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Multimodal Hierarchical Imaging of Serial Sections for Finding Specific Cellular Targets within Large Volumes

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Published on: March 20, 2018

Enhanced FIB-SEM systems for large-volume 3D imaging.

C Shan Xu¹, Kenneth J Hayworth¹, Zhiyuan Lu^1,2

¹Janelia Research Campus, Howard Hughes Medical Institute, Ashburn, United States.

|May 14, 2017

Summary

Focused Ion Beam Scanning Electron Microscopy (FIB-SEM) now offers faster imaging and improved stability for large-volume 3D reconstruction. This breakthrough enables advanced connectomics and cell biology research with high-resolution neural tissue imaging.

Keywords:

3D cellular structure FIB cell biology connectomics electron microscopy imaging neuroscience

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Area of Science:

Neuroscience
Cell Biology
Microscopy

Background:

Focused Ion Beam Scanning Electron Microscopy (FIB-SEM) offers high z-axis resolution for 3D imaging.
Current limitations include slow imaging speed and system instability, restricting acquisition volume.

Purpose of the Study:

To present techniques for accelerating FIB-SEM image acquisition and enhancing system reliability.
To enable large-volume, high-resolution 3D imaging for applications like connectomics.

Main Methods:

Development of novel techniques to improve imaging speed and long-term system stability.
Continuous imaging of large volumes exceeding 10^6 µm³.

Main Results:

Significantly accelerated image acquisition and improved FIB-SEM reliability.
Sustained system operation for months, generating large, continuously imaged volumes.
Demonstrated high-resolution data from mammalian neural tissue, Drosophila brain, and Chlamydomonas reinhardtii.

Conclusions:

The enhanced FIB-SEM technique overcomes previous limitations, enabling large-scale, high-resolution 3D reconstruction.
This advancement is crucial for connectomics, aiding neuronal process tracing and reducing manual proofreading.
The technique provides powerful insights for both connectomics and cell biology research.