Journal
OPTICS LETTERS
Volume 39, Issue 20, Pages 6013-6016Publisher
OPTICAL SOC AMER
DOI: 10.1364/OL.39.006013
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Funding
- Biotechnology and Biological Sciences Research Council UK [BB/I022074/1, BB/I022937/1]
- STMicroelectronics (Imaging Division, Edinburgh, UK)
- Medical Research Council UK [MR/K015664/1]
- Dimbleby Cancer Care endowment fund
- European Community
- BBSRC [BB/I022074/1, BB/I022937/1, BB/G007160/1] Funding Source: UKRI
- MRC [MR/K015664/1] Funding Source: UKRI
- Biotechnology and Biological Sciences Research Council [BB/I022937/1, BB/I022074/1, BB/G007160/1] Funding Source: researchfish
- Medical Research Council [MR/K015664/1] Funding Source: researchfish
- Prostate Cancer UK [S12-008] Funding Source: researchfish
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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 Forster 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. (C) 2014 Optical Society of America
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