Correlative Light and Electron Microscopy IV

1st Edition, Volume 162 - March 8, 2021
Imprint: Academic Press
Editors: Thomas Muller-Reichert, Paul Verkade
Language: English
Hardback ISBN:
9 7 8 - 0 - 1 2 - 8 2 2 0 5 8 - 0
eBook ISBN:
9 7 8 - 0 - 1 2 - 8 2 2 0 5 9 - 7

Correlative Light and Electron Microscopy IV, Volume 162, a new volume in the Methods in Cell Biology series, continues the legacy of this premier serial with quality chapters auth… Read more

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Correlative Light and Electron Microscopy IV, Volume 162, a new volume in the Methods in Cell Biology series, continues the legacy of this premier serial with quality chapters authored by leaders in the field. Besides the detailed description of protocols for CLEM technologies including time-resolution, Super resolution LM and Volume EM, new chapters cover Workflow (dis)-advantages/spiderweb, Serial section LM + EM, Platinum clusters as CLEM probes, Correlative Light Electron Microscopy with a transition metal complex as a single probe, SEM-TEM-SIMS, HPF-CLEM, A new workflow for high-throughput screening of mitotic mammalian cells for electron microscopy using classic histological dyes, and more.

Cover image
Title page
Table of Contents
Copyright
Contributors
Preface to CLEM IV: Broaden the horizon
What does it hold for you?
Chapter 1: 50 Shades of CLEM: How to choose the right approach for you
Abstract
1: What is CLEM?
2: The microscopist's dilemma
3: Sample preservation
4: Case studies
5: How to choose your shade of CLEM: Closing remarks
Acknowledgments
Chapter 2: The Histo-CLEM Workflow for tissues of model organisms
Abstract
1: Introduction
2: General design of the Histo-CLEM Workflow
3: Application example
4: Concluding remarks
Acknowledgments
Chapter 3: Fluorescent platinum nanoclusters as correlative light electron microscopy probes
Abstract
1: Introduction
2: CLEM probes
3: Instrumentation, materials, and reagents
4: Methods
5: Generation and validation of the probe
6: Use of PtNCs
7: Discussion and outlook
Acknowledgments
Chapter 4: Refining a correlative light electron microscopy workflow using luminescent metal complexes
Abstract
1: Introduction
2: Transition metal Ir complex 1
3: Instrumentation and materials
4: Methods
5: Results
6: Discussion
Acknowledgments
Chapter 5: Sample preparation for energy dispersive X-ray imaging of biological tissues
Abstract
1: Introduction
2: Routine chemical fixation of tissues
3: Resin
4: Contrasting
5: Grids and sample holder
6: Other considerations
7: Conclusion
8: Materials
Acknowledgments
Author contributions
Chapter 6: HPM live μ for a full CLEM workflow
Abstract
1: Introduction
2: High pressure freezing principles
3: Designing the HPM live μ
4: Integration of the HPM live μ in a CLEM workflow
5: A live imaging, cryo-fluorescence, in-resin fluorescence and EM workflow: Validation at each step
6: Perspectives
Acknowledgments
Author contributions
Chapter 7: High-throughput screening of mitotic mammalian cells for electron microscopy using classic histological dyes
Abstract
1: Introduction
2: Methods
3: Instrumentation and materials
4: Discussion
Acknowledgments
Chapter 8: On-section correlative light and electron microscopy of large cellular volumes using STEM tomography
Abstract
1: Introduction
2: Methods
3: Instrumentation and materials
4: Results
5: Discussion
6: Conclusion
Acknowledgments
Chapter 9: An accelerated procedure for approaching and imaging of optically branded region of interest in tissue
Abstract
1: Introduction
2: Rationale
3: Methods
4: Instrumentation and materials
5: Discussion and conclusion
Acknowledgments
Chapter 10: Cryo-fluorescence microscopy of high-pressure frozen C. elegans enables correlative FIB-SEM imaging of targeted embryonic stages in the intact worm
Abstract
1: Introduction
2: A note on selecting the right cryoprotectant
3: Rationale
4: Methods
5: Instrumentation and materials
6: Discussion
Acknowledgments
Chapter 11: Correlative super-resolution fluorescence and electron cryo-microscopy based on cryo-SOFI
Abstract
1: Introduction
2: Rationale
3: Materials
4: Methods
5: Discussion
Acknowledgments
Chapter 12: Cryo-correlative light and electron microscopy workflow for cryo-focused ion beam milled adherent cells
Abstract
1: Introduction
2: Rationale
3: Materials
4: Methods
5: Results and discussion
Acknowledgments
Chapter 13: Super-resolution correlative light-electron microscopy using a click-chemistry approach for studying intracellular trafficking
Abstract
1: Introduction
2: Methods
3: Instrumentation and materials
4: Discussion
Acknowledgments
Chapter 14: Step-by-step guide to post-acquisition correlation of confocal and FIB/SEM volumes using Amira software
Abstract
1: Introduction
2: Precursor techniques
3: Software and hardware
4: Step-by-step guide
5: Summary and discussion
Acknowledgments
Chapter 15: Visualization and co-registration of correlative microscopy data with the ImageJ plug-in Correlia
Abstract
1: Introduction
2: Models for image co-registration
3: Correlia
4: Two real-life examples
5: Summary and conclusions
Acknowledgments
Chapter 16: Multimodality imaging beyond CLEM: Showcases of combined in-vivo preclinical imaging and ex-vivo microscopy to detect murine mural vascular lesions
Abstract
1: Introduction
2: Rationale
3: Methods and results
4: Discussion
5: Conclusion and outlook
Acknowledgment
Chapter 17: Correlative multimodal imaging: Building a community
Abstract
1: Setting the scene
2: A brief history
3: Building the community
4: Conclusion
Acknowledgments

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, Germany

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

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