FIP Seminar - "Mapping cellular function with 3D single-molecule tracking and super-resolution microscopy"

TCellular function is governed by the molecular organization and interactions at the nanoscale. In this talk I will demonstrate our recent developments for improved 3D single-molecule tracking of dynamics and super-resolution imaging of nanoscale structures throughout mammalian cells and showcase applications of our approaches for cellular imaging. I will describe our developments of light sheet microscopy platforms that reduce fluorescence background, photobleaching, and the risk of photodamaging sensitive samples. Combined with point spread function (PSF) engineering for nanoscale localization of individual molecules in 3D, deep learning for analysis of overlapping emitters, and a novel 3D nanoprinted microfluidic chip for environmental control, our platforms offer whole-cell multi-target 3D single-molecule super-resolution imaging with improved accuracy, precision, and imaging speed. Next, I will demonstrate how we integrate the optical sectioning capabilities of light sheet illumination with uniform, flat-field epi- and TIRF illumination to achieve more precise and accurate quantitation of single-molecule data. I will also demonstrate novel long axial-range double-helix PSFs and show that they offer stitching-free, 3D super-resolution imaging of whole mammalian cells, simplifying the experimental and analysis procedures for obtaining volumetric nanoscale structural information. Furthermore, I will show that deep learning-based analysis drastically improves the achievable imaging speed and resolution with these PSFs. Finally, I will describe our recent developments and applications of dCas9-based labels for flexible and long-term tracking of endogenous, non-repetitive genomic loci in live human cells with excellent spatiotemporal resolution. These imaging approaches are versatile and can be utilized to study molecular dynamics, nanoscale structures, and molecular mechanisms to address a broad range of chemical, biological, and biomedical questions related to cellular function and pathogenesis.

Dr. Gustavsson joined the faculty at Rice University in 2020 as a CPRIT Scholar in Cancer Research and the Norman Hackerman-Welch Young Investigator Chair. At Rice, she founded and serve as Director of the Center for Nanoscale Imaging Sciences. Her research group strives to gain detailed information about cellular nanoscale structures, dynamics, and molecular mechanisms by designing and applying innovative and versatile optical imaging tools.