TY - JOUR TI - Enhanced FIB-SEM systems for large-volume 3D imaging AU - Xu, C Shan AU - Hayworth, Kenneth J AU - Lu, Zhiyuan AU - Grob, Patricia AU - Hassan, Ahmed M AU - García-Cerdán, José G AU - Niyogi, Krishna K AU - Nogales, Eva AU - Weinberg, Richard J AU - Hess, Harald F A2 - Nathans, Jeremy VL - 6 PY - 2017 DA - 2017/05/13 SP - e25916 C1 - eLife 2017;6:e25916 DO - 10.7554/eLife.25916 UR - https://doi.org/10.7554/eLife.25916 AB - 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 > 106 µm3. 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. KW - connectomics KW - 3D cellular structure KW - electron microscopy KW - imaging KW - FIB JF - eLife SN - 2050-084X PB - eLife Sciences Publications, Ltd ER -