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Immunogold Electron Microscopy

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.

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Related Experiment Video

Updated: May 30, 2026

Rapid Nanoprobe Signal Enhancement by In Situ Gold Nanoparticle Synthesis

Rapid Nanoprobe Signal Enhancement by In Situ Gold Nanoparticle Synthesis

Published on: March 7, 2018

Enhanced solid-phase immunoassay using gold nanoshells: effect of nanoparticle optical properties.

Boris Khlebtsov¹, Nikolai Khlebtsov

¹Institute of Biochemistry and Physiology of Plants and Microorganisms, Russian Academy of Sciences, 13 Prospekt Entuziastov, Saratov 410049, Russia.

|August 12, 2011

Summary

This study introduces a theoretical model for nanoparticle-labeled immunoassays, demonstrating significantly enhanced detection sensitivity using silica/gold nanoshells (GNs) compared to traditional gold nanoparticles for IgG detection.

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Area of Science:

Nanotechnology
Biochemistry
Analytical Chemistry

Background:

Plasmon-resonant nanoparticle-labeled immunoassays offer a cost-effective method for detecting molecules.
The underlying optical mechanisms driving their efficiency remain largely unaddressed.
This research addresses the theoretical and experimental aspects of nanoparticle-based immunoassay sensitivity.

Purpose of the Study:

To provide the first theoretical description of nanoparticle-labeled dot immunoassays.
To experimentally verify the theoretical model using colloidal gold nanospheres and silica/gold nanoshells (GNs).
To investigate the impact of nanoparticle properties on immunoassay detection limits and sensitivity.

Main Methods:

Development of a theoretical framework for nanoparticle-labeled dot immunoassays.
Fabrication and characterization of 15 nm colloidal gold nanospheres and silica/gold nanoshells (100/15, 140/15, 180/15 nm) using microscopy and spectroscopy.
Experimental verification using a model system of rabbit IgG and sheep antirabbit antibodies.

Main Results:

The study presents a theoretical model explaining the optical mechanisms behind nanoparticle immunoassay efficiency.
Silica/gold nanoshells significantly enhanced detection sensitivity compared to 15 nm gold nanoparticles, reducing the detection limit for IgG to as low as 0.25 ng (180/15 nm GNs).
Theoretical predictions accurately explained the observed relationships between particle parameters and assay performance metrics like detection limit and probe-load saturation.

Conclusions:

The developed theory provides a fundamental understanding of plasmon-resonant nanoparticle-labeled immunoassays.
Silica/gold nanoshells represent a superior labeling agent for enhancing immunoassay sensitivity.
The findings enable the optimization of nanoparticle design for improved diagnostic applications.