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    Adsorption and Transport of Drug-Like Molecules at the Membrane of Living Cells Studied by Time-Resolved Second-Harmonic Light Scattering

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    Genre
    Thesis/Dissertation
    Date
    2018
    Author
    Sharifian Gh., Mohammad
    Advisor
    Dai, Hai-Lung
    Committee member
    Stanley, Robert J.
    Willets, Katherine A.
    Borguet, Eric
    Yang, Weidong, Dr.
    Department
    Chemistry
    Subject
    Chemistry, Physical
    Biophysics
    Medicine
    Cell Membrane
    Membrane-specific Antimicrobial Response
    Method Development
    Nonlinear Optical Spectroscopy and Imaging of Live Cells
    Passive Membrane Permeation of Drug-like Molecules
    Second-harmonic Light Scattering
    Permanent link to this record
    http://hdl.handle.net/20.500.12613/2359
    
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    DOI
    http://dx.doi.org/10.34944/dspace/2341
    Abstract
    Understanding molecular interactions at the surfaces of cellular membranes, including adsorption and transport, is of fundamental importance in both biological and pharmaceutical studies. At present, particularly with respect to small and medium size (drug-like) molecules, it is desirable to gain an understanding of the mechanisms that govern membrane adsorption and transport. To characterize drug-membrane interactions and mechanisms governing the process of molecular uptake at cellular membranes in living organisms, we need to develop effective experimental techniques to reach quantitative and time-resolved analysis of molecules at the membrane surfaces. Also, we preferably want to develop label-free optical techniques suited for single-cell and live cell analysis. Here, I discuss the nonlinear optical technique, second-harmonic light scattering (SHS), for studying molecule-membrane interactions and transport of molecules at the membrane of living cells with real-time resolution and membrane surface-specificity. Time-resolved SHS can quantify adsorption and transport of molecules, with specific nonlinear optical properties, at living organisms without imposing any mechanical stress onto the membrane. This label-free and surface-sensitive technique can even differentiate molecular transport at individual membranes within a multi-membrane cell (e.g., bacteria). In this dissertation, I present our current research and accomplishments in extending the capabilities of the SHS technique to study molecular uptake kinetics at the membranes of living cells, to monitor bacteria membrane integrity, to characterize the antibacterial mechanism-of-action of antibiotic compounds, to update the molecular mechanism of the Gram-stain protocol, to pixel-wise mapping of the membrane viscosity of the living cells, and to probe drug-induced activation of bacterial mechanosensitive channels in vitro.
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