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Live Cell Imaging : A Laboratory Manual
Recent advances in imaging technology reveal, in real time and great detail, critical changes in living cells and organisms. This manual 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, the book 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 the book. 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.
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. Tvaruskó, 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