Super-resolution, structured illuminationmicroscope (SIM) for biomedical research - Project Summary: Our understanding of fundamental cell biological processes is both driven by and limited by our ability to visualize the organization of structures and molecules within cells and tissues. The development of super- resolution fluorescence imaging methods that circumvent the resolution limits of conventional light microscopy, achieving lateral resolution of <100 nm, have transformed cell biological research. Access to super-resolution fluorescence imaging technology is now essential for scientists wishing to push the frontiers of cell biological research. This proposal requests funds to replace Stanford University’s Cell Sciences Imaging Facility’s 11-year-old, obsolete OMX structured illumination, super resolution microscope with an Elyra 7 lattice- SIM2 super-resolution imaging system (Zeiss, Inc.). This state-of-the-art, super-resolution microscope can achieve both lateral and axial resolution at greater than twice the diffraction limit of conventional light microscopy and is capable of multichannel super-resolution far beyond the cover glass. The fast-imaging capability of Elyra 7 lattice SIM2 is also designed to overcome speed limitations enabling 3D live-cell super resolution imaging. This advanced imaging system will be a shared resource, located in a well-established, multi-user microscopy facility at Stanford: The Cell Sciences Imaging Facility. The requested Elyra 7 lattice SIM2 imaging system will support research projects from eight researchers, all of which are NIH funded. These projects investigate a wide range of topics, including: meiotic chromosome segregation and genetic recombination (Villeneuve); signaling and vesicular transport in primary cilia (Jackson), molecular mechanisms of myelin wrapping in neurons (Zuchero); maintenance and regulation of genomic stability (Cimprich and Chistol), spatial organization of cell-cell junctions (Dunn); the role of the extra cellular matrix in cell invasion and migration (Chaudhuri); and the role human calcineurin signaling pathways in cellular processes (Cyert). These studies investigate critical functional and structural questions regarding fundamental cell biological processes and cover NIH research areas with implications for diverse aspects of human health and disease, including cancer, birth defects, obesity, immunity, as well as neurodegenerative disease. All these projects require multi-channel super-resolution imaging and simultaneous multi-channel fast imaging; this combination of capabilities is most effectively provided by the requested Elyra 7 lattice SIM2 instrument.
