A pipeline for STED super-resolution imaging and Imaris analysis of nanoscale synapse organization in mouse cortical brain slices

STAR Protoc. 2023 Dec 15;4(4):102707. doi: 10.1016/j.xpro.2023.102707. Epub 2023 Nov 9.

Abstract

Advances in super-resolution imaging enable us to delve into its intricate structural and functional complexities with unprecedented detail. Here, we present a pipeline to visualize and analyze the nanoscale organization of cortical layer 1 apical dendritic spines in the mouse prefrontal cortex. We describe steps for brain slice preparation, immunostaining, stimulated emission depletion super-resolution microscopy, and data analysis using the Imaris software package. This protocol allows the study of physiologically relevant brain circuits implicated in neuropsychiatric disorders.

Keywords: Cell Biology; Microscopy; Neuroscience.

MeSH terms

  • Animals
  • Brain / diagnostic imaging
  • Central Nervous System
  • Dendrites*
  • Mice
  • Microscopy* / methods
  • Synapses