Szmackinski Henryk, Lakowicz Joseph R
Center for Fluorescence Spectroscopy, Department of Biological Chemistry, University of Maryland at Baltimore, School of Medicine, 108 N. Greene Street, Baltimore, MD 21201, USA
Sens Actuators B Chem. 1996 Jan 31;30(3):207-215. doi: 10.1016/0925-4005(96)80051-7.
For clinical chemistry, or for non-invasive sensing through skin, it is often necessary to obtain quantitative information in highly scattering media. We describe two simple methods for fluorescence lifetime measurements in highly scattering media, and in particular in an intralipid suspension. Lifetimes can be measured using an intensity decay law that accounts for the time delays and pulse-broadening effects of multiple light-scattering events in the intralipid. Alternatively, the phase and modulation measurements can be performed relative.to a reference fluorophore with a known lifetime. These approaches provide reliable lifetime data for a pH-sensitive fluorophore contained within a microeuvette 4 mm under the surface of the intralipid suspension. Fluorescence lifetime-based sensing is now recognized as a valuable methodology in clinical and analytical chemistry, and the possibility of lifetime-based sensing in turbid media has been demonstrated.
对于临床化学,或者对于通过皮肤进行的非侵入性传感,通常需要在高散射介质中获取定量信息。我们描述了两种在高散射介质中,特别是在脂质乳剂悬浮液中进行荧光寿命测量的简单方法。可以使用强度衰减定律来测量寿命,该定律考虑了脂质乳剂中多次光散射事件的时间延迟和脉冲展宽效应。或者,可以相对于具有已知寿命的参考荧光团进行相位和调制测量。这些方法为脂质乳剂悬浮液表面以下4毫米处的微型比色皿中所含的pH敏感荧光团提供了可靠的寿命数据。基于荧光寿命的传感现在被认为是临床和分析化学中的一种有价值的方法,并且已经证明了在混浊介质中进行基于寿命传感的可能性。