Protocol to image and analyze hippocampal network dynamics in non-anesthetized mouse pups

Summary: Two-photon calcium imaging is a powerful technique that has revolutionized our understanding of how neural circuit dynamics supports different behaviors and cognitive processes. However, performing imaging during development remains challenging. Here, we provide a protocol to image CA1 neurons in mouse pups as well as a pipeline of analysis to analyze and share the data. We describe steps for intracerebroventricular injection, cranial window surgery, two-photon calcium imaging, and analysis of imaging data.For complete details on the use and execution of this protocol, please refer to Dard et al.1 and Denis et al.2 : Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics.