Focused Ion Beam Scanning Electron Microscopy
High-Resolution, Isotropic Volume Data for Accurate 3D Reconstructions
Schematic Representation of a Typical Workflow
A trench is milled into a resin-embedded sample with a focused ion beam until the structure of interest becomes visible.
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.
The acquired EM images are processed and digitally aligned into a 3D data set. Cell compartments can be identified and segmented.
The segmented 3D data set can be visualized, investigated, and statistically analyzed.
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 APEER-trained deep learning model in Vision4D so that the different cellular components could be visualized and quantified.
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.