Confocal Microscopy: Methods and Protocols

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Stephen W. Paddock
Springer Science & Business Media, 1999 - Science - 446 pages
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In Confocal Microscopy Methods and Protocols, Stephen Paddock and a highly skilled panel of experts lead the researcher using confocal techniques from the bench top, through the imaging process, to the journal page. They concisely describe all the key stages of confocal imaging-from tissue sampling methods, through the staining process, to the manipulation, presentation, and publication of the realized image. Written in a user-friendly, nontechnical style, the methods specifically cover most of the commonly used model organisms: worms, sea urchins, flies, plants, yeast, frogs, and zebrafish. Centered in the many biological applications of the confocal microscope, the book makes possible the successful imaging of both fixed and living specimens using primarily the laser scanning confocal microscope. The powerful hands-on methods collected in Confocal Microscopy Methods and Protocols will help even the novice to produce first-class cover-quality confocal images.
 

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Contents

An Introduction to Confocal Imaging
1
Practical Considerations for Collecting Confocal Images
35
Fluorescent Probes for Confocal Microscopy
59
Imaging Gene Expression Using Antibody Probes
75
Single and Double FISH Protocols for Drosophila
93
Confocal Microscopy of Plant Cells
103
Preparation of Yeast Cells for Confocal Microscopy
131
Confocal Methods for Caenorhabditis elegans
141
Live Confocal Analysis with Fluorescently Labeled Proteins
223
Live Imaging with Green Fluorescent Protein
241
Fluorescent Calcium Indicators Subcellular Behavior and Use in Confocal Imaging
261
Intracellular pH and pCa Measurement
305
Measuring Dynamic Cell Volume In Situ by Confocal Microscopy
315
Imaging Thick Tissues with Confocal Microscopy
341
Measurement in the Confocal Microscope
357
Presentation of Confocal Images
372

Imaging Sea Urchin Fertilization
153
Imaging Immunolabeled Drosophila Embryos by Confocal Microscopy
167
Confocal Microscopy on Xenopus laevis Oocytes and Embryos
173
Analyzing Morphogenetic Cell Behaviors in Vitally Stained Zebrafish Embryos
185
Confocal Imaging of Living Cells in Intact Embryos
205
The Preparation of Stereoscopic 3D Illustrations of Confocal Data Sets for Publications and Slides
385
Information Management of Confocal Microscopy Images Traditional TextBased Databases and Image Gallery Databases
403
Morphing Confocal Images and Digital Movie Production
421
Index
443
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