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

Article · Xu CS, Hayworth KJ, Lu Z, Grob P, Hassan AM, García-Cerdán JG, Niyogi KK, Nogales E, Weinberg RJ, Hess HF

- Archive id: doi:10.7554/elife.25916
- Published: 2017-05-13
- Venue: eLife
- DOI: https://doi.org/10.7554/elife.25916
- PMID: 28500755
- PMCID: PMC5476429
- Citations: 339
- Source URL: https://doi.org/10.7554/elife.25916
- PDF: https://europepmc.org/articles/PMC5476429/pdf
- Provenance: europepmc

## Abstract

Focused Ion Beam Scanning Electron Microscopy (FIB-SEM) can automatically generate 3D images with superior z-axis resolution, yielding data that needs minimal image registration and related post-processing. Obstacles blocking wider adoption of FIB-SEM include slow imaging speed and lack of long-term system stability, which caps the maximum possible acquisition volume. Here, we present techniques that accelerate image acquisition while greatly improving FIB-SEM reliability, allowing the system to operate for months and generating continuously imaged volumes \> 10 6 µm 3 . These volumes are large enough for connectomics, where the excellent z resolution can help in tracing of small neuronal processes and accelerate the tedious and time-consuming human proofreading effort. Even higher resolution can be achieved on smaller volumes. We present example data sets from mammalian neural tissue, Drosophila brain, and Chlamydomonas reinhardtii to illustrate the power of this novel high-resolution technique to address questions in both connectomics and cell biology.

_Abstract reproduced under CC-BY (article license as recorded by Europe PMC; abstract reproduced unmodified apart from markup removal)._

Topics: [Connectomics & Brain Mapping](https://space-kitty.com/topics/connectomics)

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