REAL-TIME SINGLE MOLECULE LIVE CELL MEASUREMENT OF TRANSCRIPTION BY 3D ORBITAL TRACKING FLUORESCENCE CROSS-CORRELATION SPECTROSCOPY

Mechanisms of transcription and translation take the information encoded in the genome and make it "work" in cells, through the production of proteins defined by nucleic acid coding regions. This involves the coordination of many multi-subunit complexes about which most knowledge is inferred from ensemble and/or in vitro assays, giving a detailed but static picture. How these macromolecular machines coordinate in living cells remains unknown but recent advances in the application of fluctuation analysis to time-resolved multi-color fluorescence imaging can now give an unprecedented level of dynamic in vivo information.

My talk will describe recent applications of 3D Orbital Tracking and Fluorescence Cross-Correlation Spectroscopy to the study of active genes in living cells and outline possible future applications. Orbital tracking provides faster sampling and longer measurements than traditional microscopy while minimizing photobleaching. Using these methods, we are able to begin to understand and model the living genome.

Matthew L. Ferguson was trained in physics receiving a B.S. in physics and mathematics from Texas Christian University and a Ph.D. in physics from the University of Maryland, College Park. As a graduate student, he worked in the laboratory of Ralph J. Nossal under the graduate partnership program of the National Institutes of Health. During this time, he utilized light and neutron scattering in novel ways to shed new light on the molecular mechanisms of receptor-mediated endocytosis. At the CNRS in Montpellier, France, as an NSF postdoctoral fellow in the laboratory of Cathrine A. Royer, he implemented fluctuation microscopy to characterize gene expression and regulation in single cells of bacteria. As a postdoc at the National Cancer Institute in the laboratory of Daniel R. Larson, he pioneered in vivo single molecule studies of transcription and splicing in human cells. He is a tribal member of the Choctaw Nation of Oklahoma. He is currently an Associate Professor of Physics at Boise State University.