UID:
almafu_9958129773302883
Umfang:
1 online resource (878 p.)
ISBN:
9786611003579
,
9781281003577
,
1281003573
,
9780080475035
,
0080475035
Serie:
Methods in cell biology ; v. 79
Inhalt:
Recent advances in the imaging technique electron microscopy (EM) have improved the method, making it more reliable and rewarding, particularly in its description of three-dimensional detail. This book will help biologists from many disciplines understand modern EM and the value it might bring to their own work. The book's five sections deal with all major issues in EM of cells: specimen preparation, imaging in 3-D, imaging and understanding frozen-hydrated samples, labeling macromolecules, and analyzing EM data. Each chapter was written by scientists who are among the best in their field, a
Anmerkung:
Description based upon print version of record.
,
Front Cover; Methods in Cell Biology; Copyright Page; Contents; Contributors; Introduction; References; Part I: Preparing Cells for Electron Microscopy; References; Chapter 1: The Physics of Rapid Cooling and Its Implications for Cryoimmobilization of Cells; I. Introduction; II. Freezing Water; III. Biological Material; IV. Freezing Biological Material; A. Homogeneous and Heterogeneous Nucleation; B. Ice Formation in Biological Material; C. Vitrification; D. Devitrification; E. Recipe for Optimal Cryoimmobilization; Acknowledgments; References
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Chapter 2: Cryopreparation Methods for Electron Microscopy of Selected Model SystemsI. Introduction; II. Equipment and Materials; A. High-Pressure Freezers; B. Specimen Carriers; C. Accessories; D. Cryoprotectants/Fillers; E. Tools Useful for Most HPF Specimen Loading Operations; III. General Rules for Loading Samples for HPF; A. Work Only with Healthy, Unstressed Cells; B. Work Quickly; C. Do Not Let Your Cells/Tissues Dry Out; D. Avoid Surrounding Your Cells/Organisms with Aqueous Media; E. Avoid Mechanical Damage; F. Fill the Carrier Correctly; G. Use the Smallest Volume of Sample You Can
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IV. Methods for Specific OrganismsA. Yeasts (Saccharomyces cerevisiae and Schizosaccharomyces pombe); B. Nematodes (C. elegans); C. Drosophila Embryos; V. Postfreezing Processing; A. Freeze-Substitution; B. Embedding; C. FS and Embedding for Tomography; D. FS and Embedding for Immunocytochemistry; E. A.Case Study of Adjusting FS Variables to Improve Visualization; F. Sectioning; G. Microscopy and Evaluation of Results; H. Artifacts of HPF; VI. Summary; References; Chapter 3: Cryopreparation Methodology for Plant Cell Biology; I. Introduction; A. Historical Context
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B. Techniques and Organisms Discussed HereII. Rationale; A. Cryoimmobilization Versus Traditional Chemical Fixation at Ambient Temperatures: Considerations Based on First Principles; B. Why Plants and Fungi Are Usually Difficult to Fix, but Rather Easy to Freeze?; III. Methods; A. Cryoimmobilization; B. Freeze-Substitution.; C. Resin Embedding with Epoxides or LR White; D. Microtomy, Staining, and TEM; IV. Materials; V. Discussion; A. Cryoimmobilization: Choice of Freezing Method; B. Space Fillers for HPF; C. Freeze-Substitution; D. Embedding
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E. FS and Embedding Media for Microanalysis and AutoradiographyF. Other Cryopreparation Techniques; VI. Concluding Remarks; Acknowledgments; References; Chapter 4: Correlative Light and Electron Microscopy of Early Caenorhabditis elegans Embryos in Mitosis; I. Introduction; II. Rationale; III. Methods; A. Staging of Isolated Embryos by LM; B. High-Pressure Freezing and Freeze-Substitution; C. Thin-Layer Embedding, Serial Sectioning, and Prescreening of Samples; D. Electron Tomography; IV. Instrumentation and Materials; A. Staging of Early Embryos
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B. High-Pressure Freezing and Cryoprocessing of Staged Embryos
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English
Weitere Ausg.:
ISBN 9780123706478
Weitere Ausg.:
ISBN 0123706475
Sprache:
Englisch
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