Multivariate Curve Resolution Slicing of Multiexponential Time-Resolved Spectroscopy Fluorescence Data
Multivariate Curve Resolution Slicing of Multiexponential Time-Resolved Spectroscopy Fluorescence Data
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DOI：
10.1021/acs.analchem.1c01284
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发表时间：
2021-09-08
影响因子：
7.4
通讯作者：
Ruckebusch, Cyril
中科院分区：
文献类型：
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作者：
Devos, Olivier;Ghaffari, Mahdiyeh;Ruckebusch, Cyril
时间分辨荧光光谱(TRFs)，即测量不同激发和/或发射波长的荧光衰减曲线，提供关于分子及其环境的特定和敏感的局部信息。然而，TRFS依赖于多指数数据拟合来从测量的衰减曲线中获得荧光寿命，并且该技术的时间分辨率受到仪器响应函数(IRF)的限制。本文提出了一种基于数据切片的多元曲线分解(MCR)方法，用于对多指数荧光衰减曲线进行量身定制和无拟合度分析。MCR切片以多元曲线分解-交替最小二乘(MCR-ALS)软建模算法为基本框架，依赖于双线性/三线性混合数据分解。该方法的一个关键特征是，它能够恢复由仅部分可用单指数函数描述的衰减曲线表征的各个分量。对于TRFS数据，不仅可以分解纯多指数尾部信息，还可以分解较短的时延信息，其中由于时间分辨率的限制，信号偏离了理想的指数行为。通过分析三种商品染料的混合物并表征混合物的组成、寿命和相关贡献，验证了所提出方法的准确性，即使在只有三元混合物样品的情况下也是如此。MCR切片也被用于分析在可光切换荧光蛋白(RsEGFP2)上获得的TRFs数据。提取了三个荧光寿命以及IRF的轮廓，突出表明也可以实现复杂体系的分解，对于这些体系，单个异构体的特征是不同的指数衰减。
Time-resolved fluorescence spectroscopy (TRFS), i.e., measurement of fluorescence decay curves for different excitation and/or emission wavelengths, provides specific and sensitive local information on molecules and on their environment. However, TRFS relies on multiexponential data fitting to derive fluorescence lifetimes from the measured decay curves and the time resolution of the technique is limited by the instrumental response function (IRF). We propose here a multivariate curve resolution (MCR) approach based on data slicing to perform tailored and fit-free analysis of multiexponential fluorescence decay curves. MCR slicing, taking as a basic framework the multivariate curve resolution-alternating least-squares (MCR-ALS) soft-modeling algorithm, relies on a hybrid bilinear/trilinear data decomposition. A key feature of the method is that it enables the recovery of individual components characterized by decay profiles that are only partially describable by monoexponential functions. For TRFS data, not only pure multiexponential tail information but also shorter time delay information can be decomposed, where the signal deviates from the ideal exponential behavior due to the limited time resolution. The accuracy of the proposed approach is validated by analyzing mixtures of three commercial dyes and characterizing the mixture composition, lifetimes, and associated contributions, even in situations where only ternary mixture samples are available. MCR slicing is also applied to the analysis of TRFS data obtained on a photoswitchable fluorescent protein (rsEGFP2). Three fluorescence lifetimes are extracted, along with the profile of the IRF, highlighting that decomposition of complex systems, for which individual isomers are characterized by different exponential decays, can also be achieved.