Introduction to Electron Microscopy for Biologists : Methods in Cell Biology
This volume demonstrates how cellular and associated electron microscopy contributes to knowledge about biological structural information, primarily at the nanometer level. It presents how EM approaches complement both conventional structural biology (at the high end, angstrom level of resolution) and digital light microscopy (at the low end, 100-200 nanometers). *Basic techniques in transmission and scanning electron microscopy *Detailed chapters on how to use electron microscopy when dealing with specific cellular structures, such as the nucleus, cell membrane, and cytoskeleto
Methods in cell biology, v. 88
1 online resource (562 pages).
9780080888163, 008088816X
1058470290
Cover; Copyright Page; TOCContents; Contributors; Part I: Exploring the Organisation of the Cell by Electron Microscopy; Section 1: Basic Transmission and Scanning Electron Microscopy; CHChapter 1: High Pressure Freezing and Freeze Substitution of Schizosaccharomyces pombe and Saccharomyces cerevisiae for TEM; I. Introduction; II. Materials and Instrumentation; III. Procedures; IV. Comments and Problems; References; CHChapter 2: Electron Probe X-ray Microanalysis for the Study of Cell Physiology; I. Introduction; II. Rationale; III. Methods; IV. Equipment; V. Discussion; References. CHChapter 3: Preparation of Cells and Tissues for Immuno EMI. Preparing Biological Specimens for Examination by Electron Microscopy; II. Vitrification and Chemical Fixation for Immunolocalization; III. Vitrification Followed by Freeze Substitution; IV. Chemical Cross-linking (or Fixation); V. Embedding in Resin for Sectioning; VI. Cryosectioning for Immunocytochemistry: The Tokuyasu Method; VII. The Starting Material; VIII. Protocols; IX. Alternative Approaches to Freeze Substitution; X. Correlative Microscopy; References. CHChapter 4: Quantification of Structures and Gold Labeling in Transmission Electron MicroscopyI. Introduction; II. Sampling and Stereology; III. Quantities Displayed on Sections-Gold Labeling and Profile Data; IV. Quantities in Three Dimensions; V. Spatial Analysis; References; CHChapter 5: Combined Video Fluorescence and 3D Electron Microscopy; I. Introduction; II. Rationale; III. Method Steps; IV. Immunolabeling for EM with NANOGOLD; V. Immunolabeling for EM with HRP; References. CHChapter 6: From Live-Cell Imaging to Scanning Electron Microscopy (SEM): The Use of Green Fluorescent Protein (GFP) as a Common LabelI. Introduction; II. Rationale; III. Methods; IV. Summary; V. Concluding Remarks; References; CHChapter 7: Immunolabeling for Scanning Electron Microscopy (SEM) and Field Emission SEM; I. Introduction; II. Methods; III. Procedure; Conclusions; References; CHChapter 8: Immunogold Labeling of Thawed Cryosections; I. Introduction; II. Preparation of Carbon- and Formvar-Coated Copper Girds; III. Aldehyde Fixation of Cells for Cryoimmuno Labeling. IV. Embedding Samples for Cryoimmuno LabelingV. Cryosectioning for Cryoimmuno Labeling; VI. Immunogold Labeling; VII. Reagents and Solutions; VIII. Future Outlooks; References; CHChapter 9: Close-to-Native Ultrastructural Preservation by High Pressure Freezing; I. Introduction; II. Rational; III. Material; IV. Methods; V. High Pressure Frozen Samples; VI. Discussion; References; CHChapter 10: High-Pressure Freezing and Low-Temperature Fixation of Cell Monolayers Grown on Sapphire Coverslips; I. Introduction; II. Materials and Instrumentation; III. Procedures; IV. Comments and Pitfalls