseph Jackson Lister created an essentially modern light microscope. The 20th century saw the development of microscopes that leveraged nonvisible light, such as fluorescence microscopy that uses an ultraviolet light source and electron microscopy that uses short-wavelength electron beams. These advances significantly improved magnification, image resolution, and contrast. By comparison, the...