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Correlative Light and Electron MIcroscopy
1st Edition - August 10, 2012
Editors: Thomas Muller-Reichert, Paul Verkade
Hardback ISBN:
9 7 8 - 0 - 1 2 - 4 1 6 0 2 6 - 2
eBook ISBN:
9 7 8 - 0 - 1 2 - 3 9 1 4 3 8 - 5
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.
- Covers many areas of CLEM by the best international scientists in the field
- Includes a brief history and overview on CLEM methods
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
- No. of pages: 460
- Language: English
- Published: August 10, 2012
- Imprint: Academic Press
- Hardback ISBN: 9780124160262
- eBook ISBN: 9780123914385
TM
Thomas Muller-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, GermanyPV
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. The main tools in the lab are Electron microscopy (EM) and Correlative Light Electron Microscopy (CLEM) in which fields he has published over 100 papers and edited 5 books on CLEM (including 4 Volumes of the Methods in Cell Biology series). PV obtained his PhD 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 is actively involved in shaping the future microscopy landscape with roles in the Royal Microscopical Society and BioimagingUK and a current focus on putting volumeEM on the imaging map through community building and the organisation and co-chairing of the 1st Gordon Research Conference on vEM.
Affiliations and expertise
The University of Bristol, Bristol, UK