© 2005 • 631 pp., illus., appendices, index, DVD
Paperback • $167 $83.50 • ISBN  978-087969683-2

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.

  “Overall, this new book is a superb resource for imaging techniques commonly used to improve time–lapse data, and a valuable addition to any laboratory’s library. It emphasizes the multitude of model systems that can be studied using time–lapse imaging and will surely motivate many more to join the live cell imaging revolution.”
      —Nature Cell Biology

  “The book has brought together some of the leading experts in the field of live cell imaging, each contributing a chapter that covers their particular area of expertise....Each chapter clearly introduces its subject, outlines the methods available (some with detailed protocols), and, most importantly, describes the pitfalls that can be encountered during live cell imaging. The length and detail contained is, in my opinion, ideal for gaining a good grounding in the subject of interest. I feel that this will be an excellent resource for experienced microscopists attempting live cell imaging for the first time. It would also be ideal for students wishing to learn more about the important functional information that can be gained from imaging living cells.”
      —The Quarterly Review of Biology

  “Live Cell Imaging: A Laboratory Manual, edited by Goldman and Spector, is an extremely useful addition to their previous book Cells: A Laboratory Manual.  As its name implies, it is a laboratory manual and as such contains all the pertinent information to carry out the full range of live cell imaging techniques, but it also has enough relevant background to be a useful reference manual or text for an advanced teaching course.  Not only that, it provides the necessary theory of how the various optical systems that are available work, which are most suited to what task, and in many cases provides useful practical tips from some of the best practitioners in the field on how to modify and adapt these systems to best suit individual needs.  The text is littered with examples, often including comparative studies to highlight the advantages of one technique or piece of equipment over another.  The book comes with a digital versatile disk (DVD) containing movies to further illustrate the techniques discussed.  The DVD provides extremely useful examples of what can be achieved, but also serves to highlight the realistic quality of information that can be gained by any given technique, something the naïve reader may not be fully aware of.”
      —Laboratory Animals

  “Several features of Live Cell Imaging are exemplary and can be attributed to the excellent editing of Goldman and Spector.  It is clear that the editors have extensive experience in teaching students the intricacies of Live Cell Imaging — the emphasis is on safety, caution, and compliance with university, state, and federal regulations; the book is both comprehensive and clear in its exposition; and the physical basis of each instrument or technique is described at the level of a high–quality research publication with sufficient physical and mathematical details.  There is a very good balance between the detailed description of the protocols and the validation and calibration of the resulting images and mathematical analysis.  As a much welcomed balance, the authors frequently discuss disadvantages, artifacts, and problems in an open manner: the sections on troubleshooting serve to provide the necessary solutions.  Overall, from a pedagogical viewpoint, Live Cell Imaging successfully integrates a well–written text with numerous tables of disparate data, together with full colored images, illustrations, movies on a DVD, and photographs of actual laboratory equipment.

... .The production quality of the book is very high.  The multicolor illustrations are very sharp and clearly illustrate the text.  They are complemented by easily understandable and complete figure legends.

In summary, Live Cell Imaging is well written, clear, comprehensive, well illustrated, accurate, and extremely useful as a laboratory manual or a textbook.”
      —Journal of Biomedical Optics

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