Single Molecule Tools, Part A: Fluorescence Based Approaches

  • Nils Walter

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.
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Biochemists, geneticists, molecular biologists, cell biologists, and biophysicists.


Book information

  • Published: June 2010
  • ISBN: 978-0-12-374954-3

Table of Contents

1. Watching single DNA replication loops under flow extension
2. Star polymer surface passivation for single molecule detection
3. Ultrahigh resolution detection of single active motor proteins in live cells
4. Molecules and Methods for Superresolution Imaging
5. Aqueous nanodroplets for studying single molecules
6. High-speed atomic force microscopy techniques for visualizing dynamic behavior of biological macromolecules
7. Single-Biomolecule Spectroscopy Using Microfluidic Platforms
8. DNA Looping Kinetics Analyzed by Tethered Particle Microscopy
9. Single molecule observation of proteins in vivo
10. DNA curtains as a high-throughput approach to single molecule imaging
11. Single-molecule enzymology of protein synthesis
12. Single molecule fluorescence studies of intrinsically disordered proteins
13. Nanovesicle trapping for studying transient protein-protein interactions by single molecule FRET
13. Tracking single motor proteins in the cytoplasm of mammalian cells
14. Conformational States of F1-ATPase by Single-Molecule Rotation
15. Single Molecule Sequencing by Fluorescence Imaging
16. Real-Time DNA Sequencing from Single Polymerase Molecules
17. Micropatterning and single molecule imaging for quantitative analysis of protein-protein interactions in living cells
18. Probing virus-receptor interactions by atomic force spectroscopy
19. Single-Molecule Fluorescence Spectroscopy of Cytochrome P450 in Nanodiscs
20. Analysis of complex single molecule FRET time traces
21. Application of super-resolution imaging to single particle tracking in nanotechnology
22. Scanning FCS for the characterization of protein dynamics in live cells
23. Single mRNA molecule tracking in live cells
24. Single-molecule high-resolution colocalization (SHREC) or Single-molecule optical-trap analyses of protein structure
25. Nanopore force Spectroscopy tools for analyzing single bio-complexes
26. Use of plasmon coupling to reveal DNA dynamics at the single molecule level
27. Fluorescence-force spectroscopy
28. Multiplexed single mRNA imaging in fixed cells
29. Size-Minimized Quantum Dots for Single-Molecule and Intracellular Imaging
30. The ABEL trap
31. An optical torque wrench
32. Determining the Stoichiometry of Protein Hetero-complexes in Living Cells with Fluorescence Fluctuation Spectroscopy
33. Fluorescent Visualization of Single Protein-DNA Complexes
34. Direct Measurement of Tertiary Contact Cooperativity in RNA Folding by single molecule FRET
35. Nanometer-localized multiple single-molecule (NALMS) 36. Multiparameter single molecule fluorescence detection with applications to FRET
37. Single-particle tracking-photoactivated localization microscopy (sptPALM) within live cells
38. Site-specific incoporation of fluorescent probes into RNA polymerase
39.Quantitative single-molecule imaging by confocal laser scanning microscopy
40. Studies of DNA-replication at the single molecule level using magnetic tweezers
41. RNA labeled for single molecule FRET analysis from ligation with T4 RNA ligases
42. Combining optical tweezers, single-molecule fluorescence microscopy and microfluidics for studying reversible protein-DNA interactions
43. How dwell time distributions and other such observables in single molecule analysis can be used to extract information from molecular systems