© 2010 • 736 pp., illus., appendix, index
Paperback • $165 • ISBN  978-087969893-5

This title also available in: Hardcover

The movies on the accompanying website — www.cshprotocols.org/livecellimaging — display the power of recent advances in technical imaging of living cells.

The second edition of Live Cell Imaging: A Laboratory Manual expands upon and extends the collection of established and evolving methods for studying dynamic changes in living cells and organisms presented in the well-known first edition. There are 16 new chapters and the 21 updated chapters in this new edition. They include advances in atomic force microscopy, structured illumination microscopy and other 3-D approaches, as well as imaging in single cells in animals and in plants. New analytical options include live high-throughput/high-content screening in mammalian cells and computational analysis of live cell data. The manual presents hands-on techniques as well as background material, and can serve as a text in advanced courses. The first section covers principles and fundamental issues of detection and imaging; the second provides detailed protocols for imaging live systems.

SECTION 1: DETECTION AND APPROACHES TO LIVE CELL IMAGING

1. Fluorescent Protein Tracking and Detection

M.A. Rizzo, M.W. Davidson, and D.W. Piston

2. Constructing and Expressing Fluorescent Protein Fusions

D.L. Spector and R.D. Goldman

3. Micropatterning Cell-Substrate Adhesions Using Linear Polyacrylamide as the Blocking Agent

W.-h. Guo and Y.-l. Wang

4. CCD Cameras for Fluorescence Imaging of Living Cells

W.C. Salmon and J.C. Waters

5. Fluorescence Perturbation Techniques to Study Mobility and Molecular Dynamics of Proteins in Live Cells: FRAP, Photoactivation, Photoconversion, and FLIP

A. Bancaud, S. Huet, G. Rabut, and J. Ellenberg

6. Imaging Protein State in Cells

H.E. Grecco and P.I.H. Bastiaens

7. A Versatile, Multicolor Total Internal Reflection Fluorescence and Spinning-Disk Confocal Microscope System for High-Resolution Live Cell Imaging

W.D. Shin, R.S. Fischer, P. Kanchanawong, Y. Kim, J. Lim, K.A. Myers, Y. Nishimura, S.V. Plotnikov, I. Thievessen, D. Yarar, B. Sabass, and C.M. Waterman

8. Confocal Microscopy, Deconvolution, and Structured Illumination Methods

J.M. Murray

9. Atomic-Force Microscopy for Biological Imaging and Mechanical Testing across Length Scales

M. Plodinec, M. Loparic, and U. Aebi

10. OMX: A New Platform for Multimodal, Multichannel Wide-Field Imaging

I.M. Dobbie, E. King, R.M. Parton, P.M. Carlton, J.W. Sedat, J.R. Swedlow, and I. Davis

11. Digital Scanned Laser Light Sheet Fluorescence Microscopy

P.J. Keller and E.H.K. Stelzer

12. First Steps for Fluorescence Correlation Spectroscopy of Living Cells

M. Kinjo, H. Sakata, and S. Mikuni

13. Tracking and Quantitative Analysis of Dynamic Movements of Cells and Particles

K. Rohr, W.J. Godinez, N. Harder, S. Wörz, J. Mattes, W. Tvaruskó, and R. Eils

14. Imaging Techniques for Measuring the Materials Properties of Cells

K.E. Kasza, D. Vader, S. Köster, N. Wang, and D.A. Weitz

15. Computational Image Analysis of Cellular Dynamics: A Case Study Based on Particle Tracking

K. Jaqaman and G. Danuser

16. Software Tools, Data Structures, and Interfaces for Microscope Imaging

N. Stuurman and J.R. Swedlow

17. High-Throughput Microscopy Using Live Mammalian Cells

S. Terjung, T. Walter, A. Seitz, B. Neumann, R. Pepperkok, and J. Ellenberg

SECTION 2: IMAGING OF LIVE CELLS AND ORGANISMS

18. In Vivo Imaging of Mammalian Cells

J.R. Swedlow, I.M. Porter, M. Posch, and S. Swift

19. Live Cell Imaging of Yeast

D.R. Rines, D. Thomann, J.F. Dorn, P. Goodwin, and P.K. Sorger

20. Live Imaging of Caenorhabditis elegans

B. Podbilewicz and Y. Gruenbaum

21. Live Cell Imaging of Plants

Y. Fang and D.L. Spector

22. Pushing the Limits of Live Cell Imaging in Drosophila

R.M. Parton, A.M. Vallés, I.M. Dobbie, and I. Davis

23. Dynamic, Long-Term, In Vivo Imaging of Tumor-Stroma Interactions in Mouse Models of Breast Cancer Using Spinning-Disk Confocal Microscopy

A.J. Ewald, Z. Werb, and M. Egeblad

24. High-Resolution Multiphoton Imaging of Tumors In Vivo

J. Wyckoff, B. Gligorijevic, D. Entenberg, J. Segall, and J. Condeelis

25. Correlated Live Cell Light and Electron Microscopy Using Tetracysteine Tags and Biarsenicals

G.M. Gaietta, T.J. Deerinck, and M.H. Ellisman

26. Intravital Microscopy of Normal and Diseased Tissues in the Mouse

R.K. Jain, L.L. Munn, and D. Fukumura

27. Imaging Lipids in Living Cells

C. Schultz, A.B. Neef, T.W. Gadella, Jr., and J. Goedhart

28. Development of Mammalian Cell Lines with lac Operator-Tagged Chromosomes

Y.G. Strukov, M. Plutz, and A.S. Belmont

29. Imaging Gene Expression in Living Cells

S.M. Janicki and D.L. Spector

30. Studying Mitosis in Cultured Mammalian Cells

P. Wadsworth

31. Imaging Intermediate Filament Proteins in Living Cells

E.R. Kuczmarski, T. Shimi, and R.D. Goldman

32. Methods for Expressing and Analyzing GFP-Tubulin and GFP-Microtubule-Associated-Proteins

H.V. Gooodson, J.S. Dzurisin, and P. Wadsworth

33. Imaging of Membrane Systems and Membrane Traffic in Living Cells

E.L. Snapp and P. Lajoie

34. Imaging Live Cells under Mechanical Stress

B.P. Helmke and P.F. Davies

35. Imaging Single Molecules Using Total Internal Reflection Fluorescence Microscopy

S.L. Reck-Peterson, N.D.. Derr, and N. Stuurman

36. Cellular Imaging Using Total Internal Reflection Fluorescent Microscopy

D. Toomre

37. Visualization and Quantification of Single RNA Molecules in Living Cells

Y. Shav-Tal, S.M. Shenoy, and R.H. Singer

Appendix: Cautions

Index