Time-resolved multifocal multiphoton microscope for high speed FRET imaging in vivo

Simon P. Poland, Nikola Krstajić, Simao Coelho, David Tyndall, Richard J. Walker, Viviane Devauges, Penny E. Morton, Nicole S. Nicholas, Justin Richardson, David Day Uei Li, Klaus Suhling, Claire M. Wells, Maddy Parsons, Robert K. Henderson, Simon M. Ameer-Beg

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    27 Citations (Scopus)

    Abstract

    Imaging the spatiotemporal interaction of proteins in vivo is essential to understanding the complexities of biological systems. The highest accuracy monitoring of protein-protein interactions is achieved using Förster resonance energy transfer (FRET) measured by fluorescence lifetime imaging, with measurements taking minutes to acquire a single frame, limiting their use in dynamic live cell systems. We present a diffraction limited, massively parallel, time-resolved multifocal multiphoton microscope capable of producing fluorescence lifetime images with 55 ps time-resolution, giving improvements in acquisition speed of a factor of 64. We present demonstrations with FRET imaging in a model cell system and demonstrate in vivo FLIM using a GTPase biosensor in the zebrafish embryo.

    Original languageEnglish
    Pages (from-to)6013-6016
    Number of pages4
    JournalOptics Letters
    Volume39
    Issue number20
    DOIs
    Publication statusPublished - 15 Oct 2014

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    Poland, S. P., Krstajić, N., Coelho, S., Tyndall, D., Walker, R. J., Devauges, V., Morton, P. E., Nicholas, N. S., Richardson, J., Li, D. D. U., Suhling, K., Wells, C. M., Parsons, M., Henderson, R. K., & Ameer-Beg, S. M. (2014). Time-resolved multifocal multiphoton microscope for high speed FRET imaging in vivo. Optics Letters, 39(20), 6013-6016. https://doi.org/10.1364/OL.39.006013