Serial Block-face Microscopy
Serial Block-face Microscopy - Related Videos
07:47
Analysis of Brain Mitochondria Using Serial Block-Face Scanning Electron Microscopy
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Cited by 32 •
2016
Mitochondrial visualization and analysis from mammalian brain tissue is a challenging task. Here, we describe how three dimensional (3D) reconstruction analysis from the serial block-face scanning electron microscopy (SBFSEM) can be used to gain insights on the morphological and volumetric analysis of this critical energy generating organelle.
11:16
Serial Block-Face Scanning Electron Microscopy (SBEM) for the Study of Dendritic Spines
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Cited by 7 •
2021
Serial Block-Face Scanning Electron Microscopy (SBEM) is applied to image and analyze dendritic spines in the murine hippocampus.
09:21
Serial Block-Face Scanning Electron Microscopy (SBF-SEM) of Biological Tissue Samples
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Cited by 32 •
2021
This protocol outlines a routine method for using serial block-face scanning electron microscopy (SBF-SEM), a powerful 3D imaging technique. Successful application of SBF-SEM hinges on proper fixation and tissue staining techniques, as well as careful consideration of imaging settings. This protocol contains practical considerations for the entirety of this process.
03:25
Imaging of Neuronal Mitochondria Using a Serial Block-Face Scanning Electron Microscope
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2025
Source: Mukherjee, K., et. al. Analysis of Brain Mitochondria Using Serial Block-Face Scanning Electron Microscopy. J. Vis. Exp. (2016)This video demonstrates the preparation, imaging, and analysis of mouse brain tissue using a serial block-face scanning electron microscope (SBFSEM) to visualize mitochondrial ultrastructure, enabling precise mapping of their morphology and spatial distribution in axonal and dendritic neuronal compartments.
09:09
Targeted Studies Using Serial Block Face and Focused Ion Beam Scan Electron Microscopy
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Cited by 15 •
2019
Here, we present a protocol for efficiently combining serial block face and focused ion beam scanning electron microscopy to target an area of interest. This allows for efficient searching, in three dimensions, and locating rare events in a large field of view.
FAQs
The method builds three-dimensional information from a sequence of surface views rather than from a single image. After the electron beam records one exposed face, the integrated ultramicrotome removes a thin layer and reveals the next face. Repeating this cycle preserves the order of structures through depth, allowing the resulting images to represent a continuous tissue volume.
The integrated ultramicrotome controls the transition between successive imaging surfaces. It removes a thin layer after each electron-microscope scan, exposing new ultrastructure for the next image. This coordinated cutting and imaging sequence links observations from neighboring levels of the specimen, which is essential for following neurites, synapses, organelles, and other structures through the reconstructed volume.
Computational alignment places successive images into their correct spatial relationships before reconstruction. Without this step, structures appearing in adjacent tissue layers could be misregistered, making their continuity difficult to interpret. Once aligned, the dataset supports three-dimensional visualization and quantitative analysis of cellular architecture, including the paths and relationships of fine neural structures.
A typical workflow begins with an embedded biological specimen positioned for electron microscopy. The beam scans the exposed tissue surface, the integrated ultramicrotome removes a thin layer, and the newly exposed surface is imaged again. After many cycles, computational alignment organizes the image series into a three-dimensional representation suitable for examining ultrastructure across the sampled volume.
Researchers can use Serial Block-face Microscopy when they need to trace fine neural structures across a volume rather than inspect isolated profiles. Its serial images support mapping neurons, neurites, and synapses, making it useful for connectomics and circuit reconstruction. The reconstructed architecture can reveal how cellular components are arranged and connected within brain tissue.
The datasets provide detailed structural measurements that can be compared with questions about neural function and disease. By mapping cellular architecture across brain tissue, investigators can examine how neurons, neurites, synapses, and organelles are organized in relevant regions. This creates a structural basis for quantitative studies linking microscopic anatomy with functional or disease-related changes.