Lightsheet fluorescence lifetime imaging microscopy with wide-field time-correlated single photon counting

Klaus Suhling; Liisa M. Hirvonen; Jakub Nedbal; Norah Almutairi; Thomas A. Phillips; Wolfgang Becker; Thomas Conneely; James Milnes; Susan Cox; Stephen Richard Sturzenbaum

doi:10.1002/jbio.201960099

Lightsheet fluorescence lifetime imaging microscopy with wide-field time-correlated single photon counting

Klaus Suhling, Liisa M. Hirvonen, Jakub Nedbal, Norah Almutairi, Thomas A. Phillips, Wolfgang Becker, Thomas Conneely, James Milnes, Susan Cox, Stephen Richard Sturzenbaum

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Abstract

We report onwide-field time-correlated single photon counting (TCSPC)-based fluorescence
lifetime imagingmicroscopy (FLIM) with lightsheet illumination.A pulsed
diode laser is used for excitation, and a crossed delay line anode image intensifier,
effectively a single-photon sensitive camera, is used to record the position and arrival
time of the photons with picosecond time resolution, combining low illumination
intensity of microwattswith wide-field data collection.We pair this detector with the
lightsheet illumination technique, and apply it to 3D FLIM imaging of dye gradients
in human cancer cell spheroids, and C. elegans.

Original language	English
Article number	e201960099
Journal	Journal of biophotonics
DOIs	https://doi.org/10.1002/jbio.201960099
Publication status	Accepted/In press - 2019

Keywords

SPIM
fluorescence lifetime imaging (FLIM)
lightsheet microscopy
microchannel plate (MCP)
time-correlated single photon counting (TCSPC)

UN SDGs

This output contributes to the following UN Sustainable Development Goals (SDGs)

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10.1002/jbio.201960099

191020-SPIM-FLIM-rev1Accepted author manuscript, 1.22 MB

Cite this

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title = "Lightsheet fluorescence lifetime imaging microscopy with wide-field time-correlated single photon counting",

abstract = "We report onwide-field time-correlated single photon counting (TCSPC)-based fluorescencelifetime imagingmicroscopy (FLIM) with lightsheet illumination.A pulseddiode laser is used for excitation, and a crossed delay line anode image intensifier,effectively a single-photon sensitive camera, is used to record the position and arrivaltime of the photons with picosecond time resolution, combining low illuminationintensity of microwattswith wide-field data collection.We pair this detector with thelightsheet illumination technique, and apply it to 3D FLIM imaging of dye gradientsin human cancer cell spheroids, and C. elegans.",

keywords = "SPIM, fluorescence lifetime imaging (FLIM), lightsheet microscopy, microchannel plate (MCP), time-correlated single photon counting (TCSPC)",

author = "Klaus Suhling and Hirvonen, {Liisa M.} and Jakub Nedbal and Norah Almutairi and Phillips, {Thomas A.} and Wolfgang Becker and Thomas Conneely and James Milnes and Susan Cox and Sturzenbaum, {Stephen Richard}",

year = "2019",

doi = "10.1002/jbio.201960099",

language = "English",

journal = "Journal of biophotonics",

issn = "1864-063X",

publisher = "Wiley-VCH Verlag",

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TY - JOUR

T1 - Lightsheet fluorescence lifetime imaging microscopy with wide-field time-correlated single photon counting

AU - Suhling, Klaus

AU - Hirvonen, Liisa M.

AU - Nedbal, Jakub

AU - Almutairi, Norah

AU - Phillips, Thomas A.

AU - Becker, Wolfgang

AU - Conneely, Thomas

AU - Milnes, James

AU - Cox, Susan

AU - Sturzenbaum, Stephen Richard

PY - 2019

Y1 - 2019

N2 - We report onwide-field time-correlated single photon counting (TCSPC)-based fluorescencelifetime imagingmicroscopy (FLIM) with lightsheet illumination.A pulseddiode laser is used for excitation, and a crossed delay line anode image intensifier,effectively a single-photon sensitive camera, is used to record the position and arrivaltime of the photons with picosecond time resolution, combining low illuminationintensity of microwattswith wide-field data collection.We pair this detector with thelightsheet illumination technique, and apply it to 3D FLIM imaging of dye gradientsin human cancer cell spheroids, and C. elegans.

AB - We report onwide-field time-correlated single photon counting (TCSPC)-based fluorescencelifetime imagingmicroscopy (FLIM) with lightsheet illumination.A pulseddiode laser is used for excitation, and a crossed delay line anode image intensifier,effectively a single-photon sensitive camera, is used to record the position and arrivaltime of the photons with picosecond time resolution, combining low illuminationintensity of microwattswith wide-field data collection.We pair this detector with thelightsheet illumination technique, and apply it to 3D FLIM imaging of dye gradientsin human cancer cell spheroids, and C. elegans.

KW - SPIM

KW - fluorescence lifetime imaging (FLIM)

KW - lightsheet microscopy

KW - microchannel plate (MCP)

KW - time-correlated single photon counting (TCSPC)

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U2 - 10.1002/jbio.201960099

DO - 10.1002/jbio.201960099

M3 - Article

SN - 1864-063X

JO - Journal of biophotonics

JF - Journal of biophotonics

M1 - e201960099

ER -

Lightsheet fluorescence lifetime imaging microscopy with wide-field time-correlated single photon counting

Abstract

Keywords

UN SDGs

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