Würth C, Geissler D, Behnke T, Kaiser M, Resch-Genger U
BAM - Federal Institute for Materials Research and Testing, Division 1.10 Biophotonics, Richard-Willstätter-Str. 11, 12489, Berlin, Germany.
Anal Bioanal Chem. 2015 Jan;407(1):59-78. doi: 10.1007/s00216-014-8130-z. Epub 2014 Oct 23.
A crucial variable for methodical performance evaluation and comparison of luminescent reporters is the photoluminescence quantum yield (Φ pl). This quantity, defined as the number of emitted photons per number of absorbed photons, is the direct measure of the efficiency of the conversion of absorbed photons into emitted light for small organic dyes, fluorescent proteins, metal-ligand complexes, metal clusters, polymeric nanoparticles, and semiconductor and up-conversion nanocrystals. Φ pl determines the sensitivity for the detection of a specific analyte from the chromophore perspective, together with its molar-absorption coefficient at the excitation wavelength. In this review we discuss different optical and photothermal methods for measuring Φ pl of transparent and scattering systems for the most common classes of luminescent reporters, and critically evaluate their potential and limitations. In addition, reporter-specific effects and sources of uncertainty are addressed. The ultimate objective is to provide users of fluorescence techniques with validated tools for the determination of Φ pl, including a series of Φ pl standards for the ultraviolet, visible, and near-infrared regions, and to enable better judgment of the reliability of literature data.
对于发光报告分子的系统性能评估和比较而言，一个关键变量是光致发光量子产率（Φ pl）。这个量被定义为发射光子数与吸收光子数之比，它直接衡量了小分子有机染料、荧光蛋白、金属 - 配体配合物、金属簇、聚合物纳米颗粒以及半导体和上转换纳米晶体中吸收光子转化为发射光的效率。从发色团的角度来看，Φ pl与它在激发波长处的摩尔吸收系数一起，决定了检测特定分析物的灵敏度。在本综述中，我们讨论了用于测量最常见类型发光报告分子在透明和散射系统中的Φ pl的不同光学和光热方法，并批判性地评估了它们的潜力和局限性。此外，还讨论了报告分子特异性效应和不确定性来源。最终目标是为荧光技术的用户提供经过验证的用于测定Φ pl的工具，包括一系列紫外、可见和近红外区域的Φ pl标准，并使他们能够更好地判断文献数据的可靠性。
