Academic Press, 2010. - 695 p. - Methods in Enzymology, Volume
475. - Single molecule tools have begun to revolutionize the molecular sciences, from biophysics to chemistry to cell biology. They hold the promise to be able to directly observe previously unseen molecular heterogeneities, quantitatively dissect complex reaction kinetics, ultimately miniaturize enzyme assays, image components of spatially distributed samples, probe the mechanical properties of single molecules in their native environment, and "just look at the thing" as anticipated by the visionary Richard Feynman already half a century ago. This volume captures a snapshot of this vibrant, rapidly expanding field, presenting articles from pioneers in the field intended to guide both the newcomer and the expert through the intricacies of getting single molecule tools.
* Includes time-tested core methods and new innovations applicable to any researcher employing single molecule tools
* Methods included are useful to both established researchers and newcomers to the field
* Relevant background and reference information given for procedures can be used as a guide to developing protocols in a number of disciplinesContents
Watching single DNA replication loops under flow extension
Star polymer surface passivation for single molecule detection
Ultrahigh resolution detection of single active motor proteins in live cells
Molecules and Methods for Superresolution Imaging
Aqueous nanodroplets for studying single molecules
High-speed atomic force microscopy techniques for visualizing dynamic behavior of biological macromolecules
Single-Biomolecule Spectroscopy Using Microfluidic Platforms
DNA Looping Kinetics Analyzed by Tethered Particle Microscopy
Single molecule observation of proteins in vivo
DNA curtains as a high-throughput approach to single molecule imaging
Single-molecule enzymology of protein synthesis
Single molecule fluorescence studies of intrinsically disordered proteins
Nanovesicle trapping for studying transient protein-protein interactions by single molecule FRET
Tracking single motor proteins in the cytoplasm of mammalian cells
Conformational States of F1-ATPase by Single-Molecule Rotation
Single Molecule Sequencing by Fluorescence Imaging
Real-Time DNA Sequencing from Single Polymerase Molecules
Micropatterning and single molecule imaging for quantitative analysis of protein-protein interactions in living cells
Probing virus-receptor interactions by atomic force spectroscopy
Single-Molecule Fluorescence Spectroscopy of Cytochrome P450 in Nanodiscs
Analysis of complex single molecule FRET time traces
Application of super-resolution imaging to single particle tracking in nanotechnology
Scanning FCS for the characterization of protein dynamics in live cells
Single mRNA molecule tracking in live cells
Single-molecule high-resolution colocalization (SHREC) or Single-molecule optical-trap analyses of protein structure
Nanopore force Spectroscopy tools for analyzing single bio-complexes
Use of plasmon coupling to reveal DNA dynamics at the single molecule level
"Fluorescence-force spectroscopy" or "Suppression of fluorophore blinking and bleaching" or "Spontaneous intersubunit rotation of the ribosome"
Multiplexed single mRNA imaging in fixed cells
Size-Minimized Quantum Dots for Single-Molecule and Intracellular Imaging
The ABEL trap
An optical torque wrench
Determining the Stoichiometry of Protein Hetero-complexes in Living Cells with Fluorescence Fluctuation Spectroscopy
Fluorescent Visualization of Single Protein-DNA Complexes
Direct Measurement of Tertiary Contact Cooperativity in RNA Folding by single molecule FRET
Nanometer-localized multiple single-molecule (NALMS) or Single-molecule nonequilibrium periodic Mg2+-concentration jump experiments
Multiparameter single molecule fluorescence detection with applications to FRET
single-particle tracking-photoactivated localization microscopy (sptPALM) within live cells
Site-specific incoporation of fluorescent probes into RNA polymerase
Quantitative single-molecule imaging by confocal laser scanning microscopy
Studies of DNA-replication at the single molecule level using magnetic tweezers
RNA labeled for single molecule FRET analysis from ligation with T4 RNA ligases
Combining optical tweezers, single-molecule fluorescence microscopy and microfluidics for studying reversible protein-DNA interactions
How dwell time distributions and other such observables in single molecule analysis can be used to extract information from molecular systems
Atomic force microscopy studies of human rhinovirus: topology and molecular forces
