"This award is funded under the American Recovery and Reinvestment Act of 2009 (Public Law 111-5)."
0934860 Vainrub
Light microscopy is the major biological research technology that enabled modern knowledge of structure and function of biological cells. With the discovery of super-resolution light microscopy in the late 1990s the size of observable features diminished twenty times to as small as ten nanometers, promising to revolutionize sub-cellular and molecular biology research. Super-resolution microscopy is still in its infancy. Specifically, imaging speeds are below 1 frame/s. The research objective of this proposal is the development of a novel microscopy platform that combines spatial super-resolution in all three dimensions with high imaging speed of 5000 frames/s to enable study of fast intracellular events. The principle of the method is based on simultaneous illumination of the object by about a hundred thousand narrow light spots, each focused to the diffraction-limited size. The illumination pattern is generated by a computer-controlled digital micro-mirror device (DMD); the pattern's quality satisfies the rigid super-resolution conditions as tested by preliminary experiments. The super-resolution image will be reconstructed using 9-25 frames recorded for different illuminations. Theoretically, in linear mode the 3D resolution enhancement is two-fold compared to the classical diffraction limit. In non-linear mode of saturated fluorescence further resolution enhancement occurs with no theoretical limit. This supreme 3D imaging capability will be due to the super-resolution in axial direction and low out-of-focus light. The developed technique will be widely applicable to the study of the structural organization and dynamic processes in living cells, in particular in the area of mitochondria research.
