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

STAR Protoc. 2021 Dec 10;2(4):101007. doi: 10.1016/j.xpro.2021.101007. eCollection 2021 Dec 17.

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).

Keywords: Microscopy; Model Organisms; Neuroscience.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Calcium / metabolism
  • Cerebral Cortex / diagnostic imaging*
  • Craniotomy
  • Dependovirus / genetics*
  • Female
  • Genetic Vectors / administration & dosage
  • Genetic Vectors / genetics
  • Injections
  • Male
  • Mice
  • Optical Imaging / methods*
  • Skull / surgery

Substances

  • Calcium