Imaging adultC. eleganslive using light-sheet microscopy

Live observation of biological phenomena in the context of living organisms can provide important insights in the mechanisms of these phenomena. However, the spatially complex and dynamic physiology of multicellular organisms can be a challenging environment to make observations with fluorescence microscopy. Due to the illumination of out‐of‐focus planes, confocal and particularly widefield fluorescence microscopy suffer from low signal‐to‐background ratio (SBR), photo toxicity and bleaching of fluorescent probes. In light‐sheet microscopy (LSM), solely the focal plane of the detection objective is illuminated, minimising out‐of‐focus fluorescence and photobleaching, thereby enhancing SBR, allowing for low laser intensities and longer acquisition periods. Here we present a straightforward light‐sheet microscope with a 1.0‐NA detection objective and a fast sample‐positioning stage that allows for four degrees of freedom. By imaging the sensory cilia and nervous system of living young adult C. elegans, we demonstrate that the instrument is well suited for relatively fast, volumetric imaging of larger (hundreds of micrometres cubed) living samples. These experiments demonstrate that such an instrument provides a valuable addition to commonly used widefield and confocal fluorescence microscopes.