用于单分子操纵与可视化的组合式磁镊与微镜全内反射荧光显微镜
Combined Magnetic Tweezers and Micro-mirror Total Internal Reflection Fluorescence Microscope for Single-Molecule Manipulation and Visualization.
作者信息
Seol Yeonee, Neuman Keir C
机构信息
Laboratory of Single Molecule Biophysics, National Heart, Lung, and Blood Institute, National Institutes of Health, Building 50, Room 3517, 50 South Drive, Bethesda, 20892, MD, USA.
出版信息
Methods Mol Biol. 2018;1665:297-316. doi: 10.1007/978-1-4939-7271-5_16.
Abstract
Magnetic tweezers is a versatile yet simple single-molecule manipulation technique that has been used to study a broad range of nucleic acids and nucleic acid-based molecular motors. In this chapter, we combine micro-mirror-based total internal reflection microscopy with a magnetic tweezers instrument, permitting simultaneous single-molecule visualization and mechanical manipulation. We provide a simple method to calibrate the evanescent wave penetration depth via supercoiling of DNA with a fluorescent nanodiamond-labeled magnetic bead and a complementary method employing a surface-immobilized fluorescent nanodiamond.
摘要
磁镊是一种通用且简单的单分子操纵技术，已被用于研究广泛的核酸和基于核酸的分子马达。在本章中，我们将基于微镜的全内反射显微镜与磁镊仪器相结合，实现了单分子的同步可视化和机械操纵。我们提供了一种简单的方法，通过用荧光纳米金刚石标记的磁珠使DNA超螺旋来校准倏逝波穿透深度，以及一种使用表面固定的荧光纳米金刚石的互补方法。
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