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

Details

No. of pages:
460
Language:
English
Copyright:
© 2012
Published:
Imprint:
Academic Press
Electronic ISBN:
9780123914385
Print ISBN:
9780124160262

About the serial-volume-editors

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.

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.