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

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

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Abstract

Imaging the spatio-temporal 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 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)6103-6106
Number of pages4
JournalOptics Letters
Volume39
Issue number20
DOIs
Publication statusPublished - 14 Oct 2014

Keywords

  • fluorescence lifetime imaging microscopy (FLIM)
  • Forster resonance energy transfer (FRET)
  • protein-protein interactions
  • multifocal multiphoton microscopy

Cite this

Poland, S. P., Krstajic, 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). A time-resolved multifocal multiphoton microscope for high speed FRET imaging in vivo. Optics Letters, 39(20), 6103-6106. https://doi.org/10.1364/OL.39.006013