Multi-beam two-photon imaging of fast Ca2+ signals in the Langendorff mouse heart

Although the role of calcium (Ca(2+)) in excitation-contraction coupling in the heart can be comprehensively studied at the cellular level, propagation of Ca(2+) signals intercellularly requires tissue-based investigations. To access cells below the epicardium, an optical-sectioning technique is necessary. Multi-photon microscopy allows reliable imaging for penetration to depths of up to 0.5 mm. Here, we provide a protocol that uses multibeam two-photon microscopy for measuring Ca(2+) signals in a Langendorff-perfused mouse heart.