Keyser U F, van der Does J, Dekker C, Dekker N H
University of Cambridge, JJ Thomson Avenue, 3 0HE, Cambridge, CB, UK.
Methods Mol Biol. 2009;544:95-112. doi: 10.1007/978-1-59745-483-4_8.
The translocation of small molecules and polymers is an integral process for the functioning of living cells. Many of the basic physical, chemical, and biological interactions have not yet been studied because they are not directly experimentally accessible. We have shown that a combination of optical tweezers, single solid-state nanopores, and electrophysiological ionic current detection enable deeper insight into the behavior of polymers in confinement. Here we describe the experimental procedures that are necessary to manipulate single biopolymers in a single nanopore, not only by electrical fields, but also through mechanical forces using optical tweezers.
小分子和聚合物的转运是活细胞功能的一个不可或缺的过程。许多基本的物理、化学和生物相互作用尚未得到研究,因为它们无法直接通过实验获取。我们已经表明,光镊、单个固态纳米孔和电生理离子电流检测相结合,能够更深入地了解聚合物在受限环境中的行为。在这里,我们描述了在单个纳米孔中操纵单个生物聚合物所需的实验程序,不仅可以通过电场,还可以使用光镊通过机械力来实现。