Super-resolution correlative light-electron microscopy using a click-chemistry approach for studying intracellular trafficking
| Autor: | Andrian, T., Bakkum, T., Elsland, D.M. van, Bos, E., Koster, A.J., Albertazzi, L., Kasteren, S.I. van, Pujals, S., MullerReichert, T., Verkade, P. |
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| Přispěvatelé: | Müller-Reichert, T., Verkade, P., Molecular Biosensing for Med. Diagnostics, ICMS Core |
| Jazyk: | angličtina |
| Rok vydání: | 2020 |
| Předmět: |
0303 health sciences
Super-resolution microscopy Correlative light and electron microscopy Resolution (electron density) Context (language use) Nanotechnology Biology Tokuyasu cryo-sectioning law.invention 03 medical and health sciences law Transmission electron microscopy Click-chemistry Microscopy Ultrastructure Electron microscope Single molecule localization microscopy Nanoscopic scale 030304 developmental biology STochastic Optical Reconstruction Microscopy (STORM) |
| Zdroj: | Methods in Cell Biology, 162, 303-331. Elsevier Methods in Cell Biology Methods in Cell Biology-Correlative Light and Electron Microscopy IV Correlative Light and Electron Microscopy IV, 303-331 STARTPAGE=303;ENDPAGE=331;TITLE=Correlative Light and Electron Microscopy IV Methods in Cell Biology, 162, 303-331 Correlative Light and Electron Microscopy IV Methods in Cell Biology ISBN: 9780128220580 |
| ISSN: | 0091-679X |
| Popis: | Correlative light and electron microscopy (CLEM) entails a group of multimodal imaging techniques that are combined to pinpoint to the location of fluorescently labeled molecules in the context of their ultrastructural cellular environment. Here we describe a detailed workflow for STORM-CLEM, in which STochastic Optical Reconstruction Microscopy (STORM), an optical super-resolution technique, is correlated with transmission electron microscopy (TEM). This protocol has the advantage that both imaging modalities have resolution at the nanoscale, bringing higher synergies on the information obtained. The sample is prepared according to the Tokuyasu method followed by click-chemistry labeling and STORM imaging. Then, after heavy metal staining, electron microscopy imaging is performed followed by correlation of the two images. The case study presented here is on intracellular pathogens, but the protocol is versatile and could potentially be applied to many types of samples. |
| Databáze: | OpenAIRE |
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