Functional in vivo imaging using fluorescence lifetime light-sheet microscopy

Claire A.  Mitchell; Simon P  Poland; James Seyforth; Jakub Nedbal; Thomas Gelot; Tahiyat Huq; Gerhard Holst; Robert D. Knight; Simon M. Ameer-Beg

doi:10.1364/OL.42.001269

Functional in vivo imaging using fluorescence lifetime light-sheet microscopy

Claire A. Mitchell, Simon P Poland, James Seyforth, Jakub Nedbal, Thomas Gelot, Tahiyat Huq, Gerhard Holst, Robert D. Knight, Simon M. Ameer-Beg^*

^*Corresponding author for this work

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

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Abstract

Light-sheet microscopy has become an indispensable tool for fast, low phototoxicity volumetric imaging of biological samples, predominantly providing structural or analyte concentration data in its standard format. Fluorescence lifetime imaging microscopy (FLIM) provides functional contrast, but often at limited acquisition speeds and with complex implementation. Therefore, we incorporate a dedicated frequency domain CMOS FLIM camera and intensity-modulated laser into a light-sheet setup to add fluorescence lifetime imaging functionality, allowing the rapid acquisition of volumetric data with concentration independent contrast. We then apply the system to image live transgenic zebrafish, demonstrating the capacity to rapidly collect volumetric FLIM data from an in vivo sample.

Original language	English
Pages (from-to)	1269-1272
Number of pages	4
Journal	Optics Letters
Volume	42
Issue number	7
DOIs	https://doi.org/10.1364/OL.42.001269
Publication status	Published - 23 Mar 2017

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Functional in vivo imaging_MITCHELL_Publishedonline27February2017_GOLD VoRFinal published version, 1.67 MBLicence: Other

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AU - Poland, Simon P

AU - Seyforth, James

AU - Nedbal, Jakub

AU - Gelot, Thomas

AU - Huq, Tahiyat

AU - Holst, Gerhard

AU - Knight, Robert D.

AU - Ameer-Beg, Simon M.

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AB - Light-sheet microscopy has become an indispensable tool for fast, low phototoxicity volumetric imaging of biological samples, predominantly providing structural or analyte concentration data in its standard format. Fluorescence lifetime imaging microscopy (FLIM) provides functional contrast, but often at limited acquisition speeds and with complex implementation. Therefore, we incorporate a dedicated frequency domain CMOS FLIM camera and intensity-modulated laser into a light-sheet setup to add fluorescence lifetime imaging functionality, allowing the rapid acquisition of volumetric data with concentration independent contrast. We then apply the system to image live transgenic zebrafish, demonstrating the capacity to rapidly collect volumetric FLIM data from an in vivo sample.

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Functional in vivo imaging using fluorescence lifetime light-sheet microscopy

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