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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.
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Analysis of Brain Mitochondria Using Serial Block-Face Scanning Electron Microscopy
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Analysis of Brain Mitochondria Using Serial Block-Face Scanning Electron Microscopy

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Serial block face-scanning electron microscopy for volume electron microscopy.

Saskia Lippens1, Anna Kremer1, Peter Borghgraef1

  • 1VIB BioImaging Core, VIB, Ghent, Belgium; VIB Inflammation Research Center, VIB, Ghent, Belgium; Department of Biomedical Molecular Biology, Ghent University, Ghent, Belgium.

Methods in Cell Biology
|July 22, 2019
PubMed
Summary
This summary is machine-generated.

Serial Block Face Scanning Electron Microscopy (SBF-SEM) offers nanometer-resolution 3D imaging across various life science fields. This guide details the SBF-SEM workflow, highlighting adaptable steps for diverse sample types.

Keywords:
Focal charge compensatorSample preparationSerial block-face scanning electron microscopyThree dimensional electron microscopyVolume electron microscopy

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Area of Science:

  • Life Science Research
  • Microscopy Techniques
  • Nanotechnology

Background:

  • Serial Block Face Scanning Electron Microscopy (SBF-SEM) is increasingly used for 3D imaging in life sciences.
  • Originally developed for neuroscience, its application has broadened to other research areas.
  • SBF-SEM enables the capture of 3D volumes from monolayers to multiple tissue layers with nanometer resolution.

Purpose of the Study:

  • To describe the Serial Block Face Scanning Electron Microscopy (SBF-SEM) workflow.
  • To identify and explain adjustable steps within the SBF-SEM sample preparation process.
  • To facilitate the adaptation of SBF-SEM for various sample types in life science research.

Main Methods:

  • Detailed description of the Serial Block Face Scanning Electron Microscopy (SBF-SEM) workflow.
  • Identification of critical steps in sample preparation amenable to modification.
  • Methodology focused on optimizing SBF-SEM for diverse biological specimens.

Main Results:

  • A comprehensive overview of the SBF-SEM process is provided.
  • Specific steps within the workflow that can be adjusted for different samples are highlighted.
  • The study offers insights into tailoring SBF-SEM protocols for broader applicability.

Conclusions:

  • The SBF-SEM technique is a powerful tool for high-resolution 3D imaging in life sciences.
  • Understanding and adapting specific workflow steps is crucial for successful SBF-SEM implementation across different research domains.
  • This work provides a valuable resource for researchers seeking to optimize SBF-SEM for their unique sample requirements.