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

Simon Poland, Nikola Krstajic, David Tyndall, Richard Walker, Justin Richardson, David Day-Uei Li, Klaus Suhling, Viviane Devauges, Simao Coelho, Penny Morton, Nicole Nicholas, Claire Wells, Maddy Parsons, Robert Henderson, Simon Ameer-Beg

Research output: Contribution to journalArticlepeer-review

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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