Live Cell Imaging A Laboratory Manual

About this book

Recent advances in imaging technology reveal, in real time and great detail, critical changes in living cells and organisms. Live Cell Imaging is a compendium of emerging techniques, organized into two parts: specific methods such as fluorescent labeling, and delivery and detection of labeled molecules in cells; and experimental approaches ranging from the detection of single molecules to the study of dynamic processes in organelles, organs, and whole animals. Although presented primarily as a laboratory manual, Live Cell Imaging includes introductory and background material and could be used as a textbook in advanced courses. It also includes a DVD containing movies of living cells in action, created by investigators using the imaging techniques discussed in Live Cell Imaging.

Contents

Dedication
DVD Legends
Preface

Section 1: DETECTION AND APPROACHES TO LIVE CELL IMAGING
1. Fluorescent Protein Tracking and Detection M. Rizzo and D. Piston
2. Constructing and Expressing GFP Fusion Proteins D. Spector and R. Goldman
3. Viral Vectors for Introduction of GPF R. Lansford
4. Gene Delivery by Direct Injection and Facilitation of Expression by Mechanical Stretch D. Dean
5. Microinjection of Fluorophore-labeled Proteins Y. Komarova, J. Peloquin, and G. Borisy
6. CCD Cameras for Fluorescence Imaging of Living Cells P. Tran
7. Photobleaching Techniques to Study Mobility and Molecular Dynamics of Proteins in Live Cells: FRAP, iFRAP, and FLIP G. Rabut and J. Ellenberg
8. FRET and Fluorescence Lifetime Imaging Microscopy M. Tramier, D. Sanvitto, V. Emiliani, C. Durieux, and M. Coppey-Moisan
9. Monitoring Protein Dynamics Using FRET-based Biosensors T. Chew and R. Chisholm
10. Application of Light-directed Activation of Caged Biomolecules and CALI to Problems in Cell Motility D. Humphrey, Z. Rajfur, B. Imperiali, G. Marriott, P, Roy, and K. Jacobson
11. Photoactivation-based Labeling and In Vivo Tracking of RNA Molecules in the Nucleus J. Politz, R. Ruft, and T. Pederson
12. Fluorescent Speckle Microscopy of Cytoskeletal Dynamics in Living Cells T. Wittman, R. Littlefield, and C. Waterman-Storer
13. Polarization Microscopy with the LC-PolScope R. Oldenbourg
14. Confocal Microscopy, Deconvolution, and Structured Illumination Methods J. Murray
15. Multiphoton and Multispectral Laser-scanning Microscopy M. Dickinson
16. Analyzing Live Cell Data and Tracking Dynamic Movements W. Tvarusk f3;, J. Mattes, and R. Eils

SECTION 2: IMAGING OF LIVE CELLS AND ORGANISMS
17. In Vivo Imaging of Mammalian Cells J. Swedlow, P. Andrews, and M. Platani
18. A Sealed Preparation for Long-term Observation of Cultured Cells G. Sluder, J. Nordberg, F. Miller, and E. Hinchcliffe
19. Live Cell Imaging of Yeast D. Rines, D. Thomann, J. Dorn, P. Goodwin, and P. Sorger
20. Live Imaging of Caenorhabditis elegans B. Podbilewicz and Y. Gruenbaum
21. Time-lapse Cinematography in Living Drosophila Tissues I. Davis and R. Parton
22. Single-cell Imaging in Animal Tumors In Vivo J. Wyckoff, J. Segall, and J. Condeelis
23. Long-term, High-resolution Imaging in the Neocortex In Vivo B. Chen, J. Trachtenberg, A. Holtmaat, and K. Svoboda
24. Intravital Microscopy of Normal and Diseased Tissues in the Mouse R. Jain, E. Brown, L. Munn, and D. Fukumura
25. Development of Mammalian Cell Lines with lac Operator-tagged Chromosomes Y. Strukov and A. Belmont
26. Imaging Gene Expression in Living Cells S. Janicki and D. Spector
27. Studying Mitosis in Cultured Mammalian Cells P. Wadsworth
28. Imaging Hoechst-33342-labeled Chromosomes and Fluorescent Proteins During the Cell Cycle T. Haraguchi and Y. Hiraoka
29. Imaging the Actin Cytoskeleton A. Matus, V. Biou, H. Brinkhaus, and M. Roelandse
30. Imaging Intermediate Filament Proteins in Living Cells E. Kuczmarski and R. Goldman
31. Methods for Expressing and Analyzing GFP-Tubulin and GFP-Microtubule-associated Proteins H. Goodson and P. Wadsworth
32. Imaging of Organelle Membrane Systems and Membrane Traffic in Living Cells J. Lippincott-Schwartz and E. Snapp
33. Imaging Live Cells Under Mechanical Stress B. Helmke and P. Davies
34. Imaging Single Molecules Using Total Internal Reflection Fluorescence Microscopy N. Stuurman and R. Vale
35. Visualization and Quantification of Single RNA Molecules in Living Cells Y. Shav-Tal, S. Shenoy, and R. Singer

Appendix 1: Cautions
Index

Customer Reviews

Biography

The editors, David Spector and Robert Goldman, whose previous book was Cells: A Laboratory Manual, are highly respected investigators who have taught microscopy courses at Cold Spring Harbor Laboratory, the Marine Biology Laboratory at Woods Hole, and Northwestern University.