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首页> 外文期刊>The journal of physical chemistry, A. Molecules, spectroscopy, kinetics, environment, & general theory >Fluorescence Blinking and Photobleaching of Single Terrylenediimide Molecules Studied with a Confocal Microscope
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Fluorescence Blinking and Photobleaching of Single Terrylenediimide Molecules Studied with a Confocal Microscope

机译：共焦显微镜研究单Ter二酰亚胺分子的荧光闪烁和光漂白

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摘要

Single terrylenediimide molecules diluted in a 20-nm-thick polyvinylbutyral polymer film were localized and observed by scanning confocal fluorescence microscopy. A modular and compact confocal microscope and the high optical stability of the molecules allowed a repeated imaging and observation over >5 h at room temperature. Most of the molecules showed several "on-off-on" transitions (blinking) on a time scale from seconds to hours, before permanent bleaching occurred. We determined that > 1.5 * 10~7 fluorescence photons are emitted from the most-stable molecules before the final bleaching step occurs. Despite the "on-off-on" transitions, however, the overall change in fluorescence intensity, either integrated over each image of a time series or summed for several individual molecules, resembled an exponential-like decay, familiar from measurements of many-molecule ensembles. We also observed the polarization of the fluorescence from single molecules during excitation with circular polarized light. From these measurements, possible rotations of the molecular dipoles were studied. Over a span of 5 h, the polarization angle in most cases did not change by > 15-20°. This may explain the slow and small intensity changes but excludes molecular rotation as a reason for the blinking behavior.
机译：稀释在20 nm厚的聚乙烯醇缩丁醛聚合物薄膜中的单ter二酰亚胺分子被定位并通过扫描共聚焦荧光显微镜观察。模块化且紧凑的共聚焦显微镜以及分子的高光学稳定性使得在室温下> 5 h内可以重复成像和观察。在发生永久性漂白之前，大多数分子在数秒至数小时的时间内显示出数个“ on-off-on”转变（闪烁）。我们确定在最终的漂白步骤发生之前，最稳定的分子会发出> 1.5 * 10〜7的荧光光子。尽管存在“开-关-开”转换，但是荧光强度的总体变化（在时间序列的每个图像上积分或对几个单个分子求和）类似于指数型衰减，这是从多分子测量中所熟悉的合奏。我们还观察到圆偏振光激发期间单个分子的荧光的偏振。通过这些测量，研究了分子偶极子的可能旋转。在5小时的跨度内，大多数情况下的偏振角变化不大于15-20°。这可以解释强度的缓慢变化和较小变化，但不将分子旋转作为眨眼行为的原因。

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