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

ABSTRACT: 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).

SUBMITTER: Oomoto I

PROVIDER: S-EPMC8672102 | biostudies-literature | 2021 Dec

REPOSITORIES: biostudies-literature

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

Oomoto Ikumi I Uwamori Hiroyuki H Matsubara Chie C Odagawa Maya M Kobayashi Midori M Kobayashi Kenta K Ota Keisuke K Murayama Masanori M

STAR protocols 20211210 4

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 us ...[more]

PMID: 34950887

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