To obtain a complete picture of subcellular structures, cells must be imaged with high resolution in all three dimensions (3D). In this talk, I will present tilted light sheet microscopy with 3D point spread functions (TILT3D), an imaging platform that combines a novel, tilted light sheet illumination strategy with engineered 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. Here the axial positions of the single molecules are encoded in the shape of the PSF rather than in the position or thickness of the light sheet. 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. We think that TILT3D in the future will become an important tool not only for 3D super-resolution imaging, but also for live whole-cell single-particle and single-molecule tracking.
Dr. Anna-Karin Gustavsson is a postdoctoral fellow in the Moerner Lab at the Department of Chemistry at Stanford University, and she also holds a postdoctoral fellowship from the Karolinska Institute in Stockholm, Sweden. Her research is focused on the development and application of 3D single-molecule super-resolution microscopy for cell imaging, and includes the implementation of light sheet illumination for optical sectioning. She has a background in physics and received her PhD in Physics in 2015 from the University of Gothenburg, Sweden. Her PhD project was focused on studying dynamic responses in single cells by combining and optimizing techniques such as fluorescence microscopy, optical tweezers, and microfluidics. Dr. Gustavsson has received several awards, most notably the FEBS Journal Richard Perham Prize for Young Scientists in 2012 and the PicoQuant Young Investigator Award in 2018.