In vivo fluorescence lifetime tomography of a FRET probe expressed in mouse

被引：36

作者：

McGinty, James ^{[1
]}

Stuckey, Daniel W. ^{[2
]}

Soloviev, Vadim Y. ^{[3
]}

Laine, Romain ^{[1
]}

Wylezinska-Arridge, Marzena ^{[2
]}

Wells, Dominic J. ^{[4
]}

Arridge, Simon R. ^{[3
]}

French, Paul M. W. ^{[1
]}

Hajnal, Joseph V. ^{[2
]}

Sardini, Alessandro ^{[2
]}

机构：

[1] Univ London Imperial Coll Sci Technol & Med, Blackett Lab, Photon Grp, London SW7 2BW, England

[2] Univ London Imperial Coll Sci Technol & Med, MRC Clin Sci Ctr, London W12 0NN, England

[3] UCL, Dept Comp Sci, London WC1E 6BT, England

[4] Royal Vet Coll, Dept Vet Basic Sci, London NW1 0TU, England

来源：

BIOMEDICAL OPTICS EXPRESS | 2011年 / 2卷 / 07期

基金：

英国工程与自然科学研究理事会; 英国惠康基金;

关键词：

RESONANCE ENERGY-TRANSFER; CELLS;

D O I：

10.1364/BOE.2.001907

中图分类号：

Q5 [生物化学];

学科分类号：

071010 ; 081704 ;

摘要：

Forster resonance energy transfer (FRET) is a powerful biological tool for reading out cell signaling processes. In vivo use of FRET is challenging because of the scattering properties of bulk tissue. By combining diffuse fluorescence tomography with fluorescence lifetime imaging (FLIM), implemented using wide-field time-gated detection of fluorescence excited by ultrashort laser pulses in a tomographic imaging system and applying inverse scattering algorithms, we can reconstruct the three dimensional spatial localization of fluorescence quantum efficiency and lifetime. We demonstrate in vivo spatial mapping of FRET between genetically expressed fluorescent proteins in live mice read out using FLIM. Following transfection by electroporation, mouse hind leg muscles were imaged in vivo and the emission of free donor (eGFP) in the presence of free acceptor (mCherry) could be clearly distinguished from the fluorescence of the donor when directly linked to the acceptor in a tandem (eGFP-mCherry) FRET construct. (C) 2011 Optical Society of America

引用

页码：1907 / 1917

页数：11

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