A simple backscattering microscope for fast tracking of biological molecules
A simple backscattering microscope for fast tracking of biological molecules
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DOI：
10.1063/1.3495960
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发表时间：
2010-11-01
影响因子：
1.6
通讯作者：
Berry, Richard M.
中科院分区：
文献类型：
--
作者：
Sowa, Yoshiyuki;Steel, Bradley C.;Berry, Richard M.
在光学显微镜下观察单个分子的技术的最新发展有助于揭示分子机器的工作机制。根据研究兴趣，从单个荧光团到微米大小的标记物，可以使用广泛的标记物来检测分子。在这里，我们提出了一种新的和简单的物镜型背散射显微镜跟踪纳米和微秒分辨率的金纳米粒子。我们的系统在55 kHz带宽内的总噪声与每个轴0.6 nm相似，足以测量分子运动。我们发现我们的背散射显微镜是有用的，不仅在体外，而且在体内实验，因为较低的背景散射细胞比传统的暗场显微镜。我们证明了应用这种技术来测量的生物旋转分子马达，细菌鞭毛马达，在活大肠杆菌细胞的运动。(C)2010年美国物理学会。[doi：10.1063/1.3495960]
Recent developments in techniques for observing single molecules under light microscopes have helped reveal the mechanisms by which molecular machines work. A wide range of markers can be used to detect molecules, from single fluorophores to micron sized markers, depending on the research interest. Here, we present a new and simple objective-type backscattering microscope to track gold nanoparticles with nanometer and microsecond resolution. The total noise of our system in a 55 kHz bandwidth is similar to 0.6 nm per axis, sufficient to measure molecular movement. We found our backscattering microscopy to be useful not only for in vitro but also for in vivo experiments because of lower background scattering from cells than in conventional dark-field microscopy. We demonstrate the application of this technique to measuring the motion of a biological rotary molecular motor, the bacterial flagellar motor, in live Escherichia coli cells. (C) 2010 American Institute of Physics. [doi:10.1063/1.3495960]