XI, 381 p. 130 illus., 51 illus. in color. online resource
Confocal Microscopy: Methods and Protocols, Second Edition takes the researcher from the bench top through the imaging process, to the page. Protocols for the preparation of tissues from many model organisms including worms, flies and mice have been included as well as chapters on confocal imaging of living cells, three dimensional analysis, and the measurement and presentation of confocal images for publication. Emphasis has been placed on the laser scanning confocal microscope since this is still the instrument used for most routine applications. The current generation of modern confocal instruments produces optical sections of cells and tissues that are free of out-of-focus fluorescence with reduced chances of artifacts from the techniques of specimen preparation. This allows the imaging of living specimens and measurements of physiological events within cells. Confocal microscopy has become essential in many fields of contemporary biomedical research where a light microscope is required for imaging fluorescently labeled cells and tissues, especially cell biology, developmental biology, neurobiology, and pathology. Written in the successful Methods in Molecular Biology series format, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible protocols, and notes on troubleshooting and avoiding known pitfalls. Authoritative and easily accessible, Confocal Microscopy: Methods and Protocols, Second Edition is aimed primarily, but not exclusively, at the novice user with pointers to more advanced techniques.
Reflecting on Confocal Microscopy: A Personal Perspective.- Laser Scanning Confocal Microscopy: History, Applications And Related Optical Sectioning Techniques -- Confocal Microscopy on the Internet -- Using Photoshop with Images Created by a Confocal System -- Clearing up the Signal: Spectral Imaging and Linear Unmixing in Fluorescence Microscopy -- Low Magnification Confocal Microscopy of Tumor Angiogenesis -- Confocal Imaging of Butterfly Tissue -- Confocal Microscopy of Cardiac Myocytes -- Confocal Imaging of Fluorescently Labeled Proteins in the Drosophila Larval Neuromuscular Junction -- Confocal Imaging and Three-Dimensional Visualization of Thick Autofluorescent Specimens -- Vital Imaging of Multicellular Spheroids -- Live Confocal Analysis of Mutant and Drug Treated Drosophila Embryos -- Confocal Imaging of the Microtubule Cytoskeleton in C. elegans Embryos and Germ Cells -- Measurement in the Confocal Microscope -- A Method for Quantifying Blood Flow Distribution Among The Alveoli of the Lung -- Imaging Tools for Analysis of the Ureteric Tree in the Developing Mouse Kidney -- Evaluating Confocal Microscopy System Performance