Event DetailsEvent Dates: Monday, April 22, 2013 - 4:00pmSeminar Location: Engineering Building Room ECCR 105Speaker Name(s): Kevin DeanSpeaker Affiliation(s): University of Colorado Boulder Seminar Type/SubjectScientific Seminar Type: COSI SeminarEvent Details & Abstract: Fluorescent proteins (FPs) are powerful tools that permit real-time visualization of cellular processes. The utility of a given FP for a specific experiment depends strongly on its effective brightness, and propensity to undergo photodegradation and dark-state conversion. However, photobleaching of fluorescence is a complex phenomenon that depends on a variety of experimental and environmental factors. Under single-molecule conditions, FPs are particularly subject to photobleaching, emitting 10-100x less photons than their small - molecule counterparts. Here, we present methodology that can be used to measure irreversible photobleaching, it’s adaptation for high-throughput microfluidics-based directed evolution, and its use in the development of improved Red FPs (RFPs). Progress towards incorporation of frequency-domain fluorescence lifetime assays is also discussed. Given the quantitative and high-throughput nature of themicrofluidic photobleaching platform, this technology complements existing methods for fluorescent protein selection, thereby facilitating the development of next-generation RFPs for single-molecule research.