Time-resolved fluorescence microscopy to study biologically related applications using sol-gel derived and cellular media

Marion Toury; Lin Chandler; Archie Allison; David Campbell; David McLoskey; A. Sheila Holmes-Smith; Graham Hungerford

doi:10.1117/12.882088

22 February 2011 Time-resolved fluorescence microscopy to study biologically related applications using sol-gel derived and cellular media

Marion Toury, Lin Chandler, Archie Allison, David Campbell, David McLoskey, A. Sheila Holmes-Smith, Graham Hungerford

Author Affiliations +

Proceedings Volume 7903, Multiphoton Microscopy in the Biomedical Sciences XI; 79031W (2011) https://doi.org/10.1117/12.882088
Event: SPIE BiOS, 2011, San Francisco, California, United States

Abstract

Fluorescence microscopy provides a non-invasive means for visualising dynamic protein interactions. As well as allowing the calculation of kinetic processes via the use of time-resolved fluorescence, localisation of the protein within cells or model systems can be monitored. These fluorescence lifetime images (FLIM) have become the preferred technique for elucidating protein dynamics due to the fact that the fluorescence lifetime is an absolute measure, in the main independent of fluorophore concentration and intensity fluctuations caused by factors such as photobleaching. In this work we demonstrate the use of a time-resolved fluorescence microscopy, employing a high repetition rate laser excitation source applied to study the influence of a metal surface on fluorescence tagged protein and to elucidate viscosity using the fluorescence lifetime probe DASPMI. These were studied in a cellular environment (yeast) and in a model system based on a sol-gel derived material, in which silver nanostructures were formed in situ using irradiation from a semiconductor laser in CW mode incorporated on a compact time-resolved fluorescence microscope (HORIBA Scientific DeltaDiode and DynaMyc).

Citation Download Citation

Marion Toury, Lin Chandler, Archie Allison, David Campbell, David McLoskey, A. Sheila Holmes-Smith, and Graham Hungerford "Time-resolved fluorescence microscopy to study biologically related applications using sol-gel derived and cellular media", Proc. SPIE 7903, Multiphoton Microscopy in the Biomedical Sciences XI, 79031W (22 February 2011); https://doi.org/10.1117/12.882088

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