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Pre-embedding immunoelectron microscopy of chemically fixed mammalian tissue culture cells
Haruo Hagiwara1, Takeo Aoki, Takeshi Suzuki
1Department of Occupational Therapy, School of Health Sciences, Faculty of Medicine, Gunma University, Gunma, Japan.
Insights
This study details using Nanogold particles for pre-embedding immunoelectron microscopy in cultured cells. Silver enhancement allows control over gold particle size for protein localization.
Area of Science:
- Cell Biology
- Microscopy Techniques
- Biochemistry
Background:
- Immunoelectron microscopy is crucial for protein localization in cells and tissues.
- Nanogold particles (1.4 nm) conjugated with Fab' fragments enable intracellular penetration.
- Silver enhancement is necessary to visualize gold labels and control granule size.
Purpose of the Study:
- To describe the application of Nanogold for pre-embedding immunoelectron microscopy.
- To demonstrate protein localization in paraformaldehyde-fixed cultured cells using Nanogold.
Main Methods:
- Pre-embedding immunoelectron microscopy technique.
- Utilizing Nanogold particles conjugated with Fab' fragments.
- Paraformaldehyde fixation of cultured cells.
- Silver enhancement for gold particle visualization and size control.
Main Results:
- Nanogold particles successfully penetrated paraformaldehyde-fixed cultured cells.
- Silver enhancement allowed for controlled visualization of gold labels.
- Protein localization was achieved using this method.
Conclusions:
- Nanogold is effective for pre-embedding immunoelectron microscopy in cultured cells.
- Silver enhancement offers tunable contrast for gold labels.
- This technique provides a reliable method for intracellular protein detection.
Abstract:
Immunoelectron microscopy is one of the best methods for detecting and localizing protein molecules in cells and tissues. Gold particles of 1.4 nm in diameter (Nanogold) conjugated with Fab' fragments easily penetrate into the cell interior and are used for pre-embedding immunoelectron microscopy. To obtain a contrast for the gold label, silver enhancement of the gold particles is essential. By changing the intensity of the silver enhancement, the size of the granules can be controlled. In this chapter, we described the use of Nanogold for pre-embedding immunoelectron microscopy of paraformaldehyde-fixed cultured cells.
