In vivo two-photon imaging of sensory-evoked dendritic calcium signals in cortical neurons

Hongbo Jia, Nathalie L. Rochefort, Xiaowei Chen, Arthur Konnerth*

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review

Abstract

Neurons in cortical sensory regions receive modality-specific information through synapses that are located on their dendrites. Recently, the use of two-photon microscopy combined with whole-cell recordings has helped to identify visually evoked dendritic calcium signals in mouse visual cortical neurons in vivo. The calcium signals are restricted to small dendritic domains ('hotspots') and they represent visual synaptic inputs that are highly tuned for orientation and direction. This protocol describes the experimental procedures for the recording and the analysis of these visually evoked dendritic calcium signals. The key points of this method include delivery of fluorescent calcium indicators through the recording patch pipette, selection of an appropriate optical plane with many dendrites, hyperpolarization of the membrane potential and two-photon imaging. The whole protocol can be completed in 5–6 h, including 1–2 h of two-photon calcium imaging in combination with stable whole-cell recordings.
Original languageEnglish
Pages (from-to)28-35
Number of pages8
JournalNature Protocols
Volume6
Issue number1
DOIs
Publication statusPublished - Jan 2011

Keywords

  • NEOCORTICAL PYRAMIDAL NEURONS
  • SINGLE-CELL ELECTROPORATION
  • MAMMALIAN BRAIN
  • NEURAL ACTIVITY
  • VISUAL-CORTEX
  • SPINES
  • INDICATORS
  • RECORDINGS
  • DYNAMICS

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