STAR Protocols (Dec 2021)

Protocol for cortical-wide field-of-view two-photon imaging with quick neonatal adeno-associated virus injection

  • Ikumi Oomoto,
  • Hiroyuki Uwamori,
  • Chie Matsubara,
  • Maya Odagawa,
  • Midori Kobayashi,
  • Kenta Kobayashi,
  • Keisuke Ota,
  • Masanori Murayama

Journal volume & issue
Vol. 2, no. 4
p. 101007

Abstract

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Summary: We recently established a simple and versatile adeno-associated virus (AAV) induction approach that enables dense (>90% labeled neurons) and cortical-wide Ca2+ sensor expression. Here, we describe the stepwise protocol for neonatal AAV injection of a Ca2+ sensor. We also detail the steps for subsequent craniotomy to generate a chronic cranial window, followed by wide-field two-photon Ca2+ imaging in an awake mouse. This protocol serves as an alternative to the use of transgenic animals and offers translatable options for cortical-wide experiments.For complete details on the use and execution of this protocol, please refer to Ota et al. (2021).

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