This detailed volume explores a wide variety of techniques involving optical tweezers, a technology that has become increasingly more accessible to a broad range of researchers. Beginning with recent technical advances, the book continues by covering the application of optical tweezers to study DNA-protein interactions and DNA motors, protocols to perform protein (un)folding experiments, the application of optical tweezers to study actin- and microtubule-associated motor proteins, and well as protocols for investigating the function and mechanical properties of microtubules and intermediate filaments, and more. Written for the highly successful Methods in Molecular Biology series, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and tips on troubleshooting and avoiding known pitfalls.
Authoritative and practical, Optical Tweezers: Methodsand Protocols, Second Edition serves as an ideal resource for expanding the accessibility and use of optical traps by scientists of diverse disciplines.

Includes cutting-edge techniques Provides step-by-step detail essential for reproducible results Contains key implementation advice from the experts

Inhalt

The Invention of the Laser.- Art Ashkin and the Origins of Optical Trapping and Particle Manipulation.- Synthesis of Germanium Nanospheres as High Precision Optical Tweezers Probes.- Angular Optical Trapping to Directly Measure DNA Torsional Mechanics.- Implementation of 3D Multi-Color Fluorescence Microscopy in a Quadruple Trap Optical Tweezers System.- One-Dimensional STED Microscopy in Optical Tweezers.- Temperature Quantification and Temperature Control in Optical Tweezers.- High-Resolution Optical Tweezers Combined with Multi-Color Single-Molecule Microscopy.- Generating Negatively Supercoiled DNA Using Dual-Trap Optical Tweezers.- Force-Activated DNA Substrates for In Situ Generation of ssDNA and Designed ssDNA/dsDNA Structures in an Optical-Trapping Assay.- Probing the Interaction between Chromatin and Chromatin-Associated Complexes with Optical Tweezers.- Simultaneous Mechanical and Fluorescence Detection of Helicase-Catalyzed DNA Unwinding.- CRISPR/Cas9 On- and Off-Target Activity Using Correlative Force and Fluorescence Single-Molecule Microscopy.- Co-Temporal Single-Molecule Force and Fluorescence Measurements to Determine the Mechanism of Ribosome Translocation.- Using Single-Molecule Optical Tweezers to Study the Conformational Cycle of the Hsp90 Molecular Chaperone.- Tethering Complex Proteins and Protein Complexes for Optical Tweezers Experiments.- Single-Molecule Manipulation Study of Chaperoned SNARE Folding and Assembly with Optical Tweezers.- Using Optical Tweezers to Monitor Allosteric Signals through Changes in Folding Energy Landscapes.- High-Speed Optical Traps Address Dynamics of Processive and Non-Processive Molecular Motors.- Microtubule Dumbbells to Assess the Effect of Force Geometry on Single Kinesin Motors.- Single-Molecule Studies on the Motion and Force Generation of the Kinesin-3 Motor KIF1A.- Ultra-Fast Force-Clamp Spectroscopy of Microtubule-Binding Proteins.- Catching the Conformational Wave: Measuring the Working Strokesof Protofilaments as They Curl Outward from Disassembling Microtubule Tips.- Mechanics of Single Vimentin Intermediate Filaments under Load.- Quantifying the Interaction Strength between Biopolymers.- Measuring T Cell Receptor-Mediated Mechanosensing Using Optical Tweezers Combined with Fluorescence Imaging.