Volumetric super-resolution imaging by serial ultrasectioning and stochastic optical reconstruction microscopy in mouse neural tissue

STAR Protoc. 2021 Nov 24;2(4):100971. doi: 10.1016/j.xpro.2021.100971. eCollection 2021 Dec 17.

Abstract

Here, we present a protocol for collecting large-volume, four-color, single-molecule localization imaging data from neural tissue. We have applied this technique to map the location and identities of chemical synapses across whole cells in mouse retinae. Our sample preparation approach improves 3D STORM image quality by reducing tissue scattering, photobleaching, and optical distortions associated with deep imaging. This approach can be extended for use on other tissue types enabling life scientists to perform volumetric super-resolution imaging in diverse biological models. For complete details on the use and execution of this protocol, please refer to Sigal et al. (2015).

Keywords: Cell Biology; Microscopy; Neuroscience.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Female
  • Imaging, Three-Dimensional / methods*
  • Immunohistochemistry / methods*
  • Male
  • Mice
  • Retina* / chemistry
  • Retina* / cytology
  • Retina* / diagnostic imaging
  • Single Molecule Imaging / methods*
  • Synapses / chemistry*