Handbook of Biological Confocal Microscopy

Once the second edition was safely off to the printer, the 110 larger world of micro-CT and micro-MRI and the smaller world authors breathed a sigh of relief and relaxed, secure in the belief revealed by the scanning and transmission electron microscopes. that they would "never have to do that again. " That lasted for 10 To round out the story we even have a chapter on what PowerPoint years. When we ?nally awoke, it seemed that a lot had happened. does to the results, and the annotated bibliography has been In particular, people were trying to use the Handbook as a text- updated and extended. book even though it lacked the practical chapters needed. There As with the previous editions, the editor enjoyed a tremendous had been tremendous progress in lasers and ?ber-optics and in our amount of good will and cooperation from the 124 authors understanding of the mechanisms underlying photobleaching and involved. Both I, and the light microscopy community in general, phototoxicity. It was time for a new book. I contacted "the usual owe them all a great debt of gratitude. On a more personal note, I suspects" and almost all agreed as long as the deadline was still a would like to thank Kathy Lyons and her associates at Springer for year away.

This third edition of a classic text in biological microscopy includes detailed descriptions and in-depth comparisons of parts of the microscope itself, digital aspects of data acquisition and properties of fluorescent dyes, the techniques of 3D specimen preparation and the fundamental limitations, and practical complexities of quantitative confocal fluorescence imaging. Coverage includes practical multiphoton, photodamage and phototoxicity, 3D FRET, 3D microscopy correlated with micro-MNR, CARS, second and third harmonic signals, ion imaging in 3D, scanning RAMAN, plant specimens, practical 3D microscopy and correlated optical tomography.

Foundations of Confocal Scanned Imaging in Light Microscopy.- Fundamental Limits in Confocal Microscopy.- Special Optical Elements.- Points, Pixels, and Gray Levels: Digitizing Image Data.- Laser Sources for Confocal Microscopy.- Non-Laser Light Sources for Three-Dimensional Microscopy.- Objective Lenses for Confocal Microscopy.- The Contrast Formation in Optical Microscopy.- The Intermediate Optical System of Laser-Scanning Confocal Microscopes.- Disk-Scanning Confocal Microscopy.- Measuring the Real Point Spread Function of High Numerical Aperture Microscope Objective Lenses.- Photon Detectors for Confocal Microscopy.- Structured Illumination Methods.- Visualization Systems for Multi-Dimensional Microscopy Images.- Automated Three-Dimensional Image Analysis Methods for Confocal Microscopy.- Fluorophores for Confocal Microscopy: Photophysics and Photochemistry.- Practical Considerations in the Selection and Application of Fluorescent Probes.- Guiding Principles of Specimen Preservation for Confocal Fluorescence Microscopy.- Confocal Microscopy of Living Cells.- Aberrations in Confocal and Multi-Photon Fluorescence Microscopy Induced by Refractive Index Mismatch.- Interaction of Light with Botanical Specimens.- Signal-to-Noise Ratio in Confocal Microscopes.- Comparison of Widefield/Deconvolution and Confocal Microscopy for Three-Dimensional Imaging.- Blind Deconvolution.- Image Enhancement by Deconvolution.- Fiber-Optics in Scanning Optical Microscopy.- Fluorescence Lifetime Imaging in Scanning Microscopy.- Multi-Photon Molecular Excitation in Laser-Scanning Microscopy.- Multifocal Multi-Photon Microscopy.- 4Pi Microscopy.- Nanoscale Resolution with Focused Light: Stimulated Emission Depletion and Other Reversible Saturable Optical Fluorescence Transitions Microscopy Concepts.- Mass Storage, Display, and Hard Copy.- Coherent Anti-Stokes Raman Scattering Microscopy.- Related Methods for Three-Dimensional Imaging.- Tutorial on Practical Confocal Microscopy and Use of the Confocal Test Specimen.- Practical Confocal Microscopy.- Selective Plane Illumination Microscopy.- Cell Damage During Multi-Photon Microscopy.- Photobleaching.- Nonlinear (Harmonic Generation) Optical Microscopy.- Imaging Brain Slices.- Fluorescent Ion Measurement.- Confocal and Multi-Photon Imaging of Living Embryos.- Imaging Plant Cells.- Practical Fluorescence Resonance Energy Transfer or Molecular Nanobioscopy of Living Cells.- Automated Confocal Imaging and High-Content Screening for Cytomics.- Automated Interpretation of Subcellular Location Patterns from Three-Dimensional Confocal Microscopy.- Display and Presentation Software.- When Light Microscope Resolution Is Not Enough:Correlational Light Microscopy and Electron Microscopy.- Databases for Two- and Three-Dimensional Microscopical Images in Biology.- Confocal Microscopy of Biofilms - Spatiotemporal Approaches.- Bibliography of Confocal Microscopy.