CAENORHABDITIS ELEGANS : Cell Biology and Physiology, 2e - Joel Rothman

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CAENORHABDITIS ELEGANS

Cell Biology and Physiology, 2e

By: Joel Rothman (Editor), Andrew Singson (Editor)

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Hardcover | 16 January 2012 | Edition Number 2

At a Glance

Hardcover
512 Pages

Edition Type
Revised

Dimensions(cm)
24.77 x 19.05 x 2.54

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The second part of an updated edition of the classic Methods in Cell Biology, Volume 48, this book emphasizes diverse methods and technologies needed to investigate C. elegans, both as an integrated organism and as a model system for research inquiries in cell, developmental, and molecular biology, as well as in genetics and pharmacology. By directing its audience to tried-and-true and cutting-edge recipes for research, this comprehensive collection is intended to guide investigators of C. elegans for years to come.

Diverse, up-to-date techniques covered will be useful to the broadening community of C. elegans researchers for years to come
Chapters written by leaders in the field
Tried and true methods deliver busy researchers a one-stop compendium of essential protocols

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Contributors	p. ix
Preface	p. xiii
Acknowledgments	p. xv
Imaging Instrumentation
High-Resolution Imaging of Cellular Processes in Caenorhabditis elegans
Microscope Stand	p. 4
Objective Lenses and Resolution	p. 5
Transmitted Light Imaging	p. 8
Fluorescence Imaging	p. 11
Detectors	p. 13
Confocal Imaging	p. 16
Deconvolution Imaging	p. 19
Whole Worm (Adult and Larval Stages)	p. 21
Embryos	p. 25
References	p. 33
Immunofluorescence Microscopy
Introduction	p. 36
Preparing Specimens for Immunolocalization Studies	p. 42
Preparation of Antibodies to Nematode Antigens	p. 54
Microscopy: Necessary Equipment and Imaging Tips	p. 58
Concluding Remarks	p. 60
Recipes	p. 60
References	p. 62
Fluorescent Protein Methods: Strategies and Applications
Introduction	p. 68
Methods	p. 70
Adopting New Fluorescent Proteins for Use in C. elegans	p. 80
References	p. 89
Modern Electron Microscopy Methods for C. elegans
Introduction	p. 94
Modern Electron Microscopes	p. 94
Alternate Immersion Fixation Techniques	p. 97
Serial Thin Sectioning	p. 116
ImmunoEM Protocols	p. 124
Alternate Methods to Post Stain Thin Sections	p. 126
Electron Tomography	p. 130
Emerging EM Technologies	p. 134
Computer-Based Analytical Tools	p. 140
Landmarks for Recognizing Tissues	p. 145
Acknowledgments	p. 146
References	p. 147
Cell Biology
Culture and Manipulation of Embryonic Cells
Introduction: Uses and Limitations of the System	p. 154
Embryo Devitellinization and Blastomere Isolation	p. 159
Culture Methods	p. 163
Drug Treatments	p. 165
Radioactive Labeling	p. 166
Fixation, Cytochemistry, and Electron Microscopy	p. 166
Solutions and Culture Media	p. 170
References	p. 174
Laser Microsurgery in Caenorhabditis elegans
Overview	p. 178
Identifying Cells in C. elegans	p. 180
Laser Ablation Theory and Apparatus	p. 186
Laser Killing of Cells	p. 192
Assessing Damage to the Operated Cell	p. 198
Unintended Damage	p. 199
Laser Cutting of Nerve Fibers	p. 199
Related Methods	p. 201
Acknowledgments	p. 203
References	p. 203
Methods in Cell Biology: Analysis of Cell Polarity in C. elegans Embryos
Introduction	p. 208
Methods for Analyzing Gene Function	p. 212
Analyzing Cell Polarity: Protein and Organelle Localization	p. 215
Dynamics of Polarity Establishment	p. 222
Summary	p. 233
References	p. 233
Analysis of Membrane-Bound Organelles
Introduction	p. 240
Visualizing Organelles	p. 240
Functional Studies	p. 250
Protein Extract and Organelle Isolation	p. 259
Summary	p. 261
Acknowledgments	p. 261
References	p. 262
C. elegans Cell Cycle Analysis
Introduction	p. 266
Types of C. elegans cell cycles	p. 266
Using the C. elegans Lineage to Follow Cell Divisions	p. 270
Methods of Developmental Synchronization	p. 272
Quantification of DNA Content	p. 275
Detection of Different Cell Cycle Phases	p. 278
Common cell Cycle Phenotypes and Potential Interpretations	p. 285
References	p. 290
-Physiology
Methods for Studying Programmed Cell Death in C. elegans
Introduction	p. 298
Methods	p. 299
Conclusions	p. 316
Acknowledgments	p. 317
References	p. 317
Methods for Studying the DNA Damage Response in the Caenorhabdatis elegans Germ Line
Introduction	p. 322
Assay Development	p. 326
Assays	p. 333
Materials	p. 347
Discussion	p. 348
Acknowledgments	p. 349
References	p. 349
Analysis of Aging in Caenorhabditis elegans
Introduction	p. 354
Aging Pathways and Their Mechanism of Action	p. 355
Life-Span Analysis	p. 358
Analysis and Interpretation of Lifespan Data	p. 367
Measurement of Age-Related Changes	p. 368
Conclusion	p. 375
Acknowledgments	p. 376
References	p. 376
Analyses of C. elegans Fat Metabolic Pathways
Introduction	p. 384
Staining of Fixed Animals and Histochemical Methods	p. 386
Biochemical Methods	p. 390
Biochemical Measurements of Metabolic Rates	p. 396
Visualization of Fat Depots and Fat Metabolism Using Vital Dyes	p. 398
Visualizing Hypodermal, Intestinal, and Gonad Lipid Stores using Nile Red	p. 400
Fatty Acid Uptake	p. 403
Raman Scattering-Based Methods of Lipid Imaging in C. elegans	p. 404
Summary	p. 405
Acknowledgements	p. 405
References	p. 406
Electrophysiological Methods for Caenorhabditis elegans Neurobiology
Introduction	p. 410
Recording Currents from Identified Neurons	p. 411
Recording Synaptic Events from Identified Neurons	p. 417
Recording Currents and Synaptic Events at the Neuromuscular Junction	p. 420
Materials	p. 428
Discussion	p. 433
Summary	p. 433
Acknowledgments	p. 434
References	p. 434
Index	p. 437
Volumes in Series	p. 449
Table of Contents provided by Ingram. All Rights Reserved.

CAENORHABDITIS ELEGANS

Cell Biology and Physiology, 2e

At a Glance

Hardcover

Shipping

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Defective items

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