Related Experiment Video
Updated: Jul 3, 2026

11:19
Multimodal Hierarchical Imaging of Serial Sections for Finding Specific Cellular Targets within Large Volumes
Published on: March 20, 2018
Immunolabeling for scanning electron microscopy (SEM) and field emission SEM.
1Department of Biological and Biomedical Sciences, University of Durham, Durham DH1 3LE, United Kingdom.
Methods in Cell Biology
|July 12, 2008
Summary
Immuno-Scanning Electron Microscopy (Immuno-SEM) allows researchers to visualize specific proteins on cell surfaces with high resolution. This technique adapts immuno-fluorescence methods for SEM, providing context within cellular structures.
Area of Science:
- Cell Biology
- Microscopy Techniques
- Immunocytochemistry
Background:
- Scanning Electron Microscopy (SEM) offers high-resolution surface imaging.
- Many biological processes and structures are surface-dependent.
- Antibodies can localize specific components within surface structures.
Purpose of the Study:
- To adapt immuno-fluorescence techniques for SEM.
- To enable high-resolution localization of cellular components on surfaces.
- To provide context for protein localization within cellular structures.
Main Methods:
- Utilizing unconjugated primary antibodies followed by tagged secondary antibodies.
- Employing colloidal gold particles as tags for SEM detection.
- Adapting established immuno-fluorescence protocols with specific precautions.
Main Results:
- Successful adaptation of immuno-fluorescence for SEM.
- Localization of antibody-defined components within surface structures.
- Enhanced contextualization of protein location relative to cellular morphology.
Conclusions:
- Immuno-SEM is a straightforward adaptation of immuno-fluorescence.
- This method places labeling within the context of cellular structures.
- Detailed principles and practices for sample preparation, labeling, and imaging are provided.
More Related Videos
Related Concept Videos
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.
Scanning Electron Microscopy
A scanning electron microscope (SEM) is used to study the surface features of a sample by using an electron beam that scans the sample surface in a two-dimensional manner. Typically, areas between ~1 centimeter to 5 micrometers in width can be imaged. SEM can be used to image bacteria, viruses, tissues as well as larger samples like insects. Conventional SEM gives a magnification ranging from 20X to 30,000X and spatial resolution of 50 to 100 nanometers.
Fundamental Principles
Accelerated...
Fundamental Principles
Accelerated...
Preparation of Samples for Electron Microscopy
To be visualized by an electron microscope, either transmission or scanning, biological samples need to be fixed (stabilized) so the electron beam does not destroy them and dried thoroughly (desiccated/dehydrated) so the vacuum does not affect them. Fixation needs to be done as quickly as possible because the sample properties will start changing as soon as it is removed from its natural environment. For example, in a tissue sample, the oxygen levels begin decreasing, causing an altered...
Overview of Electron Microscopy
The wavelengths of visible light ultimately limit the maximum theoretical resolution of images created by light microscopes. Most light microscopes can only magnify 1000X, and a few can magnify up to 1500X. Electrons, like electromagnetic radiation, can behave like waves, but with wavelengths of 0.005 nm, they produce significantly greater resolution up to 0.05 nm as compared to 500 nm for visible light. An electron microscope (EM) can create a sharp image that is magnified up to 2,000,000X.

