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Physical Chemistry

University of Colorado, Boulder

2015

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Ultrafast Nonlinear Spectroscopy Of Red Fluorescent Proteins, Patrick Eugene Konold Jan 2015

Ultrafast Nonlinear Spectroscopy Of Red Fluorescent Proteins, Patrick Eugene Konold

Chemistry & Biochemistry Graduate Theses & Dissertations (1986-2018)

Red-emitting homologues (RFPs) of the native Green Fluorescent Protein (GFP) with emission wavelengths beyond 650 nm are desirable probes for in vivo imaging experiments. They offer the potential for deeper tissue penetration and lower background scatter given a cleaner spectral window. However, bioimaging applications are hindered by poor photophysics (e.g. low fluorescence quantum yield, high photobleaching), which limits experimental resolution and represents a significant obstacle towards utilization for low copy-number, long-duration imaging applications. In this thesis, a variety of femtosecond nonlinear electronic spectroscopies were employed jointly with site-directed mutagenesis to investigate the photophysical properties of RFPs. In one study ...