Two photon imaging of calcium responses in murine Purkinje neurons

Purkinje neurons (PNs) in vertebrate cerebella are required for motor learning and motor co-ordination. Dysregulation of intracellular calcium signaling in PNs can be a causative pathogenic factor for Spino-cerebellar Ataxias. Our protocol describes a method for measuring Ca2+ transients in PNs from adult mice. This protocol can measure neuronal excitability and agonist mediated Ca2+ signaling in cerebellar slices. As maintaining healthy PNs in cerebellar slices and recording their Ca2+ transients are challenging, we provide detailed descriptions and problem-solving tips.