Correlative light and electron microscopy III
Front Cover -- Methods in Cell Biology -- Series Editors -- Methods in CellBiology: Correlative Light and Electron Microscopy III -- Copyright -- Contents -- Contributors -- Preface: The Key Lies in the Right Combination -- 1 - Millisecond time resolution correlative light and electron microscopy fo...
Gespeichert in:
| Weitere Verfasser: | , |
|---|---|
| Dokumenttyp: | Book/Monograph |
| Sprache: | Englisch |
| Veröffentlicht: |
Amsterdam Boston London
Elsevier Science
[2017]
|
| Ausgabe: | First edition |
| Schriftenreihe: | Methods in cell biology
volume 140 |
| In: |
Methods in cell biology (volume 140)
|
| Volumes / Articles: | Show Volumes / Articles. |
| Online-Zugang: | Verlag, Volltext: http://www.sciencedirect.com/science/bookseries/0091679X/140
|
| Verfasserangaben: | edited by Thomas Müller-Reichert and Paul Verkade |
| Zusammenfassung: | Front Cover -- Methods in Cell Biology -- Series Editors -- Methods in CellBiology: Correlative Light and Electron Microscopy III -- Copyright -- Contents -- Contributors -- Preface: The Key Lies in the Right Combination -- 1 - Millisecond time resolution correlative light and electron microscopy for dynamic cellular processes -- INTRODUCTION -- 1. METHODS -- 1.1 Chlamydomonas reinhardtii Culture -- 1.2 Imaging and Embedding Chamber Preparation -- 1.3 Light Microscopy -- 1.4 Sample Preparation for Electron Microscopy -- 1.4.1 Chemical postfixation and embedding -- 1.4.2 Thin sectioning -- 1.4.3 Staining of the sections and application of fiducial markers -- 1.5 Electron Tomography -- 1.6 Registration of Light and Electron Microscopy Images -- 1.7 Data Analysis -- 1.8 Preparation of Flagellar Cross Sections for Correlative Light and Electron Microscopy Analysis -- 2. INSTRUMENTATION AND MATERIALS -- 2.1 Chlamydomonas reinhardtii culture -- 2.2 Imaging and Embedding Chamber Preparation -- 2.3 Light Microscopy -- 2.4 Sample Preparation for Electron Microscopy -- 2.4.1 Chemical postfixation and embedding -- 2.4.2 Thin sectioning -- 2.4.3 Staining of the sections and application of fiducial markers -- 2.5 Electron Tomography -- 2.6 Registration of Light and Electron Microscopy Images -- 2.7 Data Analysis -- 2.8 Preparation of Flagellar Cross Sections for Correlative Light and Electron Microscopy Analysis -- 3. DISCUSSION AND OUTLOOK -- Acknowledgments -- REFERENCES -- 2 - 3D subcellular localization with superresolution array tomography on ultrathin sections of various species -- INTRODUCTION AND RATIONALE -- 1. METHODS-CORE PROTOCOL -- 1.1 High-Pressure Freezing -- 1.2 Freeze Substitution -- 1.3 Embedding -- 1.4 Ultramicrotomy -- 1.5 Light Microscopy -- 1.6 Contrasting and Carbon Coating -- 1.7 Scanning Electron Microscopy 1.8 Image Processing and Correlation of Structured Illumination Microscopy and Scanning Electron Microscopy Images -- 1.9 Alignment and 3D Reconstruction -- 1.9.1 IMOD -- 1.9.2 Fiji and AMIRA -- 2. METHODS-MODEL-SPECIFIC ADAPTATIONS AND CONSIDERATIONS -- 2.1 Caenorhabditis elegans -- 2.2 Trypanosoma brucei -- 2.3 Social Insects (Apis mellifera and Cataglyphis fortis) -- 3. INSTRUMENTATION AND MATERIALS -- 3.1 High-Pressure Freezing -- 3.1.1 General -- 3.1.2 Caenorhabditis elegans -- 3.1.3 Trypanosoma brucei -- 3.1.4 Cataglyphis fortis and Apis mellifera -- 3.2 Freeze Substitution -- 3.3 Embedding -- 3.4 Ultramicrotomy -- 3.5 Light Microscopy -- 