Nanochannel-based single molecule recycling.

Nanochannel-based single molecule recycling.
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DOI:
10.1021/nl301341m
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发表时间:
2012-06-13
期刊:
影响因子:
10.8
通讯作者:
Novotny L
Novotny L
中科院分区:
材料科学1区
文献类型:
--
作者:
Lesoine JF;Venkataraman PA;Maloney PC;Dumont ME;Novotny L

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我们提出了一种在不固定表面的情况下测量单个分子数百次荧光的方法。这种方法是基于使用电渗透来重复驱动熔融二氧化硅纳米通道中的单个目标分子通过固定的激光焦点。在通过激光焦点的传输时间内检测到的单分子荧光被用来重复地反转控制流动方向的电势。我们的方法不依赖于连续观察,因此比现有的基于荧光的捕获方案更不容易受到荧光闪烁的影响。周转时间的变化可以用来测量单分子水平上的扩散系数。我们展示了在纳米通道中循环蛋白质和DNA的能力,并表明该过程可以与单对Förster能量转移相结合。基于纳米通道的单分子回收有望在溶液中研究同一单分子的构象动力学,而不需要表面束缚。
We present a method for measuring the fluorescence from a single molecule hundreds of times without surface immobilization. The approach is based on the use of electroosmosis to repeatedly drive a single target molecule in a fused silica nanochannel through a stationary laser focus. Single molecule fluorescence detected during the transit time through the laser focus is used to repeatedly reverse the electrical potential controlling the flow direction. Our method does not rely on continuous observation and therefore is less susceptible to fluorescence blinking than existing fluorescence-based trapping schemes. The variation in the turnaround times can be used to measure the diffusion coefficient on a single molecule level. We demonstrate the ability to recycle both proteins and DNA in nanochannels and show that the procedure can be combined with single-pair Förster energy transfer. Nanochannel-based single molecule recycling holds promise for studying conformational dynamics on the same single molecule in solution and without surface tethering.
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