CONFOCAL AND TWO-PHOTON MICROSCOPY FOUNDATIONS, APPLICATIONS, AND ADVANCES

Edited by

Alberto Diaspro National Institute for the Physics of Matter Department of Physics University of Genoa Genoa, Italy

®WILEY-LISS A JOHN WILEY & SONS, INC., PUBLICATION

CONTENTS

Foreword xi

G. J. Brakenhoff

Preface xüi

Acknowledgments xv

Contributors xvii

1. The Generalized Microscope 1 Colin J. R. Sheppard

2. Confocal Microscopy: Basic Principles and Architectures 19 Tony Wilson

3. Two-Photon Microscopy: Basic Principles and Architectures 39 Alberto Diaspro and Colin J. R. Sheppard

4. Cross-Sections of Fluorescent Molecules in Multiphoton Microscopy... 75 Chris Xu

5. Resolution and Contrast in Confocal and Two-Photon Microscopy . . . . 101 James E. N. Jonkman and Ernst H. K. Stelzer

6. The Role of Pinhole Size in High-Aperture Two- and Three-Photon Microscopy 127 Peter Töröok and Colin J. R. Sheppard

1. Aberrations and Penetration in In-Depth Confocal and Two-Photon-Excitation Microscopy 153 C. J. de Grauw, PL. T. M. Frederix, andH. C. Gerritsen

8. Group-Velocity Dispersion and Fiber Delivery in Multiphoton Laser Scanning Microscopy 171 Ralf Wolleschensky, Mary E. Dickinson, and Scott E. Fräser

9. Cellular and Subcellular Perturbations During Multiphoton Microscopy 191 Karsten König and Uday K. Tirlapur

VII

VÜi CONTENTS

10. Practical Multiphoton Microscopy 207 John M. Girkin and David L. Wokosin

11. Sampling, Resolution, and Digital Image Processing in the Spatial and Fourier Domains 237 Kenneth R. CasÜeman

12. Image-Restoration Methods: Basics and Algorithms 253 Patrizia Boccacci and Mario Bertero

13. Two-Photon Excitation Fluorescence Microscopy Imaging in Xenopus and Transgenic Mouse Embryos 271 Ammasi Periasamy, Cohen Nookes, Paul Skoglund, Raymond Keller, and Ann E. Sutherland

14. Multiple Color Fluorescence Imaging in Microscopy 285 Kenneth R. CasÜeman

15. Stereological Methods for Estimating Geometrical Parameters of Microscopic Structure by Three-Dimensional Imaging 299 Lucie Kubinovä, Jifi Janäcek, and Ivan Krekule

16. Imaging Live Cells in 3D Using Wide-Field Microscopy with Image Restoration 333 Walter Carrington

17. Confocal Microscopy in the Study of the Cell Nucleus 347 Luca M. Neri, Silvano Capitani, and Alberto M. Martelli

18. Confocal Imaging of Neuronal Growth and Morphology in Brain Slices 365 Irini Skaliora and Stamatis N. Pagakis

19. FISH Imaging 389 Michal Kozubek

20. Two-Photon Imaging of Tissue Physiology Based on Endogenous Fluorophores 431 Lily Hsu, Ki H. Kim, Chen Y. Dong, Peter Kaplan, Thomas Hancewicz, Christof Buehler, Keith M. Berland, Barry R. Masters, and Peter T. C So

21. Two-Photon Near-Infrared Femtosecond Laser Scanning Microscopy in Plant Biology 449 Uday K. Tirlapur and Karsten König

CONTENTS IX

22. Two-Photon Excitation Microscopy for Image Spectroscopy and Biochemistry of Tissues, Cells, Organelles, and Lipid Vesicles Under Physiological Conditions 469 Tiziana Parasassi, Luis Bagatolli, Enrico Gratton, Moshe Levi, Fluvio Ursini, Weiming Yu, and Hubert K. Zajicek

23. Real-Time In Situ Calcium Imaging with Single- and Two-Photon Confocal Microscopy 483 Katsumasa Fujita and Tetsuro Takamatsu

24. Superresolution in Fluorescence Confocal Microscopy and in DVD Optical Storage 499 Roy Pike

25. Photobleaching by Confocal Microscopy 525 James G. McNally and Carolyn L. Smith

26. Two-Photon Laser Scanning Microscopy for Characterization of Integrated Circuits and Optoelectronics 539 Chris Xu

Index 555