3.5.1 Primary Antibodies -- 3.6 Contrasting and Carbon Coating -- 3.7 Scanning Electron Microscopy -- 3.8 Image Processing and Correlation of Structured Illumination Microscopy and Scanning Electron Microscopy Images -- 3.9 Alignment and 3D Reconstruction -- CONCLUSIONS -- Acknowledgments -- REFERENCES -- 3 - Preserving the photoswitching ability of standard fluorescent proteins for correlative in-resin super-resolutio ... -- INTRODUCTION -- 1. RATIONALE -- 2. MATERIALS -- 2.1 Instrumentation -- 2.2 Materials -- 2.3 Chemicals -- 3. METHODS -- 3.1 Reagent Preparation -- 3.2 High Pressure Freezing and Freeze Substitution -- 3.3 Resin Infiltration and Polymerization -- 3.4 Ultramicrotomy -- 3.5 Single Molecule Localization Microscopy -- 3.6 Transmission Electron Microscopy -- 3.7 Correlation of Single Molecule Localization Microscopy and Transmission Electron Microscopy Images -- 4. DISCUSSION -- 4.1 Preservation of Fluorophore Photoswitching -- 4.2 Applications of the Technique -- 4.3 Limitations -- 4.4 Future Prospects -- Acknowledgments -- REFERENCES -- 4 - Minimal resin embedding of multicellular specimens for targeted FIB-SEM imaging -- INTRODUCTION -- 1. RATIONALE -- 2. METHODS 2.1 From Immobilization to Infiltration -- 2.1.1 Chemical fixation -- 2.1.1.1 Zebrafish embryos (Danio rerio) -- 2.1.1.2 Platynereis (Platynereis dumerilii) -- 2.1.2 High-pressure freezing -- 2.1.2.1 Caenorhabditis elegans dauer larvae -- 2.2 Minimal Resin -- 3. RESULTS: FIB-SEM TARGETING AND IMAGING -- 4. INSTRUMENTATION AND MATERIALS -- 4.1 Chemical Fixation -- 4.2 High-Pressure Freezing -- 5. DISCUSSION -- Acknowledgments -- REFERENCES -- 5 - A new method for cryo-sectioning cell monolayers using a correlative workflow -- INTRODUCTION -- 1. RATIONALE -- 2. METHODS -- 2.1 Cell Culture on Finder Grids -- 2.1.1 Preparation of the finder grids -- 2.1.2 Cell culture for whole-mount correlation -- 2.2 Correlative Light and Electron Microscopy Option A: Live-Cell Fluorescence Microscopy Before High-Pressure Freezing -- 2.3 High-Pressure Freezing -- 2.4 Correlative Light and Electron Microscopy Option B: Cryo-Fluorescence Microscopy After High-Pressure Freezing -- 2.5 Mounting of the Carrier Into the Cryo-Ultramicrotome -- 2.6 Correlative Light and Electron Microscopy Option C: Fluorescence Targeting at the Microtome -- 2.7 Trimming -- 2.8 Sectioning -- 2.9 Correlative Light and Electron Microscopy Option D: Cryo-Correlative Light and Electron Microscopy -- 3. INSTRUMENTATION, MATERIALS, AND REAGENTS -- 3.1 Instruments -- 3.2 Materials and Reagents -- 4. RESULTS AND DISCUSSION -- Acknowledgments -- REFERENCES -- 6 - Correlative light and electron microscopic detection of GFP-labeled proteins using modular APEX -- 1. VECTORS FOR CORRELATIVE LIGHT AND ELECTRON MICROSCOPY IN MAMMALIAN CELL CULTURE AND WHOLE ZEBRAFISH -- 2. CORRELATIVE LIGHT AND ELECTRON MICROSCOPY IN CELL CULTURE -- 2.1 Method -- 2.1.1 Cell culture and transient transfections -- 2.1.2 Transfections -- 2.1.3 Light and fluorescence microscopy 2.1.4 Sample processing for transmission electron microscopy -- 2.1.5 Transmission electron microscopy -- 2.1.6 Postimage processing -- 2.2 Materials and Instrumentation -- 2.2.1 Cell culture -- 2.2.2 Light microscopy -- 2.2.3 Electron microscopy -- 2.2.4 Image processing -- 3. SUBCELLULAR PROTEIN DISTRIBUTION ANALYSIS OF TRANSGENIC ZEBRAFISH -- 3.1 Methods -- 3.1.1 Zebrafish crossing -- 3.1.2 Mounting fish samples for confocal microscopy -- 3.1.3 Sample processing for transmission electron microscopy -- 3.1.4 Transmission electron microscopy -- 3.1.5 Postimage processing -- 3.2 Materials and Instrumentation -- 3.2.1 Subcellular protein distribution analysis of transgenic zebrafish -- 3.2.1.1 Zebrafish -- 3.2.1.2 Confocal microscopy -- 3.2.1.3 Electron microscopy -- 3.2.1.4 Image processing -- 3.3 Discussion -- REFERENCES -- 7 - Correlation of live-cell imaging with volume scanning electron microscopy -- INTRODUCTION -- 1. RATIONALE -- 2. METHODS -- 2.1 Choice of Cell Culture Substrates -- 2.1.1 Flow-chamber setup -- 2.1.2 Cell culture dishes with gridded glass substrate -- 2.2 Cell Culture -- 2.3 Live-Cell Imaging, Fixation, and 3D Imaging -- 2.4 Sample Preparation for Electron Microscopy -- 2.4.1 Staining and dehydration -- 2.4.2 En-bloc embedding -- 2.4.3 Thin-layer plastification -- 2.4.4 Specimen mounting for volume SEM -- 2.5 Relocation of Cells of Interest and Volume SEM Imaging -- 2.5.1 Focused ion beam-scanning electron microscopy -- 2.5.2 Serial blockface-scanning electron microscopy -- 2.6 Visualizing the Correlation -- 3. MATERIALS -- 3.1 Equipment -- 3.2 Consumables and Chemicals -- 4. DISCUSSION -- 4.1 Live-Cell Imaging Setup -- 4.2 The Influence of Sample Preparation -- 4.3 Relocation of the Structure of Interest and Correlation of LM and EM Data -- CONCLUSIONS -- Acknowledgments -- REFERENCES 8 - A fully integrated, three-dimensional fluorescence to electron microscopy correlative workflow -- INTRODUCTION -- 1. MATERIALS AND METHODS -- Molecular Biology -- Day 1: Cell culture and transfection -- Day 2: µ-Slide cell seeding -- Day 3: FEI CorrSight Live cell imaging -- Day 4: Electron microscopy sample preparation -- Day 5: Mounting and coating blocks for electron microscopy -- Relocalization of cell of interest in the DualBeam -- Preparation of the cell of interest for ASV acquisition -- Day 9: Image alignment and segmentation using Amira -- 2. DISCUSSION -- 3. SUMMARY -- Acknowledgments -- SUPPLEMENTARY DATA -- REFERENCES -- 9 - CLAFEM: correlative light atomic force electron microscopy -- INTRODUCTION -- 1. MATERIALS AND METHODS -- 1.1 Cell Culture -- 1.2 Sample Preparation-Live-Imaging of Actin Cytoskeleton -- 1.3 Sample Preparation-LC-3 Positive Yersinia Vacuoles and Microtubules -- 1.4 Correlation Technique -- 1.5 Atomic Force Microscopy -- 1.6 Stimulated Emission Depletion Microscopy -- 1.7 Transmission Electron Microscopy -- 1.8 Scanning Electron Microscopy -- 2. RESULTS AND DISCUSSION -- Acknowledgments -- REFERENCES -- 10 - Correlative light-electron microscopy in liquid using an inverted SEM (ASEM) -- INTRODUCTION -- 1. INSTRUMENT DESIGN AND SAMPLE GEOMETRY OF THE ATMOSPHERIC SCANNING ELECTRON MICROSCOPY -- 1.1 Configuration of the Atmospheric Scanning Electron Microscopy -- 1.2 The Atmospheric Scanning Electron Microscopy Dish -- 1.3 Column Protection System in Case of Accidental SiN Film Breakage -- 1.4 Labeling and Staining for Correlative Light-Electron Microscopy -- 1.5 Immunolabeling Method for Correlative Light-Electron Microscopy -- 2. THE APPLICATION OF CORRELATIVE LIGHT-ELECTRON MICROSCOPY USING ATMOSPHERIC SCANNING ELECTRON MICROSCOPY -- 2.1 Endoplasmic Reticulum 2.2 Super Molecular Complex Formation of STIM1 by Sensing Ca2+ |
|---|---|
| Beschreibung: | Online Resource |
| ISBN: | 9780128099766 |