Mapping cellular function with 3D single molecule tracking and super-resolution microscopy
Abstract
Cellular 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 superresolution 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.
Biography
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 serves 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. Dr. Gustavsson received her Ph.D. in Physics from the University of Gothenburg, Sweden. Her graduate work focused on studying rhythms and dynamic responses in single cells by combining and optimizing techniques such as fluorescence microscopy, optical tweezers, and microfluidics. Upon completion of her graduate work, Dr. Gustavsson joined the group of Nobel Laureate W. E. Moerner at Stanford University as a Postdoctoral Fellow. Her research focused on the development and application of 3D single-molecule super-resolution microscopy for cellular imaging and included the implementation of light sheet illumination for optical sectioning of mammalian cells. Her work has been recognized with multiple honors, awards, and fellowships, most notably the FEBS Journal Richard Perham Prize, the 3-year Swedish Research Council International Postdoctoral Fellowship, the PicoQuant Young Investigator Award, the NIH K99/R00 Pathway to Independence Award, the CPRIT Recruitment of First-Time Tenure-Track Faculty Members Award, the Scialog: Advancing Bioimaging Fellowship, the Edward S. and Fofo Lewis Chemistry Research Award, and the NSF CAREER Award
Click here to register for in-person attendance (lunch will be served)
