Hayworth, Kenneth J.Xu, C. ShanLu, ZhiyuanKnott, Graham W.Fetter, Richard D.Tapia, Juan CarlosLichtman, JeffHess, Harald F.2015-11-032015Hayworth, Kenneth J., C. Shan Xu, Zhiyuan Lu, Graham W. Knott, Richard D. Fetter, Juan Carlos Tapia, Jeff W. Lichtman, and Harald F. Hess. 2015. “Ultrastructurally-smooth thick partitioning and volume stitching for larger-scale connectomics.” Nature methods 12 (4): 319-322. doi:10.1038/nmeth.3292. http://dx.doi.org/10.1038/nmeth.3292.1548-7091http://nrs.harvard.edu/urn-3:HUL.InstRepos:23474101FIB-SEM has become an essential tool for studying neural tissue at resolutions below 10×10×10 nm, producing datasets superior for automatic connectome tracing. We present a technical advance, ultrathick sectioning, which reliably subdivides embedded tissue samples into chunks (20 µm thick) optimally sized and mounted for efficient, parallel FIB-SEM imaging. These chunks are imaged separately and then ‘volume stitched’ back together, producing a final 3D dataset suitable for connectome tracing.en-USJournal Article2015-11-0310.1038/nmeth.3292