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.

Contents

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