Serial Block Face Sem
Serial Block Face Sem - Related Videos
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.
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.
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.
09:02
SEM Imaging of Biological Samples
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2023
Source: Peiman Shahbeigi-Roodposhti and Sina Shahbazmohamadi, Biomedical Engineering Department, University of Connecticut, Storrs, Connecticut A scanning electron microscope (SEM) is an instrument that uses an electron beam to nondestructively image and characterize conductive materials in a vacuum. As an analogy, an electron beam is to the SEM as light is to the optical microscope. The difference is that the electron microscope yields images of much higher resolution and magnification. The...
11:41
Scanning Electron Microscopy (SEM)
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2023
Source: Laboratory of Dr. Andrew J. Steckl — University of Cincinnati A scanning electron microscope, or SEM, is a powerful microscope that uses electrons to form an image. It allows for imaging of conductive samples at magnifications that cannot be achieved using traditional microscopes. Modern light microscopes can achieve a magnification of ~1,000X, while typical SEM can reach magnifications of more than 30,000X. Because the SEM doesn’t use light to create images, the resulting pictures it...
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Each imaging cycle records the newly exposed surface after the automated ultramicrotome removes a thin layer from the resin-embedded specimen. Because successive images represent neighboring planes, computational alignment can place them in their original spatial sequence. This preserves continuity across cells, organelles, membranes, and tissue architectures, allowing researchers to follow structures through depth rather than interpreting isolated sections.
Resin embedding provides the solid block required for repeated cutting inside the scanning electron microscope. It supports the specimen as successive layers are removed and exposes a stable block face for imaging after each cut. This preparation therefore makes it possible to combine fine ultrastructural observation with volumetric reconstruction across relatively large biological tissue volumes.
Computational alignment organizes the serial images so that corresponding features occupy consistent positions across successive sections. Without this step, small shifts between images could disrupt the apparent continuity of membranes, organelles, or cells. The aligned series forms a coherent three-dimensional volume, enabling structural tracing and interpretation of how fine features are arranged within larger tissue architectures.
A typical workflow embeds the biological specimen in resin, places the block inside a scanning electron microscope, and uses an automated ultramicrotome to remove successive thin layers. After each cut, the exposed block face is imaged. The resulting serial images are then computationally aligned and reconstructed into a three-dimensional volume for structural analysis.
Researchers can choose SBF-SEM when they need to connect nanoscale ultrastructure with organization across a larger tissue volume. The method is especially relevant for tracing cells, organelles, and membranes through complex specimens. Its volumetric perspective supports investigations of neural circuits, development, disease, and cellular organization where isolated two-dimensional views may not capture structural relationships.
Serial Block Face Sem can reveal how individual cells and organelles are positioned, connected, and distributed within complex tissue architectures. Three-dimensional reconstruction also helps researchers trace membranes and cellular structures through depth, linking local ultrastructural details to broader organization. These outcomes provide a structural basis for studying biological function in neural, developmental, disease-related, and cellular contexts.