通过全内反射荧光显微镜对束缚微球进行三维表征。
Three-dimensional characterization of tethered microspheres by total internal reflection fluorescence microscopy.
作者信息
Blumberg Seth, Gajraj Arivalagan, Pennington Matthew W, Meiners Jens-Christian
机构信息
Department of Physics, Biophysics Research Division, Randall Laboratory, University of Michigan, Ann Arbor, 48109-1120, USA.
出版信息
Biophys J. 2005 Aug;89(2):1272-81. doi: 10.1529/biophysj.105.061242. Epub 2005 May 27.
Abstract
Tethered particle microscopy is a powerful tool to study the dynamics of DNA molecules and DNA-protein complexes in single-molecule experiments. We demonstrate that stroboscopic total internal reflection microscopy can be used to characterize the three-dimensional spatiotemporal motion of DNA-tethered particles. By calculating characteristic measures such as symmetry and time constants of the motion, well-formed tethers can be distinguished from defective ones for which the motion is dominated by aberrant surface effects. This improves the reliability of measurements on tether dynamics. For instance, in observations of protein-mediated DNA looping, loop formation is distinguished from adsorption and other nonspecific events.
摘要
栓系粒子显微镜是在单分子实验中研究DNA分子和DNA-蛋白质复合物动力学的强大工具。我们证明频闪全内反射显微镜可用于表征DNA栓系粒子的三维时空运动。通过计算运动的对称性和时间常数等特征量，可以区分结构良好的栓系与运动受异常表面效应主导的有缺陷栓系。这提高了栓系动力学测量的可靠性。例如，在蛋白质介导的DNA环化观察中，环的形成与吸附及其他非特异性事件得以区分。
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