STAR Protocols (Dec 2020)

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

  • Georg Kislinger,
  • Helmut Gnägi,
  • Martin Kerschensteiner,
  • Mikael Simons,
  • Thomas Misgeld,
  • Martina Schifferer

Journal volume & issue
Vol. 1, no. 3
p. 100232

Abstract

Read online

Summary: 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