论文标题
荧光亮度，光稳定性和能量转移增强了零模式波导中固定的单分子的增强纳米剂量
Fluorescence Brightness, Photostability and Energy Transfer Enhancement of Immobilized Single Molecules in Zero-Mode Waveguides Nanoapertures
论文作者
论文摘要
零模式波导（ZMW）纳米能被广泛用于监测超出正常显微镜范围之外的单分子。但是，ZMW对荧光团光体物理学的影响的几个方面仍然没有充分记录，有时是有争议的。在这里，我们彻底研究了ZMW对单个固定的CY3B和Alexa 647分子的荧光的影响，详细介绍了亮度，寿命，光漂白时间，发射光子总数和Försterresonance Enermance Energy转移（FRET）之间的相互作用。尽管在ZMW中具有等离激元增强的激发强度，但我们发现光稳定性保留了与玻璃参考相似的光漂白时间。荧光亮度和在光漂白之前检测到的光子的总数都增加了，对于Alexa 647染料而言，发现了五倍的五倍。最后，单分子数据很重要，可以使ZMW对FRET过程的影响无漏洞的表征。我们表明，FRET速率常数增强了50％，表明纳米光子可以介导能量转移。这些结果加深了我们对ZMW中荧光增强的理解，并且与单分子生物物理应用具有直接相关性。
Zero-mode waveguide (ZMW) nanoapertures are widely used to monitor single molecules beyond the range accessible to normal microscopes. However, several aspects of the ZMW influence on the photophysics of fluorophores remain inadequately documented and sometimes controversial. Here, we thoroughly investigate the ZMW influence on the fluorescence of single immobilized Cy3B and Alexa 647 molecules, detailing the interplays between brightness, lifetime, photobleaching time, total number of emitted photons and Förster resonance energy transfer (FRET). Despite the plasmonic-enhanced excitation intensity in the ZMW, we find that the photostability is preserved with similar photobleaching times as on the glass reference. Both the fluorescence brightness and the total numbers of photons detected before photobleaching are increased, with an impressive gain near five times found for Alexa 647 dyes. Finally, the single-molecule data importantly allow a loophole-free characterization of the ZMW influence on the FRET process. We show that the FRET rate constant is enhanced by 50%, demonstrating that nanophotonics can mediate the energy transfer. These results deepen our understanding of the fluorescence enhancement in ZMWs and are of immediate relevance for single-molecule biophysical applications.
