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Correlative Light and Electron MIcroscopy
 
 

Correlative Light and Electron MIcroscopy, 1st Edition

 
Correlative Light and Electron MIcroscopy, 1st Edition,Thomas Mueller-Reichert,Paul Verkade,ISBN9780124160262
 
 
 

Methods in Cell Biology

Mueller-Reichert   &   Verkade   

Academic Press

9780124160262

9780123914385

460

235 X 191

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.

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Key Features

  • Covers many areas of CLEM by the best international scientists in the field
  • Includes a brief history and overview on CLEM methods
  • Description

    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.

    Readership

    Researchers and students in cell, molecular and developmental biology

    Thomas Mueller-Reichert

    Dr. Thomas Müller-Reichert is interested in how the microtubule cytoskeleton is modulated within cells to fulfill functions in meiosis, mitosis and abscission. The Müller-Reichert lab is mainly applying correlative light microscopy and electron tomography to study the 3D organization of microtubules in the early embryo of the nematode Caenorhabditis elegans and in tissue culture cells. He got his PhD degree from the Swiss Federal Institute of Technology (ETH) in Zurich and moved afterwards to the EMBL in Heidelberg (Germany) for a post-doc with Dr. Tony Hyman. He was a visiting scientist with Dr. Kent McDonald (UC Berkeley, USA) and set up the electron microscope facility at the newly founded Max Planck Institute of Molecular Cell Biology and Genetics (MPI-CBG). Since 2010 he is head of the Core Facility Cellular Imaging (CFCI) of the Medical Faculty Carl Gustav Carus of the TU Dresden (Germany). Together with Dr. Paul Verkade he has developed a rapid transfer system for high-pressure freezing used for Correlative Light and Electron Microcopy. He has organized a number microscopy conferences and taught in several (CL)EM courses. He edited an MCB volume on the Electron Microscopy of Model Systems.

    Affiliations and Expertise

    Core Facility Cellular Imaging, Faculty of Medicine Carl Gustav Carus, Dresden, Germany

    View additional works by Thomas Mueller-Reichert

    Paul Verkade

    Dr. Paul Verkade’s research focuses on the sorting mechanisms in intracellular transport pathways. His main tools are microscopy techniques, with an emphasis on electron microscopy (EM) in which field he has published over 50 papers. He has studied and got his PhD degree at the University of Utrecht, the Netherlands. After his post-doc time at the EMBL, Heidelberg, Germany in the group of Kai Simons and setting up a new EM lab at the Max Planck Institute for Molecular Cell Biology in Dresden, Germany he moved to the University of Bristol, UK in 2006. Here he set up a new EM unit as part of the Wolfson Bioimaging Facility, a fully integrated light and electron microscopy centre. To support his transport studies, part of his research is to develop techniques and tools for the use of Correlative Light Electron Microscopy (CLEM). Amongst other things he has developed the Rapid Transfer System for the EMPACT2 high-pressure freezer together with Leica Microsystems. This allows for the combination of time-resolved CLEM with optimal preservation of ultrastructure for EM. Dr. Verkade is chair of the Electron Microscopy section of the Royal Microscopical Society (RMS) and of the Cryo Microscopy Group, affiliated to the RMS. He has organised and taught on a large number of courses and workshops on subjects such as high-pressure freezing, Correlative Light Electron Microscopy, and immuno EM. He is also the principle organiser of the EMBO practical course on CLEM.

    Affiliations and Expertise

    Wolfson Bioimaging Facility, Schools of Biochemistry and Physiology & Pharmacology, University of Bristol, Bristol, UK

    Correlative Light and Electron MIcroscopy, 1st Edition

    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

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