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Correlation of fluorescence evolution for quantitative analysis of labels and sensors
Analytica Chimica Acta ( IF 6.2 ) Pub Date : 2022-08-03 , DOI: 10.1016/j.aca.2022.340180
Agnès Pellissier-Tanon 1 , Beatrice Adelizzi 1 , Ludovic Jullien 1 , Thomas Le Saux 1 , Annie Lemarchand 2
Affiliation  

Titration without separation, e.g. quantification of a target species in living cells, is a challenge of analytical chemistry. We perform the selective detection of a target using the kinetics involved in a photochemical process and develop a correlation method that we illustrate by the titration of a fluorescent photoswitcher and the target of a photoswitching sensor. Correlating an input time series and a well-chosen weighting function associated with a variable characteristic time yields a spectrum of characteristic times. The upper integration limit of the correlation output can be chosen to match the argument of an extremum of the spectrum with a characteristic time of the input time series in order to quantify the target. A similar procedure is followed to optimize the signal-to-noise ratio. Selectivity and signal-to-noise ratio associated with 15 weighting functions are theoretically predicted. The results are applied to the titration of the reversibly photoswitchable fluorescent protein Dronpa-2 and the titration of calcium using a reversibly photoswitchable fluorescent sensor. The performance of the correlation method is favorably compared to the one of other dynamic contrast protocols.



中文翻译:

荧光演化的相关性用于标签和传感器的定量分析

没有分离的滴定,例如活细胞中目标物质的定量,是分析化学的一个挑战。我们使用光化学过程中涉及的动力学对目标进行选择性检测,并开发了一种相关方法,我们通过滴定荧光光开关和光开关传感器的目标来说明这一方法。将输入时间序列和与可变特征时间相关联的精心选择的加权函数相关联会产生特征时间的频谱。为了量化目标,可以选择相关输出的积分上限,使频谱极值的自变量与输入时间序列的特征时间相匹配。遵循类似的程序来优化信噪比。理论上预测了与 15 个加权函数相关的选择性和信噪比。将结果应用于可逆光开关荧光蛋白 Dronpa-2 的滴定和使用可逆光开关荧光传感器滴定钙。相关方法的性能优于其他动态对比协议之一。

更新日期:2022-08-03
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