Confocal and Two-Photon Microscopy Foundations, Applications, and Advances Edited by Alberto DiasproConfocal and two-photon fluorescence microscopy has provided researchers with unique possibilities of three-dimensional imaging of biological cells and tissues and of other structures such as semiconductor integrated circuits. Confocal and Two-Photon Microscopy: Foundations, Applications, and Advances provides clear, comprehensive coverage of basic foundations, modern applications, and groundbreaking new research developments made in this important area of microscopy.Opening with a foreword by G. J. Brakenhoff, this reference gathers the work of an international group of renowned experts in chapters that are logically divided into balanced sections covering theory, techniques, applications, and advances, featuring: In-depth discussion of applications for biology, medicine, physics, engineering, and chemistry, including industrial applications Guidance on new and emerging imaging technology, developmental trends, and fluorescent molecules Uniform organization and review-style presentation of chapters, with an introduction, historical overview, methodology, practical tips, applications, future directions, chapter summary, and bibliographical references Companion FTP site with full-color photographs The significant experience of pioneers, leaders, and emerging scientists in the field of confocal and two-photon excitation microscopy Confocal and Two-Photon Microscopy: Foundations, Applications, and Advances is invaluable to researchers in the biological sciences, tissue and cellular engineering, biophysics, bioengineering, physics of matter, and medicine, who use these techniques or are involved in developing new commercial instruments.

Alberto Diaspro was born in Genoa, Italy, on April 7, 1959, and went on to receive his doctoral degree in Electronic Engineering from the University of Genoa in 1983. He is an investigator and professor for the Department of Physics at the University of Genoa, and a member of the National Institute for the Physics of Matter (INFM). Dr. Diaspro also holds positions on the editorial boards of several international journals, namely: Microscopy Research and Technique, Journal of Computer Assisted Microscopy, Journal of Biomedical Optics and European Journal of Histochemistry, and he is the author of more than 100 international scientific publications. His main research activities aim at the study of biostructures both in situ and in vitro, but his many interests include the design, realization and utilization of biophysical instrumentation as conventional and confocal microscopy, two-photon fluorescence microscopy and spectroscopy architecture, differential scanning calorimetry, scanning probe microscopy (STM, SNOM, AFM), polarized light scattering, and signal and image digital processing.

Foreword

xi

G. J. Brakenhoff

Preface

xiii

Acknowledgments

xv

Contributors

xvii

The Generalized Microscope

1

(18)

Colin J. R. Sheppard

Confocal Microscopy: Basic Principles and Architectures

19

(20)

Tony Wilson

Two-Photon Microscopy: Basic Principles and Architectures

39

(36)

Alberto Diaspro

Colin J. R. Sheppard

Cross-Sections of Fluorescent Molecules in Multiphoton Microscopy

75

(26)

Chris Xu

Resolution and Contrast in Confocal and Two-Photon Microscopy

101

(26)

James E. N. Jonkman

Ernst H. K. Stelzer

The Role of Pinhole Size in High-Aperture Two-and Three-Photon Microscopy

127

(26)

Peter Torook

Colin J. R. Sheppard

Aberrations and Penetration in In-Depth Confocal and Two-Photon-Excitation Microscopy

153

(18)

C. J. de Grauw

P. L. T. M. Frederix

H. C. Gerritsen

Group-Velocity Dispersion and Fiber Delivery in Multiphoton Laser Scanning Microscopy

171

(20)

Ralf Wolleschensky

Mary E. Dickinson

Scott E. Fraser

Cellular and Subcellular Perturbations During Multiphoton Microscopy

191

(16)

Karsten Konig

Uday K. Tirlapur

Practical Multiphoton Microscopy

207

(30)

John M. Girkin

David L. Wokosin

Sampling, Resolution, and Digital Image Processing in the Spatial and Fourier Domains

237

(16)

Kenneth R. Castleman

Image-Restoration Methods: Basics and Algorithms

253

(18)

Patrizia Boccacci

Mario Bertero

Two-Photon Excitation Fluorescence Microscopy Imaging in Xenopus and Transgenic Mouse Embryos

271

(14)

Ammasi Periasamy

Colten Noakes

Paul Skoglund

Raymond Keller

Ann E. Sutherland

Multiple Color Fluorescence Imaging in Microscopy

285

(14)

Kenneth R. Castleman

Stereological Methods for Estimating Geometrical Parameters of Microscopic Structure by Three-Dimensional Imaging

299

(34)

Lucie Kubinova

Jiri Janacek

Ivan Krekule

Imaging Live Cells in 3D Using Wide-Field Microscopy with Image Restoration

333

(14)

Walter Carrington

Confocal Microscopy in the Study of the Cell Nucleus

347

(18)

Luca M. Neri

Silvano Capitani

Alberto M. Martelli

Confocal Imaging of Neuronal Growth and Morphology in Brain Slices

365

(24)

Irini Skaliora

Stamatis N. Pagakis

FISH Imaging

389

(42)

Michal Kozubek

Two-Photon Imaging of Tissue Physiology Based on Endogenous Fluorophores

431

(18)

Lily Hsu

Ki H. Kim

Chen Y. Dong

Peter Kaplan

Thomas Hancewicz

Christof Buehler

Keith M. Berland

Barry R. Masters

Peter T. C. So

Two-Photon Near-Infrared Femtosecond Laser Scanning Microscopy in Plant Biology

449

(20)

Uday K. Tirlapur

Karsten Konig

Two-Photon Excitation Microscopy for Image Spectroscopy and Biochemistry of Tissues, Cells, Organelles, and Lipid Vesicles Under Physiological Conditions

469

(14)

Tiziana Parasassi

Luis Bagatolli

Enrico Gratton

Moshe Levi

Fluvio Ursini

Weiming Yu

Hubert K. Zajicek

Real-Time In Situ Calcium Imaging with Single-and Two-Photon Confocal Microscopy

483

(16)

Katsumasa Fujita

Tetsuro Takamatsu

Superresolution in Fluorescence Confocal Microscopy and in DVD Optical Storage

499

(26)

Roy Pike

Photobleaching by Confocal Microscopy

525

(14)

James G. McNally

Carolyn L. Smith

Two-Photon Laser Scanning Microscopy for Characterization of Intergrated Circuits and Optoelectronics

539

(16)

Chris Xu

Index

555

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