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。
相似文献
1
Combined Magnetic Tweezers and Micro-mirror Total Internal Reflection Fluorescence Microscope for Single-Molecule Manipulation and Visualization.
Methods Mol Biol. 2018;1665:297-316. doi: 10.1007/978-1-4939-7271-5_16.
2
Correlated Single-Molecule Magnetic Tweezers and Fluorescence Measurements of DNA-Enzyme Interactions.
Methods Mol Biol. 2024;2694:421-449. doi: 10.1007/978-1-0716-3377-9_20.
3
An Introduction to Magnetic Tweezers.
Methods Mol Biol. 2024;2694:375-401. doi: 10.1007/978-1-0716-3377-9_18.
4
Simultaneous Single-Molecule Force and Fluorescence Sampling of DNA Nanostructure Conformations Using Magnetic Tweezers.
Nano Lett. 2016 Jan 13;16(1):381-6. doi: 10.1021/acs.nanolett.5b03956. Epub 2015 Dec 11.
5
High-Resolution Optical Tweezers Combined With Single-Molecule Confocal Microscopy.
Methods Enzymol. 2017;582:137-169. doi: 10.1016/bs.mie.2016.10.036. Epub 2016 Dec 14.
6
Recent Advances in Biological Single-Molecule Applications of Optical Tweezers and Fluorescence Microscopy.
Methods Enzymol. 2017;582:85-119. doi: 10.1016/bs.mie.2016.09.047. Epub 2016 Dec 12.
7
Recent advances in magnetic tweezers.
Annu Rev Biophys. 2012;41:453-72. doi: 10.1146/annurev-biophys-122311-100544. Epub 2012 Mar 14.
8
Modular magnetic tweezers for single-molecule characterizations of helicases.
Methods. 2016 Oct 1;108:4-13. doi: 10.1016/j.ymeth.2016.07.004. Epub 2016 Jul 8.
9
Magnetic tweezers: development and use in single-molecule research.
Biotechniques. 2022 Feb;72(2):65-72. doi: 10.2144/btn-2021-0104. Epub 2022 Jan 17.
10
High-Resolution Optical Tweezers Combined with Multicolor Single-Molecule Microscopy.
Methods Mol Biol. 2022;2478:141-240. doi: 10.1007/978-1-0716-2229-2_8.
引用本文的文献
1
Single-molecule DNA-flow stretching assay as a versatile hybrid tool for investigating DNA-protein interactions.
BMB Rep. 2025 Jan;58(1):41-51. doi: 10.5483/BMBRep.2024-0177.
2
Nucleosome Dynamics Derived at the Single-Molecule Level Bridges Its Structures and Functions.
JACS Au. 2024 Feb 26;4(3):866-876. doi: 10.1021/jacsau.3c00658. eCollection 2024 Mar 25.
3
General Method to Increase Carboxylic Acid Content on Nanodiamonds.
Molecules. 2022 Jan 23;27(3):736. doi: 10.3390/molecules27030736.
4
Meeting report - NSF-sponsored workshop 'Progress and Prospects of Single-Molecule Force Spectroscopy in Biological and Chemical Sciences'.
J Cell Sci. 2020 Aug 17;133(16):jcs251421. doi: 10.1242/jcs.251421.
5
Calibrating Evanescent-Wave Penetration Depths for Biological TIRF Microscopy.
Biophys J. 2019 Sep 3;117(5):795-809. doi: 10.1016/j.bpj.2019.07.048. Epub 2019 Aug 5.
本文引用的文献
1
Modular magnetic tweezers for single-molecule characterizations of helicases.
Methods. 2016 Oct 1;108:4-13. doi: 10.1016/j.ymeth.2016.07.004. Epub 2016 Jul 8.
2
Integrated magnetic tweezers and single-molecule FRET for investigating the mechanical properties of nucleic acid.
Methods. 2016 Aug 1;105:16-25. doi: 10.1016/j.ymeth.2016.06.009. Epub 2016 Jun 15.
3
Single molecule measurements of DNA helicase activity with magnetic tweezers and t-test based step-finding analysis.
Methods. 2016 Aug 1;105:119-27. doi: 10.1016/j.ymeth.2016.04.030. Epub 2016 Apr 27.
4
Simple calibration of TIR field depth using the supercoiling response of DNA.
Methods. 2016 Aug 1;105:56-61. doi: 10.1016/j.ymeth.2016.03.028. Epub 2016 Mar 30.
5
Simultaneous Single-Molecule Force and Fluorescence Sampling of DNA Nanostructure Conformations Using Magnetic Tweezers.
Nano Lett. 2016 Jan 13;16(1):381-6. doi: 10.1021/acs.nanolett.5b03956. Epub 2015 Dec 11.
6
High Spatiotemporal-Resolution Magnetic Tweezers: Calibration and Applications for DNA Dynamics.
Biophys J. 2015 Nov 17;109(10):2113-25. doi: 10.1016/j.bpj.2015.10.018.
7
Multi-wavelength single-molecule fluorescence analysis of transcription mechanisms.
Methods. 2015 Sep 15;86:27-36. doi: 10.1016/j.ymeth.2015.05.026. Epub 2015 May 30.
8
Camera-based three-dimensional real-time particle tracking at kHz rates and Ångström accuracy.
Nat Commun. 2015 Jan 7;6:5885. doi: 10.1038/ncomms6885.
9
A force calibration standard for magnetic tweezers.
Rev Sci Instrum. 2014 Dec;85(12):123114. doi: 10.1063/1.4904148.
10
Design and construction of a multiwavelength, micromirror total internal reflectance fluorescence microscope.
Nat Protoc. 2014 Oct;9(10):2317-28. doi: 10.1038/nprot.2014.155. Epub 2014 Sep 4.
Zotero 插件