Construction and calibration of an optical trap on a fluorescence optical microscope
Construction and calibration of an optical trap on a fluorescence optical microscope
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DOI：
10.1038/nprot.2007.446
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发表时间：
2007-01-01
期刊：
影响因子：
14.8
通讯作者：
Dholakia, Kishan
中科院分区：
文献类型：
--
作者：
Lee, Woei Ming;Reece, Peter J.;Dholakia, Kishan
在过去的三十年里，光学陷阱的应用已经崭露头角。它们为操纵细胞、单个生物大分子、胶体微粒和纳米颗粒提供了一种强大、无菌和非侵入性的工具。光学捕获的微球可以起到力传感器的作用，用来测量皮牛顿状态下的力。通过在荧光显微镜系统中设置校准良好的单光束光学陷阱，人们可以同时测量力和收集生物系统上的荧光信号。在这个方案中，我们的目标是提供一个清晰的方法来组装和操作倒置荧光显微镜上的单光束梯度力陷阱(光镊子)。给出了调整和操作的分步指南，并讨论了常见的陷阱。
The application of optical traps has come to the fore in the last three decades. They provide a powerful, sterile and noninvasive tool for the manipulation of cells, single biological macromolecules, colloidal microparticles and nanoparticles. An optically trapped microsphere may act as a force transducer that is used to measure forces in the piconewton regime. By setting up a well-calibrated single-beam optical trap within a fluorescence microscope system, one can measure forces and collect fluorescence signals upon biological systems simultaneously. In this protocol, we aim to provide a clear exposition of the methodology of assembling and operating a single-beam gradient force trap (optical tweezers) on an inverted fluorescence microscope. A step-by-step guide is given for alignment and operation, with discussion of common pitfalls.