PT - JOURNAL ARTICLE AU - Anna-Karin Gustavsson AU - Petar N. Petrov AU - Maurice Y. Lee AU - Yoav Shechtman AU - W. E. Moerner TI - 3D single-molecule super-resolution microscopy with a tilted light sheet AID - 10.1101/135699 DP - 2017 Jan 01 TA - bioRxiv PG - 135699 4099 - http://biorxiv.org/content/early/2017/08/14/135699.short 4100 - http://biorxiv.org/content/early/2017/08/14/135699.full AB - Tilted light sheet microscopy with 3D point spread functions (TILT3D) combines a novel, tilted light sheet illumination strategy with long axial range point spread functions (PSFs) for low-background, 3D super-localization of single molecules as well as 3D super-resolution imaging in thick cells. Because the axial positions of the single emitters are encoded in the shape of each single-molecule image rather than in the position or thickness of the light sheet, the light sheet need not be extremely thin. TILT3D is built upon a standard inverted microscope and has minimal custom parts. The result is simple and flexible 3D super-resolution imaging with tens of nm localization precision throughout thick mammalian cells. We validated TILT3D for 3D super-resolution imaging in mammalian cells by imaging mitochondria and the full nuclear lamina using the double-helix PSF for single-molecule detection and the recently developed Tetrapod PSFs for fiducial bead tracking and live axial drift correction.