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Multiwavelength FLIM: New applications and algorithms

机译：多波长FLIM：新应用和算法

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The combination of time-resolved and spectral resolved techniques as achieved by SLIM (spectrally resolved fluorescence lifetime imaging) improves the analysis of complex situations, when different fluorophores have to be distinguished. This could be the case when endogenous fluorophores of living cells and tissues are observed to identify the redox state and oxidative metabolic changes of the mitochondria. Other examples are FRET (resonant energy transfer) measurements, when different donor/acceptor pairs are observed simultaneously. SLIM is working in the time domain employing excitation with short light pulses and detection of the fluorescence intensity decay in many cases with time-correlated single photon counting (TCSPC). Spectral resolved detection is achieved by a polychromator in the detection path and a 16-channel multianode photomultiplier tube with the appropriate routing electronics. Within this paper special attention will be focused on FRET measurements with respect to protein interactions in Alzheimers disease. Using global analysis as the phasor plot approach or integration of the kinetic equations taking into account the multidimensional datasets in every spectral channel we could demonstrate considerable improvement of our calculations.
机译：当必须区分不同的荧光团时，通过SLIM（光谱分辨荧光寿命成像）实现的时间分辨和光谱分辨技术的组合改善了复杂情况的分析。当观察到活细胞和组织的内源荧光团以鉴定线粒体的氧化还原状态和氧化代谢变化，这可能是这种情况。当同时观察不同的供体/受体对时，其他实例是褶皱（共振能量转移）测量。在使用短的光脉冲和许多情况下使用时间相关的单光子计数（TCSPC），在使用短的光脉冲和荧光强度衰减中检测荧光强度衰减的时域。光谱分辨检测通过检测路径中的多色器和具有适当路由电子器件的16通道多年子光电倍增管实现的。在本文中，特别注意将专注于相对于阿尔茨海默病患者蛋白质疾病的蛋白质相互作用的褶皱测量。使用全局分析作为Phasor Plot方法或集成的动力学方程考虑到每个光谱通道中的多维数据集，我们可以表现出对计算的相当大的改进。

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