High speed optically sectioned fluorescence lifetime imaging permits study of live cell signaling events

被引：68

作者：

Grant, D. M.

McGinty, J.

McGhee, E. J.

Bunney, T. D.

Owen, D. M.

Talbot, C. B.

Zhang, W.

Kumar, S.

Munro, I.

Lanigan, P. M. P.

Kennedy, G. T.

Dunsby, C.

Magee, A. I.

Courtney, P.

Katan, M.

Neil, M. A. A.

French, P. M. W.

机构：

[1] Imperial Coll London, Chem Biol Ctr, London SW7 2BW, England

[2] Imperial Coll London, Dept Phys, London SW7 2BW, England

[3] Can Res UK Ctr & Mol Biol, Chester Beatty Labs, Canc Res Inst, London SW3 6JB, England

[4] Imperial Coll London, London SW7 2AZ, England

[5] Perkin Elmer Inc, Seer Green HP9 2FX, England

来源：

OPTICS EXPRESS | 2007年 / 15卷 / 24期

基金：

英国生物技术与生命科学研究理事会; 英国医学研究理事会;

关键词：

MOLECULAR-INTERACTIONS; RAS ACTIVATION; LIVING CELLS; LIPID RAFTS; MICROSCOPY;

D O I：

10.1364/OE.15.015656

中图分类号：

O43 [光学];

学科分类号：

070207 ; 0803 ;

摘要：

We present a time domain optically sectioned fluorescence lifetime imaging (FLIM) microscope developed for high-speed live cell imaging. This single photon excited system combines wide field parallel pixel detection with confocal sectioning utilizing spinning Nipkow disc microscopy. It can acquire fluorescence lifetime images of live cells at up to 10 frames per second (fps), permitting high-speed FLIM of cell dynamics and protein interactions with potential for high throughput cell imaging and screening applications. We demonstrate the application of this FLIM microscope to real-time monitoring of changes in lipid order in cell membranes following cholesterol depletion using cyclodextrin and to the activation of the small GTP-ase Ras in live cells using FRET. (c) 2007 Optical Society of America.

引用

页码：15656 / 15673

页数：18

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