BAM Federal Institute for Materials Research and Testing, Berlin, Germany.
Anal Chem. 2011 May 1;83(9):3431-9. doi: 10.1021/ac2000303. Epub 2011 Apr 7.
The photoluminescence quantum yield (Φ(f)) that presents a direct measure for the efficiency of the conversion of absorbed photons into emitted photons is one of the spectroscopic key parameters of functional fluorophores. It determines the suitability of such materials for applications in, for example, (bio)analysis, biosensing, and fluorescence imaging as well as as active components in optical devices. The reborn interest in accurate Φ(f) measurements in conjunction with the controversial reliability of reported Φ(f) values of many common organic dyes encouraged us to compare two relative and one absolute fluorometric method for the determination of the fluorescence quantum yields of quinine sulfate dihydrate, coumarin 153, fluorescein, rhodamine 6G, and rhodamine 101. The relative methods include the use of a chain of Φ(f) transfer standards consisting of several "standard dye" versus "reference dye" pairs linked to a golden Φ(f) standard that covers the ultraviolet and visible spectral region, and the use of different excitation wavelengths for standard and sample, respectively. Based upon these measurements and the calibration of the instruments employed, complete uncertainty budgets for the resulting Φ(f) values are derived for each method, thereby providing evaluated standard operation procedures for Φ(f) measurements and, simultaneously, a set of assessed Φ(f) standards.
荧光量子产率（Φ(f)）是衡量吸收光子转化为发射光子效率的直接指标，是功能荧光团的光谱关键参数之一。它决定了这些材料在（生物）分析、生物传感和荧光成像以及光学器件中的有源组件等应用中的适用性。人们对准确测量Φ(f)的兴趣重新燃起，再加上许多常见有机染料的报告Φ(f)值的可靠性存在争议，这促使我们比较了两种相对法和一种绝对法，以测定硫酸奎宁二水合物、香豆素 153、荧光素、若丹明 6G 和若丹明 101 的荧光量子产率。相对法包括使用一系列由几个“标准染料”对“参考染料”组成的Φ(f)转移标准，这些标准与覆盖紫外可见光谱区域的金Φ(f)标准相连接，以及分别为标准和样品使用不同的激发波长。基于这些测量值和所使用仪器的校准，为每种方法推导了完整的不确定度预算，从而为Φ(f)测量提供了经过评估的标准操作程序，同时还提供了一组经过评估的Φ(f)标准。
