Gas cluster ion beam SEM for imaging of large tissue samples with 10 nm isotropic resolution

• Published in: Nature methods Vol. 17; no. 1; pp. 68 - 71
• Main Authors: Hayworth, Kenneth J., Peale, David, Januszewski, Michał, Knott, Graham W., Lu, Zhiyuan, Xu, C. Shan, Hess, Harald F.
• Format: Journal Article
• Language: English
• Published: New York Nature Publishing Group US 01.01.2020; Nature Publishing Group
• Subjects: 631/1647/328/1649; 631/1647/334/1582/715; 631/1647/334/1874/345; 631/378; 631/378/2597; Animals; Bioinformatics; Biological Microscopy; Biological Techniques; Biomedical and Life Sciences; Biomedical Engineering/Biotechnology; Brain - ultrastructure; Brief Communication; Cerebral Cortex - ultrastructure; Clusters; Drosophila melanogaster; Electron microscopy; Image Processing, Computer-Assisted - methods; Imaging, Three-Dimensional - methods; Ion beams; Life Sciences; Male; Mice; Mice, Inbred C57BL; Microscopy, Electron, Scanning - methods; Proteomics; Reliability aspects; Scanning electron microscopy; Tissue Fixation
• ISSN: 1548-7091; 1548-7105; 1548-7105
• DOI: 10.1038/s41592-019-0641-2

We demonstrate gas cluster ion beam scanning electron microscopy (SEM), in which wide-area ion milling is performed on a series of thick tissue sections. This three-dimensional electron microscopy technique acquires datasets with <10 nm isotropic resolution of each section, and these can then be stitched together to span the sectioned volume. Incorporating gas cluster ion beam SEM into existing single-beam and multibeam SEM workflows should be straightforward, increasing reliability while improving z resolution by a factor of three or more. An alternative to focused ion beam scanning electron microscopy (FIB-SEM), gas cluster ion beam scanning electron microscopy (GCIB-SEM) is compatible with large tissue samples while achieving similar isotropic resolution.