Oblique plane microscope for mesoscopic imaging of freely moving organisms with cellular resolution

Several important questions in biology require non-invasive and three-dimensional imaging techniques with an appropriate spatiotemporal resolution that permits live organisms to move in an unconstrained fashion over an extended field-of-view. While selective-plane illumination microscopy (SPIM) has emerged as a powerful method to observe live biological specimens at high spatio-temporal resolution, typical implementations often necessitate constraining sample mounting or lack the required volumetric speed. Here, we report on an open-top, dual-objective oblique plane microscope (OPM) capable of observing millimeter-sized, freely moving animals at cellular resolution. We demonstrate the capabilities of our mesoscopic OPM (MesOPM) by imaging the behavioral dynamics of the sea anemone Nematostella vectensis over 1.56 x 1.56 x 0.25 mm at 1.5 x 2.8 x 5.3 mu m resolution and 0.5 Hz volume rate.

Automated summary

Non-invasive and three-dimensional imaging techniques with appropriate spatiotemporal resolution are necessary to observe freely moving organisms in biology. The open-top, dual-objective oblique plane microscope (OPM) presented in this study allows for cellular resolution imaging of sea anemone behavior. The MesOPM demonstrated the capability to image the behavioral dynamics of the sea anemone Nematostella vectensis over a volume of 1.56 x 1.56 x 0.25 mm at a resolution of 1.5 x 2.8 x 5.3 μm and a rate of 0.5 Hz.