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

doi:10.1364/OL.39.006013

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

^*Corresponding author for this work

Research output: Contribution to journal › Article › peer-review

36 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 language	English
Pages (from-to)	6013-6016
Number of pages	4
Journal	Optics Letters
Volume	39
Issue number	20
DOIs	https://doi.org/10.1364/OL.39.006013
Publication status	Published - 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

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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 F{\"o}rster 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.",

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AU - Poland, Simon P.

AU - Krstajić, Nikola

AU - Coelho, Simao

AU - Tyndall, David

AU - Walker, Richard J.

AU - Devauges, Viviane

AU - Morton, Penny E.

AU - Nicholas, Nicole S.

AU - Richardson, Justin

AU - Li, David Day Uei

AU - Suhling, Klaus

AU - Wells, Claire M.

AU - Parsons, Maddy

AU - Henderson, Robert K.

AU - Ameer-Beg, Simon M.

PY - 2014/10/15

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JO - Optics Letters

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

Abstract

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