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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.

Immunocytochemistry and Immunohistochemistry

Immunocytochemistry and Immunohistochemistry

Immunocytochemistry (ICC) and immunohistochemistry (IHC) are techniques that use antibodies to check for specific proteins or antigens in a sample. The technique was first published by Albert Coons in 1941 to detect the presence of pneumococcal antigen in tissue sections from mice infected with Pneumococcus. Immunocytochemistry helps localization of proteins or antigens in individual cells like blood cells, stem cells, etc., while immunohistochemistry does the same for tissue samples.
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Double-Staining Method to Detect Pectin in Plant-Fungus Interaction

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Immunostaining of Ectocarpus cells.

Susana M Coelho¹, Delphine Scornet, Sylvie Rousvoal

¹UPMC Université Paris 06, The Marine Plants and Biomolecules Laboratory, UMR 7139, Station Biologique de Roscoff, 29682 Roscoff Cedex, France.

Cold Spring Harbor Protocols

|March 3, 2012

Summary

This summary is machine-generated.

This study presents an optimized immunostaining protocol for Ectocarpus, a model organism ideal for high-resolution imaging. The method facilitates antibody detection in transparent cells, aiding cellular and molecular research.

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Area of Science:

Marine Biology
Cell Biology
Microscopy

Background:

Ectocarpus exhibits small, transparent cells and uniseriate filaments.
These characteristics are advantageous for high-resolution imaging techniques.
The organism can be cultured directly on microscope slides.

Purpose of the Study:

To describe an optimized immunostaining protocol for Ectocarpus.
To enable the detection of specific proteins, such as tubulin, using antibodies.
To facilitate high-resolution imaging of Ectocarpus cells.

Main Methods:

Immunostaining protocol optimized for Ectocarpus.
Incubation steps performed on a platform shaker at room temperature.
Recommendations for using high-quality microscope slides for confocal laser scanning microscopy.

Main Results:

The protocol is effective for detecting tubulin.
The protocol is adaptable for use with various antibodies.
Ectocarpus's cellular and growth properties are suitable for advanced imaging.

Conclusions:

The described immunostaining protocol is a valuable tool for Ectocarpus research.
This method supports high-resolution in vivo and fixed imaging.
The protocol enhances the utility of Ectocarpus as a model organism in cell biology.