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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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Updated: Aug 2, 2025

Synthesis of Near-Infrared Emitting Gold Nanoclusters for Biological Applications

Synthesis of Near-Infrared Emitting Gold Nanoclusters for Biological Applications

Published on: March 22, 2020

In situ Microinflammation Detection Using Gold Nanoclusters and a Tissue-clearing Method.

Fayrouz Naim^1,2, Rie Hasebe^1,3, Shintaro Hojyo^1,4

¹Division of Molecular Psychoneuroimmunology, Institute for Genetic Medicine, Graduate School of Medicine, Hokkaido University, Sapporo, Hokkaido, Japan.

|April 14, 2023

Summary

Researchers developed a novel 3D method to visualize microinflammation using gold nanoclusters and tissue clearing. This technique accurately maps inflammation sites in whole organs, aiding the study of inflammatory diseases like multiple sclerosis.

Keywords:

Au13 nanoclusters CD4+ T cells CUBIC Experimental autoimmune encephalomyelitis (EAE)Gateway reflex Microinflammation Myelin oligodendrocyte glycoprotein (MOG)

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Last Updated: Aug 2, 2025

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Published on: March 22, 2020

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Published on: May 21, 2015

Area of Science:

Immunology
Biomedical Engineering
Neuroscience

Background:

Microinflammation increases blood vessel permeability, mediated by inflammatory factors like IL-6 and TNF-α.
Pathogenic immune cells, such as CD4+ T cells, accumulate at specific sites, causing microinflammation and disease onset, as observed in experimental autoimmune encephalomyelitis (EAE) models of multiple sclerosis (MS).
Traditional immunohistochemistry on tissue sections is limited for identifying microinflammation sites within whole organs.

Purpose of the Study:

To develop a novel 3D visualization method for microinflammation using luminescent gold nanoclusters (AuNCs) and tissue clearing.
To enable precise localization and tracking of microinflammation development in whole organs.
To improve the understanding of inflammatory disease pathogenesis.

Main Methods:

Developed a protocol combining luminescent gold nanoclusters (AuNCs) with the CUBIC (clear, unobstructed brain/body imaging cocktails and computational analysis) tissue-clearing method.
Administered ultrasmall coordinated Au13 nanoclusters intravenously to EAE mice.
Utilized light sheet microscopy to detect leaked AuNCs from blood vessels, indicating enhanced vascular permeability at inflammation sites.

Main Results:

Successfully visualized microinflammation sites in whole spinal cords of EAE mice.
Detected leaked Au signals specifically at the dorsal vessels of the fifth lumbar cord (L5), confirming previous findings.
Demonstrated the capability to visualize complex tissue structures at the whole organ level.

Conclusions:

The developed 3D method effectively visualizes microinflammation by detecting AuNCs leaked from permeable blood vessels.
This technique allows for the precise spatial and temporal tracking of microinflammation in whole organs.
The method holds significant potential for studying various inflammatory diseases by identifying specific blood vessel sites involved in pathogenic immune cell recruitment.