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Ultrathick (Hot Knife) Sectioning

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Ultrathick (Hot Knife) Sectioning
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In recent years, we have developed a technical advance, ultrathick (hot knife) sectioning, which reliably subdivides embedded tissue samples into chunks (20 μm thick) optimally sized and mounted for parallel FIB-SEM imaging. These chunks are imaged separately and then 'volume stitched' back together, producing a final three-dimensional data set suitable for connectome tracing. The following two movies illustrate the mechanics of hot knife sectioning and the result of ultrathick sectioning and volume stitching.