Malinowska Agata M, van Mameren Joost, Peterman Erwin J G, Wuite Gijs J L, Heller Iddo
LaserLaB Amsterdam and Department of Physics and Astronomy, Vrije Universiteit Amsterdam, Amsterdam, The Netherlands.
Institute of Physics, University of Amsterdam, Amsterdam, The Netherlands.
Methods Mol Biol. 2024;2694:3-28. doi: 10.1007/978-1-0716-3377-9_1.
Optical tweezers are a means to manipulate objects with light. With the technique, microscopically small objects can be held and steered, allowing for accurate measurement of the forces applied to these objects. Optical tweezers can typically obtain a nanometer spatial resolution, a picoNewton force resolution, and a millisecond time resolution, which makes the technique well suited for the study of biological processes from the single-cell down to the single-molecule level. In this chapter, we aim to provide an introduction to the use of optical tweezers for single-molecule analyses. We start from the basic principles and methodology involved in optical trapping, force calibration, and force measurements. Next, we describe the components of an optical tweezers setup and their experimental relevance. Finally, we will provide an overview of the broad applications in context of biological research, with the emphasis on the measurement modes, experimental assays, and possible combinations with fluorescence microscopy techniques.
光镊是一种用光来操纵物体的手段。通过这项技术，可以夹持和操控微观尺寸的物体，从而能够精确测量施加在这些物体上的力。光镊通常可实现纳米级的空间分辨率、皮牛级的力分辨率以及毫秒级的时间分辨率，这使得该技术非常适合用于从单细胞水平直至单分子水平的生物过程研究。在本章中，我们旨在介绍光镊在单分子分析中的应用。我们将从光阱、力校准和力测量所涉及的基本原理和方法入手。接下来，我们描述光镊装置的组成部分及其实验相关性。最后，我们将概述光镊在生物学研究背景下的广泛应用，重点介绍测量模式、实验分析方法以及与荧光显微镜技术的可能组合。
