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The combination of electron microscopy with transmitted light microscopy (termed correlative light and electron microscopy; CLEM) has been employed for decades to generate… Read more
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The combination of electron microscopy with transmitted light microscopy (termed correlative light and electron microscopy; CLEM) has been employed for decades to generate molecular identification that can be visualized by a dark, electron-dense precipitate. This new volume of Methods in Cell Biology covers many areas of CLEM, including a brief history and overview on CLEM methods, imaging of intermediate stages of meiotic spindle assembly in C. elegans embryos using CLEM, and capturing endocytic segregation events with HPF-CLEM.
Researchers and students in cell, molecular and developmental biology
Series Editors
Front Matter
Contributors
Preface
Introduction to Correlative Light and Electron Microscopy
Chapter 1 Imaging Fluorescently Labeled Complexes by Means of Multidimensional Correlative Light and Transmission Electron Microscopy
I Introduction
II Rationale
III Methods
IV Instrumentation and Materials
V Discussion
Chapter 2 Visualizing Live Dynamics and Ultrastructure of Intracellular Organelles with Preembedding Correlative Light-Electron Microscopy
I Introduction and Rationale
II Materials
III Methods
IV Discussion
V Summary
Chapter 3 Correlative Fluorescence and Transmission Electron Microscopy in Tissues
I Introduction
II Correlative Microscopy: Reporter Systems
III Correlative Microscopy of Tissues
IV Conclusions
Chapter 4 Correlative Light and Electron Microscopy in Parasite Research
I Introduction
II Rationale
III Methods
IV Experiments and Materials
V Results and Discussion
VI Summary
Chapter 5 Labeling of Ultrathin Resin Sections for Correlative Light and Electron Microscopy
I Introduction
II Rationale
III Methods
IV Materials
V Summary and Outlook
Chapter 6 3D HDO-CLEM
I Introduction
II Materials
III Methods
IV Notes
Chapter 7 Correlative Light and Electron Microscopy of GFP
I Introduction
II Rationale
III Methods
IV Materials
V Discussion
Chapter 8 Picking Faces out of a Crowd
I Introduction
II The Crowded Cell and Spatiotemporal Proteomics
III What EM has to Offer
IV Immunomarkers
V Genetically Appended or Inserted Protein Tags
VI Types of Genetic Tags Currently Available
VII Future Directions and Challenges
Chapter 9 Correlated Light Microscopy and Electron Microscopy
I Introduction
II Correlated Light and Electron Microscopy: Using the Best of Two Worlds
III ROIs: Search & Find Tools
IV Our Approach: Virtual Reality Overlay during Preparation
V Discussion and Conclusion
VI Future Perspective
Chapter 10 Capturing Endocytic Segregation Events with HPF-CLEM
I Introduction
II Methods
III Outlook
Chapter 11 Targeted Ultramicrotomy
I Introduction
II Rationale
III Methods
IV Instrumentation and Materials
V Discussion
Chapter 12 Correlative Light and Electron Microscopy of Intermediate Stages of Meiotic Spindle Assembly in the Early Caenorhabditis elegans Embryo
I Introduction
II Methods
III Instrumentation and Material
IV Discussion
Chapter 13 Precise, Correlated Fluorescence Microscopy and Electron Tomography of Lowicryl Sections Using Fluorescent Fiducial Markers
I Introduction
II Rationale
III Methods
IV Instrumentation and Materials
V Discussion
Chapter 14 Integrative Approaches for Cellular Cryo-electron Tomography
I Introduction
II Rationale
III Methods
IV Instrumentation and Materials
V Discussion and Outlook
Chapter 15 Visualizing Proteins in Electron Micrographs at Nanometer Resolution
I Introduction
II Rationale
III Methods
IV Instrumentations and Materials
V Discussion
VI Perspective
Chapter 16 Atmospheric Scanning Electron Microscope for Correlative Microscopy
IV Application Studies
V Discussion
Chapter 17 Bridging Microscopes
I Introduction
II Rationale
III Methods
IV Materials and Instrumentation
V Discussion
VI Summary
Chapter 18 Correlative Light and Volume Electron Microscopy
I Introduction
II Rationale
III Materials
IV Methods
V Discussion
VI Summary
Index
Volumes in Series
Colour Plate
TM
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