Instantaneous isotropic volumetric imaging of fast biological processes

To capture highly dynamic biological processes at cellular resolution is a recurring challenge in biology. Here we show that combining selective-volume illumination with simultaneous acquisition of orthogonal light fields yields three-dimensional images with high, isotropic spatial resolution and a significant reduction of reconstruction artefacts, thereby overcoming current limitations of light-field microscopy implementations. We demonstrate medaka heart and blood flow imaging at single-cell resolution and free of motion artefacts at volume rates of up to 200 Hz. Iso-LFM enables rapid, instantaneous volumetric imaging of biological processes with isotropic and improved resolution by simultaneously capturing orthogonal light fields.