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Focused Ion Beam Scanning Electron Microscopy
High-Resolution, Isotropic Volume Data for Accurate 3D Reconstructions
Schematic Representation of a Typical Workflow
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1
A trench is milled into a resin-embedded sample with a focused ion beam until the structure of interest becomes visible.
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2
The newly exposed sample surface of the structure of interest is imaged. This milling and imaging process is repeated until the structure is completely imaged.
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3
The acquired EM images are processed and digitally aligned into a 3D data set. Cell compartments can be identified and segmented.
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4
The segmented 3D data set can be visualized, investigated, and statistically analyzed.
Application Examples
High-Resolution, Isotropic Visualization of Cellular Ultrastructure in 3D
3D Imaging of HeLa Cells
Automated 3D Serial Imaging with ZEISS FIB-SEM Technology
The focused ion beam was utilized to sequentially remove 8 nm thick layers of the specimen while the exposed block-face is scanned with a scanning electron microscope, thus obtaining a high-resolution 3D volume image. Automated segmentation and visualization of cellular components was done using an arivis Cloud-trained deep learning model in arivis Pro so that the different cellular components could be visualized and quantified.
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Image courtesy of Dr Louise Hughes, Oxford Brookes University, UK
Characterizing the Golgi Apparatus
To Better Understand Its Role in Protein Modification and Transport
This image shows a 3D reconstruction of algal Golgi body from a FIB-SEM data set. The data set distinguishes between the cis and trans faces of the Golgi (yellow/red: cis-golgi, purple/blue: trans-golgi). Segmentation of the cellular components from the high-resolution data sets acquired using ZEISS Crossbeam FIB-SEM technology ensures that internal components can be accurately characterized and quantified.