Correlative Light and Electron Microscopy

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Thomas Müller-Reichert, Paul Verkade
Academic Press, 2012 - Science - 404 pages
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The combination of electron microscopy with transmitted light microscopy (termed correlative light and electron microsİ 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.



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

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Contents

Chapter 2 Visualizing Live Dynamics and Ultrastructure of Intracellular Organelles with Preembedding Correlative LightElectron Microscopy
21
Chapter 3 Correlative Fluorescence and Transmission Electron Microscopy in Tissues
37
Chapter 4 Correlative Light and Electron Microscopy in Parasite Research
59
Chapter 5 Labeling of Ultrathin Resin Sections for Correlative Light and Electron Microscopy
75
Cellular Compartment Analysis by Correlative LightElectron Microscopy on Cryosection
95
Chapter 7 Correlative Light and Electron Microscopy of GFP
117
Genetic Labels for Identification of Proteins in Correlated Light and Electron Microscopy Imaging
139
Chapter 9 Correlated Light Microscopy and Electron Microscopy
157
Chapter 13 Precise Correlated Fluorescence Microscopy and Electron Tomography of Lowicryl Sections Using Fluorescent Fiducial Markers
235
Correlative Imaging and Focused Ion Beam Micromachining
259
Chapter 15 Visualizing Proteins in Electron Micrographs at Nanometer Resolution
283
Chapter 16 Atmospheric Scanning Electron Microscope for Correlative Microscopy
307
3D Correlative Light and Scanning Electron Microscopy of Complex Biological Structures
325
Using Focused Ion Beam Scanning Electron Microscopy to Image Transient Events in Model Organisms
357
Index
383
Volumes in Series
395

Chapter 10 Capturing Endocytic Segregation Events with HPFCLEM
175
A Valuable Tool for Correlated Light and Electron Microscopy of Small Model Organisms
203
Chapter 12 Correlative Light and Electron Microscopy of Intermediate Stages of Meiotic Spindle Assembly in the Early Caenorhabditis elegans Emb...
223
Colour Plates
405
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About the author (2012)

Dr. Thomas M ller-Reichert is interested in how the microtubule cytoskeleton is modulated within cells to fulfill functions in meiosis, mitosis and abscission. The M ller-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.

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.

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