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Single-Molecule Techniques: A Laboratory Manual

Subject Area(s):  Cell BiologyMolecular BiologyBiochemistryBiophysicsLaboratory Techniques

Edited by Paul R. Selvin, University of Illinois, Urbana-Champaign; Taekjip Ha, University of Illinois, Urbana-Champaign

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© 2008 • 507 pp., illus., appendix, index
Paperback •
ISBN  978-087969775-4

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As molecular and cellular biologists move toward nano–techniques for performing experiments on single molecules rather than on populations of molecules, a comprehensive manual on how (and why) to carry out such experiments is needed. Single-Molecule Techniques: A Laboratory Manual fills this requirement—it is the first to take researchers who know nothing about single–molecule analyses to the point where they can successfully design and execute appropriate experiments. Geared toward research scientists in structural and molecular biology, biochemistry, and biophysics, the manual will be useful to all who are interested in observing, manipulating, and elucidating the molecular mechanisms and discrete properties of macromolecules. Techniques range from in vivo and in vitro fluorescent–based methods to the use of atomic force microscopy, optical and magnetic tweezers, and nanopores. The book is edited by Paul R. Selvin and Taekjip Ha, two pioneers in the field of experimental biophysics who have made significant contributions to the development and application of single–molecule technologies.


1. The New Era of Biology In Singulo
T. Ha and P.R. Selvin

2. Single-Molecule FRET with Total Internal Reflection Microscopy
C. Joo and T. Ha
3. In Vitro and In Vivo FIONA and Other Acronyms for Watching Molecular Motors Walk
P.R. Selvin, T. Lougheed, M.T. Hoffman, H. Park, H. Balci, B.H. Blehm, and E. Toprak
4. Colocalization of Fluorescent Probes: Accurate and Precise Registration with Nanometer Resolution
L.S. Churchman and J.A. Spudich
5. Alternating-Laser Excitation of Single Molecules
A.N. Kapanidis, M. Heilemann, E. Margeat, X. Kong, E. Nir, and S. Weiss
6. Orientation and Rotational Motions of Single Molecules by Polarized Total Internal Reflection Fluorescence Microscopy
J.F. Beausang, Y. Sun, M.E. Quinlan, J.N. Forkey, and Y.E. Goldman

7. Imaging Gene Expression in Living Cells at the Single-Molecule Level
J. Xiao, J. Elf, G.-W. Li, J. Yu, and X.S. Xie
8. Single-Virus Tracking in Live Cells
M.J. Rust, M. Lakadamyali, B. Brandenburg, and X. Zhuang
9. Ultrasensitive Imaging in Live Cells Using Fluorescent Quantum Dots
S. Courty and M. Dahan
10. Imaging Real-Time Gene Expression in Living Systems
A.L. Wells, J.S. Condeelis, R.H. Singer, and D. Zenklusen
11. Single-Molecule Imaging of Stochastic Signaling Events in Living Cells
S. Matsuoka, Y. Miyanaga, T. Yanagida, and M. Ueda
12. Fluorescence Correlation Spectroscopy In Vitro and In Vivo
E. Haustein and P. Schwille

13. Optical Traps to Study Properties of Molecular Motors
J.A. Spudich, S.E. Rice, R.S. Rock, T.J. Purcell, and H.M. Warrick
14. High-Resolution Dual-Trap Optical Tweezers with Differential Detection
C. Bustamante, Y.R. Chemla, and J.R. Moffitt
15. Imaging and Nanomanipulation of an Actomyosin Motor
S. Nishikawa, T. Komori, T. Ariga, T. Okada, M. Morimatsu, Y. Ishii, and T. Yanagida

16. Single-Molecule Studies Using Magnetic Traps
T. Lionnet, J.-F. Allemand, A. Revyakin, T.R. Strick, O.A. Saleh, D. Bensimon, and V. Croquette

17. Probing Polysaccharide and Protein Mechanics by Atomic Force Microscopy
M. Rabbi and P.E. Marszalek

18. Single-Molecule Analysis of Nucleic Acids and DNA–Protein Interactions Using Nanopores
M. Wanunu and A. Meller

19. Single-Molecule Gold-Nanoparticle Tracking with High Temporal and Spatial Resolution and without an Applied Load
A.R. Dunn and J. A. Spudich
20. Advances in Surface-based Assays for Single Molecules
P.M. Fordyce, M.T. Valentine, and S.M. Block
21. Hydrodynamic Flow-stretching Assay for Single-Molecule Studies of Nucleic Acid–Protein Interactions
C.M. Schroeder, P.C. Blainey, S. Kim, and X.S. Xie

APPENDIX: Cautions



review:  “I don't believe I have ever before read a microscopy textbook from cover to cover — but this one, reader, I did. Its two editors, Taekjip Ha and Paul Selvin, have set out to assemble a manual for cell biologists who want to extend their technical repertoires to include single-molecule techniques. They have succeeded admirably, and as a bonus the book provides a fascinating insight into where the technical limits lie, of how we stand — as the authors suggest in their introduction — at the beginning of a new era of single-molecule-based cell biology...This excellent book will be invaluable to anyone who is already competent at multimolecule data collection and interested in getting down to molecular mechanisms.”
      —Nature Cell Biology