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Jan Huisken (Max Planck Institute of Molecular Cell Biology and Genetics, Dresden, Germany)
Selective plane illumination microscopy (SPIM) is a fluorescence microscopy technique that uses a focused light-sheet to illuminate the specimen from the side. SPIM achieves excellent resolution at high penetration depths while being minimally invasive at the same time (Huisken & Stainier, Development, 2009). SPIM offers a number of advantages over established techniques such as strongly reduced photo-bleaching, much higher dynamic range, and much higher acquisition speed. The extraordinary speed of the microscope allows us to image the zebrafish heart in full detail, in three dimensions over time. Using a dual transgenic flk1:GFP and cmlc2:dsred we have been able to image the process of atrial contraction in the embryonic zebrafish heart with high spatial and temporal resolution. Individual movies of the zebrafish heart are taken at different positions. They are registered and synchronized by aligning tilted views in spatial and temporal dimensions. The resulting multi-dimensional data is visualized in a variety of different ways to analyze the mechanical force transmission from myocardium to endocardium. The result is a multi-color 3D movie of the cardiac function in real time.