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

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).

Automated summary

This AAV induction approach enables efficient neuronal labeling with Ca2+ sensors and allows for calcium imaging experiments throughout the entire cortical area in mice.