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对嵌入具有背景自发荧光的散射介质中的表达增强型绿色荧光蛋白的活细胞进行荧光寿命断层扫描。

Fluorescence lifetime tomography of live cells expressing enhanced green fluorescent protein embedded in a scattering medium exhibiting background autofluorescence.

作者信息

Soloviev Vadim Y, McGinty James, Tahir Khadija B, Neil Mark A, Sardini Alessandro, Hajnal Joseph V, Arridge Simon R, French P M

机构信息

Department of Computer Science, University College London, London WC1E 6BT, UK.

出版信息

Opt Lett. 2007 Jul 15;32(14):2034-6. doi: 10.1364/ol.32.002034.

Abstract

We present a novel fluorescence lifetime tomography system applied to a highly scattering autofluorescent phantom containing live cells expressing the fluorophore enhanced green fluorescent protein (EGFP). The fluorescence signal was excited using a fiber-laser-pumped supercontinuum source and detected using wide-field time gating imaging. To facilitate rapid 3D reconstruction of the fluorescence lifetime distribution, the time-resolved data were Fourier-transformed in time to give complex functions that formed a data set for the Fourier domain reconstruction. Initially the presence of an unspecified background autofluorescence signal impeded reconstruction of the lifetime distribution, but we show that this problem can be addressed using a simple iterative technique.

摘要

我们展示了一种新型荧光寿命断层扫描系统,该系统应用于含有表达荧光团增强型绿色荧光蛋白(EGFP)的活细胞的高散射自发荧光模型。荧光信号通过光纤激光泵浦超连续光源激发,并使用宽场时间选通成像进行检测。为便于快速重建荧光寿命分布,对时间分辨数据进行时间傅里叶变换,以得到复函数,这些复函数构成用于傅里叶域重建的数据集。最初,未明确的背景自发荧光信号的存在阻碍了寿命分布的重建,但我们表明,使用一种简单的迭代技术可以解决这个问题。

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