Pulsed-interleaved excitation FRET measurements on single duplex DNA molecules inside C-shaped nanoapertures
Pulsed-interleaved excitation FRET measurements on single duplex DNA molecules inside C-shaped nanoapertures
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DOI：
10.1021/nl070822v
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发表时间：
2007-06-01
期刊：
影响因子：
10.8
通讯作者：
Huser, Thomas
中科院分区：
文献类型：
--
作者：
Fore, Samantha;Yuen, Yin;Huser, Thomas
单分子荧光共振能量转移（FRET）是一种被广泛接受的测定分子间空间分离的方法。结合脉冲交错激发（PIE），获得了分子相互作用化学计量学的附加信息。PIE-FRET，然而，与标准共聚焦光学实现，需要稀释样品到生物低浓度。在这里，我们展示了PIE-FRET在纳米孔径内的测量在1000倍高的浓度下产生有意义的生化数据。
Single-molecule fluorescence resonant energy transfer (FRET) is a widely accepted method for determining the spatial separation between molecules. In combination with pulsed interleaved excitation (PIE), additional information about the stoichiometry of molecular interactions is obtained. PIE-FRET, however, as implemented with standard confocal optics, requires the dilution of the sample to biologically low concentrations. Here, we show that PIE-FRET measurements inside nanometer-sized apertures yield meaningful biochemical data at 1000x higher concentrations.