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

被引：25

作者：

Hirvonen, Liisa M. ^{[1
]}

Nedbal, Jakub ^{[2
]}

Almutairi, Norah ^{[3
]}

Phillips, Thomas A. ^{[1
]}

Becker, Wolfgang ^{[4
]}

Conneely, Thomas ^{[5
]}

Milnes, James ^{[5
]}

Cox, Susan ^{[1
]}

Sturzenbaum, Stephen ^{[3
]}

Suhling, Klaus ^{[2
]}

机构：

[1] Kings Coll London, Randall Ctr Cell & Mol Biophys, London, England

[2] Kings Coll London, Dept Phys, London WC2R 2LS, England

[3] Kings Coll London, Fac Life Sci & Med, Sch Populat Hlth & Environm Sci, London, England

[4] Becker & Hickl GmbH, Berlin, Germany

[5] Photek Ltd, St Leonards On Sea, England

来源：

JOURNAL OF BIOPHOTONICS | 2020年 / 13卷 / 02期

基金：

英国工程与自然科学研究理事会; 英国生物技术与生命科学研究理事会;

关键词：

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

D O I：

10.1002/jbio.201960099

中图分类号：

Q5 [生物化学];

学科分类号：

071010 ; 081704 ;

摘要：

We report on wide-field time-correlated single photon counting (TCSPC)-based fluorescence lifetime imaging microscopy (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 microwatts with 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.

引用

页数：10

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