ATUM-FIB microscopy for targeting and multiscale imaging of rare events in mouse cortex

STAR Protoc. 2020 Dec 16;1(3):100232. doi: 10.1016/j.xpro.2020.100232. eCollection 2020 Dec 18.


Here, we describe a detailed workflow for ATUM-FIB microscopy, a hybrid method that combines serial-sectioning scanning electron microscopy (SEM) with focused ion beam SEM (FIB-SEM). This detailed protocol is optimized for mouse cortex samples. The main processing steps include the generation of semi-thick sections from sequentially cured resin blocks using a heated microtomy approach. We demonstrate the different imaging modalities, including serial light and electron microscopy for target recognition and FIB-SEM for isotropic imaging of regions of interest. For complete details on the use and execution of this protocol, please refer to Kislinger et al. (2020).

Keywords: Microscopy; Model organisms; Neuroscience.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Cerebral Cortex / diagnostic imaging*
  • Cerebral Cortex / metabolism
  • Image Processing, Computer-Assisted / methods
  • Imaging, Three-Dimensional / methods
  • Mice
  • Microscopy, Electron, Scanning / methods*
  • Microtomy / instrumentation
  • Microtomy / methods*
  • Workflow