Serial-section electron microscopy using automated tape-collecting ultramicrotome (ATUM)

Methods Cell Biol. 2019:152:41-67. doi: 10.1016/bs.mcb.2019.04.004. Epub 2019 Jun 8.

Abstract

The Automated Tape-Collecting Ultramicrotome (ATUM) is a tape-reeling device that is placed in a water-filled diamond knife boat to collect serial sections as they are cut by a conventional ultramicrotome. The ATUM can collect thousands of sections of many different shapes and sizes, which are subsequently imaged by a scanning electron microscope. This method has been used for large-scale connectomics projects of mouse brain, and is well suited for other smaller-scale studies of tissues, cells, and organisms. Here, we describe basic procedures for preparing a block for ATUM sectioning, handling of the ATUM, tape preparation, post-treatment of sections, and considerations for mapping, imaging, and aligning the serial sections.

Keywords: 3D EM; ATUM; SEM; Serial section EM; Tape collector.

MeSH terms

  • Animals
  • Brain / physiology
  • Image Processing, Computer-Assisted / methods
  • Imaging, Three-Dimensional / methods
  • Mice
  • Microscopy, Electron, Scanning / methods*
  • Microtomy / methods*