Two photon imaging of calcium responses in murine Purkinje neurons

Purkinje neurons (PNs) are an important component of the motor learning and coordination circuit and are affected in spino-cerebellar ataxias. Maintaining healthy PNs in cerebellar slices and recording their Ca²⁺ transients can be challenging. Here, we describe a protocol for measuring Ca²⁺ transients in PNs from adult mice, including problem-solving tips. This protocol can be used to measure neuronal excitability and agonist-mediated Ca²⁺ signaling in cerebellar slices expressing a genetic Ca²⁺ reporter in all PNs, thus improving yield of data. For complete details on the use and execution of this profile, please refer to Dhanya and Hasan (2021).