Global analysis of fluorescence lifetime imaging microscopy data.
Global analysis of fluorescence lifetime imaging microscopy data.
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荧光寿命成像显微镜数据的全局分析。
DOI：
10.1016/s0006-3495(00)76759-2
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发表时间：
2000
影响因子：
3.4
通讯作者：
P. Bastiaens
中科院分区：
文献类型：
--
作者：
P. Verveer;A. Squire;P. Bastiaens
描述了频域荧光寿命成像显微镜（FLIM）数据的全局分析技术。这些算法利用的先验知识，只有有限数量的荧光分子物种，其寿命不变化的空间存在于样品中。描述了实现生命周期不变性约束的两种方法。在生命周期不变拟合方法中，对生命周期图像序列中的每个图像进行空间平均，以获得提高的信噪比。寿命估计从这些平均数据被用来恢复分数贡献的稳态荧光在每个物种逐像素的基础上。第二种更好的方法使用全局分析技术，同时拟合所有像素的分数贡献和空间不变的生命周期。在频域FLIM中，全局分析方法拟合的最大寿命数是传统方法拟合的最大寿命数的两倍。因此，可以用单频FLIM设置来辨别两个寿命。这些算法在模拟数据上进行了测试，然后应用于分离活细胞中共表达绿色荧光蛋白的细胞分布。
Global analysis techniques are described for frequency domain fluorescence lifetime imaging microscopy (FLIM) data. These algorithms exploit the prior knowledge that only a limited number of fluorescent molecule species whose lifetimes do not vary spatially are present in the sample. Two approaches to implementing the lifetime invariance constraint are described. In the lifetime invariant fit method, each image in the lifetime image sequence is spatially averaged to obtain an improved signal-to-noise ratio. The lifetime estimations from these averaged data are used to recover the fractional contribution to the steady-state fluorescence on a pixel-by-pixel basis for each species. The second, superior, approach uses a global analysis technique that simultaneously fits the fractional contributions in all pixels and the spatially invariant lifetimes. In frequency domain FLIM the maximum number of lifetimes that can be fit with the global analysis method is twice the number of lifetimes that can be fit with conventional approaches. As a result, it is possible to discern two lifetimes with a single-frequency FLIM setup. The algorithms were tested on simulated data and then applied to separate the cellular distributions of coexpressed green fluorescent proteins in living cells.
影响因子：
--
作者：
J. M. Beechem
通讯作者：
J. M. Beechem
使用 Fura-2 和基于寿命的传感可同时测量低钙浓度和高钙浓度。
影响因子：
4
作者：
Szmacinski,H;Lakowicz,JR
通讯作者：
Lakowicz,JR
影响因子：
4
作者：
LAKOWICZ, JR;SZMACINSKI, H;NOWACZYK, K
通讯作者：
NOWACZYK, K