Optical tweezers in single-molecule biophysics
Optical tweezers have become the method of choice in single-molecule manipulation studies. In this Primer, we first review the physical principles of optical tweezers and the characteristics that make them a powerful tool to investigate single molecules. We then introduce the modifications of the me...
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| Published in | Nature Reviews Methods Primers Vol. 1; no. 1; p. 25 |
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| Main Authors | , , , |
| Format | Journal Article |
| Language | English |
| Published |
England
Nature Publishing Group
25.03.2021
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| Subjects | |
| Online Access | Get full text |
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| Abstract | Optical tweezers have become the method of choice in single-molecule manipulation studies. In this Primer, we first review the physical principles of optical tweezers and the characteristics that make them a powerful tool to investigate single molecules. We then introduce the modifications of the method to extend the measurement of forces and displacements to torques and angles, and to develop optical tweezers with single-molecule fluorescence detection capabilities. We discuss force and torque calibration of these instruments, their various modes of operation and most common experimental geometries. We describe the type of data obtained in each experimental design and their analyses. This description is followed by a survey of applications of these methods to the studies of protein-nucleic acid interactions, protein/RNA folding and molecular motors. We also discuss data reproducibility, the factors that lead to the data variability among different laboratories and the need to develop field standards. We cover the current limitations of the methods and possible ways to optimize instrument operation, data extraction and analysis, before suggesting likely areas of future growth. |
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| AbstractList | Optical tweezers have become the method of choice in single-molecule manipulation studies. In this Primer, we first review the physical principles of optical tweezers and the characteristics that make them a powerful tool to investigate single molecules. We then introduce the modifications of the method to extend the measurement of forces and displacements to torques and angles, and to develop optical tweezers with single-molecule fluorescence detection capabilities. We discuss force and torque calibration of these instruments, their various modes of operation and most common experimental geometries. We describe the type of data obtained in each experimental design and their analyses. This description is followed by a survey of applications of these methods to the studies of protein-nucleic acid interactions, protein/RNA folding and molecular motors. We also discuss data reproducibility, the factors that lead to the data variability among different laboratories and the need to develop field standards. We cover the current limitations of the methods and possible ways to optimize instrument operation, data extraction and analysis, before suggesting likely areas of future growth. Optical tweezers have become the method of choice in single-molecule manipulation studies. In this Primer, we first review the physical principles of optical tweezers and the characteristics that make them a powerful tool to investigate single molecules. We then introduce the modifications of the method to extend the measurement of forces and displacements to torques and angles, and to develop optical tweezers with single-molecule fluorescence detection capabilities. We discuss force and torque calibration of these instruments, their various modes of operation and most common experimental geometries. We describe the type of data obtained in each experimental design and their analyses. This description is followed by a survey of applications of these methods to the studies of protein–nucleic acid interactions, protein/RNA folding and molecular motors. We also discuss data reproducibility, the factors that lead to the data variability among different laboratories and the need to develop field standards. We cover the current limitations of the methods and possible ways to optimize instrument operation, data extraction and analysis, before suggesting likely areas of future growth.This Primer on optical tweezers describes the instrumentation and experimental designs used in most single-molecule optical tweezers assays and discusses optical tweezers measurements in systems of biophysical interest such as DNA elasticity, protein and RNA folding, and molecular motors. |
| Author | Bustamante, Carlos J Chemla, Yann R Liu, Shixin Wang, Michelle D |
| Author_xml | – sequence: 1 givenname: Carlos J orcidid: 0000-0002-2970-0073 surname: Bustamante fullname: Bustamante, Carlos J organization: Howard Hughes Medical Institute, University of California, Berkeley, CA, USA – sequence: 2 givenname: Yann R orcidid: 0000-0001-9167-0234 surname: Chemla fullname: Chemla, Yann R organization: Department of Physics, Center for the Physics of Living Cells, University of Illinois at Urbana-Champaign, Urbana, IL, USA – sequence: 3 givenname: Shixin orcidid: 0000-0003-4238-7066 surname: Liu fullname: Liu, Shixin organization: Laboratory of Nanoscale Biophysics and Biochemistry, The Rockefeller University, New York, NY, USA – sequence: 4 givenname: Michelle D surname: Wang fullname: Wang, Michelle D organization: Department of Physics, Laboratory of Atomic and Solid State Physics, Howard Hughes Medical Institute, Cornell University, Ithaca, NY, USA |
| BackLink | https://www.ncbi.nlm.nih.gov/pubmed/34849486$$D View this record in MEDLINE/PubMed |
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| PublicationTitleAlternate | Nat Rev Methods Primers |
| PublicationYear | 2021 |
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| Snippet | Optical tweezers have become the method of choice in single-molecule manipulation studies. In this Primer, we first review the physical principles of optical... |
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| SubjectTerms | Atoms & subatomic particles Biophysics Design of experiments Electric fields Folding Force measurement Lasers Light Microscopy Molecular motors Nucleic acids Propagation Protein folding Proteins Ribonucleic acid RNA Torque |
| Title | Optical tweezers in single-molecule biophysics |
| URI | https://www.ncbi.nlm.nih.gov/pubmed/34849486 https://www.proquest.com/docview/3225862979 |
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