Brent Segelke | 19-LW-048
We need a better understanding of infectious disease to safeguard human health, the economy, and global security. Modern omics techniques hold the promise of providing a comprehensive understanding of the molecular mechanisms of life, including pathogenesis from infectious disease, but there is a significant gap in annotation of gene function. Recent innovations in fluorescence microscopy for live-cell imaging and genetic engineering make it possible to determine the temporal correlation between molecular events and cellular behavior. This provides new insight into molecular mechanisms of pathogenesis, which will provide new therapeutic targets or novel countermeasure strategies.
The goal of our project was to (1) develop a lattice light sheet fluorescence microscope for long-time-course, live-cell imaging experiments; (2) develop the reagents and cell lines needed to monitor molecular events while bacteria infect mammalian immune cells; and (3) demonstrate that we could capture molecular events during an infection. We fully commissioned the Lawrence Livermore National Laboratory lattice light sheet microscope and conducted initial, live-cell studies with immune cells and pathogenic bacteria. Our work showed that long-time-course, live-cell imaging has tremendous potential to help elucidate molecular mechanisms of host-pathogen interactions and to help annotate gene function, which would establish a basis for new countermeasures.
This project directly impacts Livermore's mission focus area in weapons of mass destruction (WMD) threat reduction, providing unique capabilities, expertise, and innovative solutions applicable to biothreat agents and emerging threats. This project directly supported the Laboratory's bioscience and bioengineering core competency, providing a capability and team delivering physiologically relevant, high-fidelity, in vitro and in silico tools to interrogate biological systems across many orders of magnitude of length and timescales. These new capabilities dramatically enhance our ability to interrogate cellular mechanisms and the interaction among cells. In addition, this project could impact the Director's Initiative in predictive biology by providing state-of-the-art, precision experimental measurements in cellular biology, microbiology, and molecular biology.