POLARIZED FCS

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This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. Angular fluctuations measured through polarized fluorescence correlation spectroscopy. My work has focused on exploring the various avenues for measuring angular motion using FCS. Through simulations, using SimFCS and my own programs, I have tested and evaluated the use of polarized excitation light and measuring the polarized emission. Calculating the polarization time series from the polarized dual channel emission allows one to calculate the polarization autocorrelation curve that appears to follow the angular particle rotation independent of fluctuations due to translational diffusion. This separation is not complete when one uses polarized excitation light, as we once thought. In addition to simulations, samples with slow rotations, such as dsDNA fragments, fluorophores in small unilamellar vesicles, fluorophore movement in supported bilayers, and membrane-bound proteins are being examined to evaluate the practical application of the technique.