Resolving single fluorophores within dense ensembles: contrast limits of tip-enhanced fluorescence microscopy
Resolving single fluorophores within dense ensembles: contrast limits of tip-enhanced fluorescence microscopy
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DOI：
10.1364/oe.16.006183
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发表时间：
2008-04-28
期刊：
影响因子：
3.8
通讯作者：
Gerton, Jordan M.
中科院分区：
文献类型：
--
作者：
Mangum, Benjamin D.;Mu, Chun;Gerton, Jordan M.
我们研究了单光子荧光作为纳米级分辨率针尖增强光学显微镜的对比机制的极限。具体来说，我们研究尖端诱导的信号增强的幅度需要解决密集包装的合奏内的个别荧光团。由振荡尖端诱导的荧光信号的调制，然后用锁定放大器解调，使图像对比度增加了近两个数量级。这个简单的调制/解调方案的理论模型预测与实验测量一致的尖端振荡幅度的最佳值。此外，作为一个重要的一步，最终应用的尖端增强荧光显微镜的纳米结构分析的生物分子系统，我们表明，必要的信号增强因子的能力范围内的市售硅提示。(c)2008年美国光学学会。
We investigate the limits of one-photon fluorescence as a contrast mechanism in nanoscale-resolution tip-enhanced optical microscopy. Specifically, we examine the magnitude of tip-induced signal enhancement needed to resolve individual fluorophores within densely-packed ensembles. Modulation of fluorescence signals induced by an oscillating tip followed by demodulation with a lock-in amplifier increases image contrast by nearly two orders of magnitude. A theoretical model of this simple modulation/demodulation scheme predicts an optimal value for the tip-oscillation amplitude that agrees with experimental measurements. Further, as an important step toward the eventual application of tip-enhanced fluorescence microscopy to the nanoscale structural analysis of biomolecular systems, we show that requisite signal enhancement factors are within the capabilities of commercially available silicon tips. (c) 2008 Optical Society of America.