Related Experiment Video
Updated: Jan 20, 2026
09:09
Targeted Studies Using Serial Block Face and Focused Ion Beam Scan Electron Microscopy
Published on: August 10, 2019
9.7K
Targeted Studies Using Serial Block Face and Focused Ion Beam Scan Electron Microscopy.
Christopher J Guérin1, Anna Kremer1, Peter Borghgraef1
1VIB Bio Imaging Core; VIB Inflammation Research Center; Department of Molecular Biomedical Research, UGent.
Journal of Visualized Experiments : Jove
|August 27, 2019
Summary
This protocol enables high-resolution 3D electron microscopy (EM) imaging by combining serial block face scanning EM (SBF-SEM) and focused ion beam SEM (FIB-SEM). Microwave-assisted processing significantly reduces sample preparation time from days to hours.
Area of Science:
- Cell Biology
- Microscopy Techniques
- Biotechnology
Background:
- Electron microscopy (EM) traditionally offers 2D imaging.
- Volume EM techniques like SBF-SEM and FIB-SEM enable 3D imaging.
- Current 3D EM methods have limitations in field of view or Z-axis resolution.
Purpose of the Study:
- To present a protocol for high-resolution 3D electron microscopy imaging.
- To combine SBF-SEM and FIB-SEM for comprehensive 3D analysis.
- To reduce sample preparation time for volume EM.
Main Methods:
- Serial block face scanning electron microscopy (SBF-SEM) for large fields of view.
- Focused ion beam scanning electron microscopy (FIB-SEM) for high isotropic resolution.
- Microwave-assisted tissue processing for enhanced reagent penetration and reduced preparation time.
Main Results:
- Successful integration of SBF-SEM and FIB-SEM workflows.
- Achieved high isotropic voxel resolution (≤5 nm) in targeted volumes.
- Reduced sample processing time from days to hours.
Conclusions:
- The combined SBF-SEM and FIB-SEM approach allows for efficient 3D imaging of large samples with high resolution.
- Microwave-assisted processing streamlines sample preparation for volume EM.
- This protocol enhances the capability of electron microscopy for detailed cellular and tissue analysis.
Related Concept Videos
01:07Scanning Electron Microscopy
5.3K
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...
5.3K
01:25Overview of Electron Microscopy
13.1K
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.
13.1K
01:15Transmission Electron Microscopy
6.9K
In 1931, physicist Ernst Ruska—building on the idea that magnetic fields can direct an electron beam just as lenses can direct a beam of light in an optical microscope—developed the first prototype of the electron microscope. This development led to the development of the field of electron microscopy. In the transmission electron microscope (TEM), electrons are produced by a hot tungsten element and accelerated by a potential difference in an electron gun, which gives them up to 400...
6.9K
01:20Immunogold Electron Microscopy
5.3K
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.
5.3K
01:28Cryo-electron Microscopy
4.2K
Conventional electron microscopy (EM) involves dehydration, fixation, and staining of biological samples, which distorts the native state of biological molecules and results in several artifacts. Also, the high-energy electron beam damages the sample and makes it difficult to obtain high-resolution images. These issues can be addressed using cryo-EM, which uses frozen samples and gentler electron beams. The technique was developed by Jacques Dubochet, Joachim Frank, and Richard Henderson, for...
4.2K
03:27Ions and Ionic Charges
78.6K
In ordinary chemical reactions, the nucleus — which contains the protons and neutrons of each atom and thus identifies the element — remains unchanged. Electrons, however, can be added to atoms by transfer from other atoms, lost by transfer to other atoms, or shared with other atoms. The transfer and sharing of electrons among atoms govern the chemistry of the elements. During the formation of some compounds, atoms gain or lose electrons to form electrically charged particles called...
78.6K