The Centre for Optophysiology is a core facility jointly established by the University of Toronto, the Lunenfeld Tanenbaum Research Institute at Mount Sinai Hospital and SickKids Hospital. State-of-the-art microscopy systems available include a dual multiphoton (MP) microscope with spatial light modulator, a Stimulated Emission Depletion (STED) microscope for super resolution imaging and an in vivo MP microscope with Neurotar homecage for imaging awake mice. The Centre is located at two sites that are <500 m apart in the heart of Toronto’s Discovery District: the Physiology Department at the University of Toronto and The Centre for Phenogenomics (TCP). The equipment can be used to image preparations ranging from cultured cells or tissue slices to whole animals and can be used in combination with electrophysiology. The Centre is open to researchers not only from the collaborating institutions, but also from the wider Toronto area.