Luchowski Rafal, Sabnis Sushant, Szabelski Mariusz, Sarkar Pabak, Raut Sangram, Gryczynski Zygmunt, Borejdo Julian, Bojarski Piotr, Gryczynski Ignacy
Center for Commercialization of Fluorescence Technologies (CCFT), Department of Molecular Biology & Immunology, UNTHSC, Fort Worth, TX 76107, USA.
J Lumin. 2010 Dec 1;130(12):2446-2451. doi: 10.1016/j.jlumin.2010.07.027.
Concentration is a key determining factor in the fluorescence properties of organic fluorophores. We studied self-quenching of disodium fluorescein (uranin) fluorescence in polyvinyl alcohol (PVA) thin films. The concentration dependent changes in brightness and anisotropy were followed by a lifetime decrease. We found that at a concentration of 0.54 M, the lifetime decreases to 7 ps. At a concentration of 0.18 M the lifetime was 10 ps with the relatively high quantum yield of 0.002. In these conditions the fluorescence intensity decay was homogeneous (well approximated by a single lifetime). We realized that such a sample was an ideal fluorescence lifetime standard for spectroscopy and microscopy, and therefore characterized instrument response functions for a time-domain technique. We show that self-quenched uranin enables measurements free of the color effect, making it a superior choice for a lifetime reference over scattered light.
浓度是有机荧光团荧光特性的关键决定因素。我们研究了聚乙烯醇（PVA）薄膜中荧光素钠（uranin）荧光的自猝灭现象。亮度和各向异性随浓度的变化伴随着寿命的降低。我们发现，在浓度为0.54 M时，寿命降至7 ps。在浓度为0.18 M时，寿命为10 ps，量子产率相对较高，为0.002。在这些条件下，荧光强度衰减是均匀的（由单一寿命很好地近似）。我们意识到这样的样品是光谱学和显微镜学理想的荧光寿命标准，因此对一种时域技术的仪器响应函数进行了表征。我们表明，自猝灭的uranin能够实现不受颜色效应影响的测量，使其成为比散射光更优越的寿命参考选择。
