Correlative Light and Electron MIcroscopy

Correlative Light and Electron MIcroscopy

1st Edition - August 10, 2012
  • Editors: Thomas Mueller-Reichert, Paul Verkade
  • Hardcover ISBN: 9780124160262
  • eBook ISBN: 9780123914385

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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.

Key Features

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

Readership

Researchers and students in cell, molecular and developmental biology

Table of Contents

  • 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

Product details

  • No. of pages: 460
  • Language: English
  • Copyright: © Academic Press 2012
  • Published: August 10, 2012
  • Imprint: Academic Press
  • Hardcover ISBN: 9780124160262
  • eBook ISBN: 9780123914385

About the Serial Volume Editors

Thomas Mueller-Reichert

Thomas Mueller-Reichert
Thomas Müller-Reichert is a Professor of Structural Cell Biology at the Technische Universität Dresden (TU Dresden, Germany). He is interested in how the microtubule cytoskeleton is modulated within cells to fulfill functions in mitosis, meiosis and abscission. The Müller-Reichert lab is mainly applying correlative light microscopy and electron tomography to study the 3D organization of microtubules in early embryos and meiocytes of the nematode Caenorhabditis elegans, and also in mammalian cells in culture. He has published over 75 papers and edited several volumes of the Methods in Cell Biology series on electron microscopy and CLEM. TMR obtained his PhD at the Swiss Federal Institute of Technology (ETH) in Zurich and moved afterwards for a post-doc to the EMBL in Heidelberg (Germany). He was a visiting scientist with Dr. Kent McDonald (UC Berkeley, USA). Together with Paul Verkade, he 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 a scientific group leader and head of the Core Facility Cellular Imaging (CFCI) of the Faculty of Medicine Carl Gustav Carus of the TU Dresden. He acted as president of the German Society for Electron Microscopy (Deutsche Gesellschaft für Elektronenmikroskopie, DGE) from 2018 to 2019. He taught numerous courses and workshops on high-pressure freezing and Correlative Light and Electron Microscopy.

Affiliations and Expertise

Professor of Structural Cell Biology, Technische Universitat Dresden, TU Dresden, Germany

Paul Verkade

Paul Verkade
Paul Verkade is a Professor of Bioimaging at the University of Bristol, UK where his research group works on the development and application of microscopy techniques to Biomedical questions. His focus is on the study of sorting mechanisms in intracellular transport pathways and in the area of Synthetic Biology. The main tools in the lab are Electron microscopy (EM) and Correlative Light Electron Microscopy (CLEM) in which fields he has published over 85 papers and edited 4 books on CLEM (including 3 Volumes of the Methods in Cell Biology series). PV obtained his PhD on electron microscopic studies of the peripheral nervous system at the University of Utrecht, The Netherlands in 1996. Subsequently he did a post-doc at the EMBL, Heidelberg, Germany, after which he set up the electron microscopy unit at the newly formed Max Planck Institute for Molecular Cell Biology in Dresden, Germany from 2001. He moved to the UK in 2006 to set up another EM unit as part of an integrated LM and EM bioimaging facility, which facilitates CLEM workflows. He has acted as chair and co-chair of the Electron Microscopy section of the Royal Microscopical Society and is closely involved with BioimagingUK shaping the UK imaging infrastructure landscape. He has organised and taught on several courses and workshops on subjects such as high-pressure freezing, Correlative Light Electron Microscopy, and immuno EM. His lab is the home of the EMBO practical course on CLEM and he is Work Group leader for CLEM within the EU COST project COMULIS (COrrelated MUltimodal imaging in LIfe Sciences).

Affiliations and Expertise

Professor of Bioimaging, University of Bristol, UK