Two-photon in vivo imaging of dendritic spines in the mouse cortex using a thinned-skull preparation

Xinzhu Yu, Yi Zuo

Research output: Contribution to journalArticlepeer-review

Abstract

In the mammalian cortex, neurons form extremely complicated networks and exchange information at synapses. Changes in synaptic strength, as well as addition/removal of synapses, occur in an experience-dependent manner, providing the structural foundation of neuronal plasticity. As postsynaptic components of the most excitatory synapses in the cortex, dendritic spines are considered to be a good proxy of synapses. Taking advantages of mouse genetics and fluorescent labeling techniques, individual neurons and their synaptic structures can be labeled in the intact brain. Here we introduce a transcranial imaging protocol using two-photon laser scanning microscopy to follow fluorescently labeled postsynaptic dendritic spines over time in vivo. This protocol utilizes a thinned-skull preparation, which keeps the skull intact and avoids inflammatory effects caused by exposure of the meninges and the cortex. Therefore, images can be acquired immediately after surgery is performed. The experimental procedure can be performed repetitively over various time intervals ranging from hours to years. The application of this preparation can also be expanded to investigate different cortical regions and layers, as well as other cell types, under physiological and pathological conditions.

Original languageEnglish (US)
Article numbere51520
JournalJournal of Visualized Experiments
Issue number87
DOIs
StatePublished - May 12 2014
Externally publishedYes

Keywords

  • Dendritic spine
  • Imaging
  • In vivo
  • Issue 87
  • Mouse cortex
  • Neuroscience
  • Thinned-skull
  • Two-photon microscopy

ASJC Scopus subject areas

  • General Neuroscience
  • General Chemical Engineering
  • General Biochemistry, Genetics and Molecular Biology
  • General Immunology and Microbiology

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