Life scientists believe that life is driven, directed, and shaped by biomolecules working on their own or in concert. It is only in the last few decades that technological breakthroughs in sensitive fluorescence microscopy and single-molecule manipulation techniques have made it possible to observe and manipulate single biomolecules and measure their individual properties. The methodologies presented in Single Molecule Techniques: Methods and Protocols are being applied more and more to the study of biologically relevant molecules, such as DNA, DNA-binding proteins, and motor proteins, and are becoming commonplace in molecular biophysics, biochemistry, and molecular and cell biology. The aim of Single Molecule Techniques: Methods and Protocols is to provide a broad overview of single-molecule approaches applied to biomolecules on the basis of clear and concise protocols, including a solid introduction to the most widely used single-molecule techniques, such as optical tweezers, single-molecule fluorescence tools, atomic force microscopy, magnetic tweezers, and tethered particle motion. Written in the highly successful Methods in Molecular Biology™ series format, chapters contain introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and notes on troubleshooting and avoiding known pitfalls.

Authoritative and accessible, Single Molecule Techniques: Methods and Protocols serves as an ideal guide to scientists of all backgrounds and provides a broad and thorough overview of the exciting and still-emerging field of single-molecule biology.

1. Introduction to Optical Tweezers: Background, System Designs, and Commercial Solutions
Joost van Mameren, Gijs J. L. Wuite, and Iddo Heller

2. Optical Trapping and Unfolding of RNA
Katherine H. White and Koen Visscher

3. DNA Unzipping and Force Measurements with a Dual Optical Trap
Ismaïl Cissé, Pierre Mangeol, and Ulrich Bockelmann

4. Probing the Force Generation and Stepping Behavior of Cytoplasmic Dynein
Arne Gennerich and Samara L. Reck-Peterson

5. A Brief Introduction to Single-molecule Fluorescence Methods
Siet M.J.L. van den Wildenberg, Bram Prevo, and Erwin J.G. Peterman

6. Fluorescent Labeling of Proteins
Mauro Modesti

7. Fluorescence Imaging of Single Kinesin Motors on Immobilized Microtubules
Till Korten, Bert Nitzsche, Chris Gell, Felix Ruhnow, Cécile Leduc, and Stefan Diez

8. Exploring Protein Superstructures and Dynamics in Live Bacterial Cells using Single-molecule and Superresolution Imaging
Julie S. Biteen, Lucy Shapiro, and W. E. Moerner

9. Fluorescence Microscopy of Nanochannel-confined DNA
Fredrik Persson, Fredrik Westerlund, and Jonas O. Tegenfeldt

10. Fluorescence Correlation Spectroscopy
Patrick Ferrand, Jérôme Wenger, and Hervé Rigneault

11. Introduction to Atomic Force Microscopy
Pedro J. de Pablo

12. Sample Preparation for SFM Imaging of DNA, Proteins, and DNA-protein Complexes
Dejan Ristic, Humberto Sanchez, and Claire Wyman

13. Single Molecule Protein Unfolding and Refolding using Atomic Force Microscopy
Thomas Bornschlögl and Matthias Rief

14. How to Perform a Nanoindentation Experiment on a Virus
Wouter H. Roos

15. Magnetic Tweezers for Single-molecule Manipulation
Yeonee Seol and Keir C. Neuman

16. Probing DNA topology using Tethered Particle Motion
David Dunlap, Chiara Zurla, Carlo Manzo, and Laura Finzi

This book provides an overview of single-molecule approaches applied to biomolecules, including a solid introduction to the most widely used single-molecule techniques. It features readily reproducible laboratory protocols and notes on troubleshooting.

Life scientists believe that life is driven, directed, and shaped by biomolecules working on their own or in concert. It is only in the last few decades that technological breakthroughs in sensitive fluorescence microscopy and single-molecule manipulation techniques have made it possible to observe and manipulate single biomolecules and measure their individual properties. The methodologies presented in Single Molecule Techniques: Methods and Protocols are being applied more and more to the study of biologically relevant molecules, such as DNA, DNA-binding proteins, and motor proteins, and are becoming commonplace in molecular biophysics, biochemistry, and molecular and cell biology. The aim of Single Molecule Techniques: Methods and Protocols is to provide a broad overview of single-molecule approaches applied to biomolecules on the basis of clear and concise protocols, including a solid introduction to the most widely used single-molecule techniques, such as optical tweezers, single-molecule fluorescence tools, atomic force microscopy, magnetic tweezers, and tethered particle motion. Written in the highly successful Methods in Molecular Biology(TM) series format, chapters contain introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and notes on troubleshooting and avoiding known pitfalls.

Authoritative and accessible, Single Molecule Techniques: Methods and Protocols serves as an ideal guide to scientists of all backgrounds and provides a broad and thorough overview of the exciting and still-emerging field of single-molecule biology.

Introduction to Optical Tweezers: Background, System Designs, and Commercial Solutions.- Optical Trapping and Unfolding of RNA.- DNA Unzipping and Force Measurements with a Dual Optical Trap.- Probing the Force Generation and Stepping Behavior of Cytoplasmic Dynein.- A Brief Introduction to Single-molecule Fluorescence Methods.- Fluorescent Labeling of Proteins.- Fluorescence Imaging of Single Kinesin Motors on Immobilized Microtubules.- Exploring Protein Superstructures and Dynamics in Live Bacterial Cells using Single-molecule and Superresolution Imaging.- Fluorescence Microscopy of Nanochannel-confined DNA.- Fluorescence Correlation Spectroscopy.- Introduction to Atomic Force Microscopy.- Sample Preparation for SFM Imaging of DNA, Proteins, and DNA-protein Complexes.- Single Molecule Protein Unfolding and Refolding using Atomic Force Microscopy.- How to Perform a Nanoindentation Experiment on a Virus.- Magnetic Tweezers for Single-molecule Manipulation.- Probing DNA topology using Tethered Particle Motion.

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