Light disc microscopy

C53-28a-271090Biological systems function across scales, with operations at nanoscales coming together to form complex macroscale communities. Thus, resolving interactions and tracking metabolic processes within living systems requires multi-scale imaging to visualize cells at the community, cellular, subcellular, and single-molecule scales. Although techniques like light sheet fluorescence microscopy can achieve this multi-scale imaging, light sheets illuminate only one side of samples, resulting in shadowing artifacts that can obscure the field of view and prevent proper interpretation of the larger system. This Phase II SBIR/STTR project will develop a novel parabolic mirror-based imaging system that is compatible with existing inverted microscopes and that eliminates shadowing across scales—at both high and low resolution—to enable accurate interpretation of any specimen. The project objectives are to build a second-generation prototype and test its performance on a range of samples, to initiate alpha testing with collaborators, and to investigate the use with structured illumination with this system to allow for super-resolution imaging. The ability to visualize microbial communities without shadowing at both low magnification and super-resolution will open up new experimental space for investigating the complex energetic relationships in microbial systems. Furthermore, in the long term, the radical design change made possible by this technology will provide a platform for further innovation, enabling the development of completely new imaging techniques and expanding what is currently possible in the field of live-cell fluorescence microscopy.