Project Abstract Light-sheet fluorescence microscopy (LSFM), through its gentle, efficient, and fast 3D imaging capacity, has tremendous potential in biological, biomedical, and translational applications. However, LSFM has fragmented into a myriad of specialized instruments, each optimized for a specific class of samples and imaging regimes. As such, the widespread use and ultimate impact of LSFM has been curtailed by its lack of adaptability. Here we propose a universal LSFM platform that can be adapted to a wide range of applications, ranging from sensitive live cell imaging to imaging organs and tissues that have been rendered transparent by different clearing techniques. Further, we will improve the spatial and temporal resolution, as well as the volumetric coverage of LSFM. These additional improvements will be packaged in modules, which can be integrated into our proposed platform on demand. To demonstrate its biomedical potential, we will image human cardiomyocytes, both live and with super-resolution microscopy, to evaluate the performance of emerging cardiac therapies on a molecular level. In its most sensitive configuration, our LSFM platform will be able to study spatiotemporal patterns of calcium signaling, in health and disease, which we will analyze with statistical times series analysis methods. Overall, our proposal will increase the capabilities and accessibility of LSFM, and as such will spur three-dimensional imaging in biological and biomedical research.

Project Narrative We propose a versatile and flexible light-sheet fluorescence microscopy (LSFM) platform, which will be able to perform a wide range of imaging applications, ranging from single cells to cleared tissues. LSFM are typically highly specialized instruments, but we hypothesize that versatility is needed to realize the full potential of LSFM in the life sciences. To study the efficacy of therapies, we will image calcium signaling and cytoskeletal structure in human derived cardiomyocytes, highlighting the adaptability and potential of our instrument in biomedical research.