定量荧光显微镜中荧光素的光漂白动力学
Photobleaching kinetics of fluorescein in quantitative fluorescence microscopy.
作者信息
Song L, Hennink E J, Young I T, Tanke H J
机构信息
Department of Cytochemistry and Cytometry, Faculty of Medicine, Leiden University, The Netherlands.
出版信息
Biophys J. 1995 Jun;68(6):2588-600. doi: 10.1016/S0006-3495(95)80442-X.
Abstract
An investigation on the photobleaching behavior of fluorescein in microscopy was carried out through a systematic analysis of photobleaching mechanisms. The individual photochemical reactions of fluorescein were incorporated into a theoretical analysis and mathematical simulation to study the photochemical processes leading to photobleaching of fluorescein in microscopy. The photobleaching behavior of free and bound fluorescein has also been investigated by experimental means. Both the theoretical simulation and experimental data show that photobleaching of fluorescein in microscopy is, in general, not a single-exponential process. The simulation suggests that the non-single-exponential behavior is caused by the oxygen-independent, proximity-induced triplet-triplet or triplet-ground state dye reactions of bound fluorescein in microscopy. The single-exponential process is a special case of photobleaching behavior when the reactions between the triplet dye and molecular oxygen are dominant.
摘要
通过对光漂白机制的系统分析，开展了荧光素在显微镜下光漂白行为的研究。将荧光素的各个光化学反应纳入理论分析和数学模拟，以研究导致显微镜下荧光素光漂白的光化学过程。还通过实验手段研究了游离和结合荧光素的光漂白行为。理论模拟和实验数据均表明，荧光素在显微镜下的光漂白通常不是单指数过程。模拟结果表明，这种非单指数行为是由显微镜下结合荧光素的与氧无关、邻近诱导的三重态-三重态或三重态-基态染料反应引起的。当三重态染料与分子氧之间的反应占主导时，单指数过程是光漂白行为的一种特殊情况。
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