Titlebook: Single Molecule Analysis; Methods and Protocol Erwin J. G. Peterman Book 2018Latest edition Springer Science+Business Media, LLC, part of S

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Single Molecule FRET Analysis of DNA Binding Proteinsgle-molecule Förster Resonance Energy Transfer (smFRET) to probe the binding dynamics of individual proteins on single DNA molecules. We provide a detailed discussion of total internal reflection fluorescence (TIRF) instrument design, nucleic acid labeling with fluorophores, flow cell surface passivation, and data analysis methods.

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Combined Magnetic Tweezers and Micro-mirror Total Internal Reflection Fluorescence Microscope for Sint, 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.

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Introduction to Optical Tweezers: Background, System Designs, and Commercial Solutions on the trapped objects can be accurately measured and exerted. Optical tweezers can typically obtain a nanometer spatial resolution, a picoNewton force resolution, and a millisecond time resolution, which makes them excellently suited to study biological processes from the single-cell down to the s

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A Brief Introduction to Single-Molecule Fluorescence Methodsfollowing section of this volume. Fluorescence methods provide the sensitivity required to study biology on the single-molecule level, but they also allow access to useful measurable parameters on time and length scales relevant for the biomolecular world. Before several detailed experimental approa

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Fluorescent Labeling of Proteinshe movement of proteins in order to gain insight into their biological functions. A prerequisite to these experimental approaches is to graft one or more fluorophores on the protein of interest with the desired photophysical properties. Here, we describe procedures for efficient methods used to cova

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Single-Molecule Imaging of , Transmembrane Proteinse-molecule imaging due to their small size, only slightly larger than the diffraction limit of visible light. Here, we describe how reliable and reproducible single-molecule data can be obtained for a transmembrane protein in the Gram-negative bacterium . by using live-cell fluorescence microscopy.