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Ultrastructural Localization of Endogenous LC3 by On-Section Correlative Light-Electron Microscopy
JoVE Journal
Biologie
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JoVE Journal Biologie
Ultrastructural Localization of Endogenous LC3 by On-Section Correlative Light-Electron Microscopy

Ultrastructural Localization of Endogenous LC3 by On-Section Correlative Light-Electron Microscopy

DOI:
10.3791/65067-v

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11:53 min

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March 31, 2023

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, , , , , ,
1Center for Molecular Medicine-Cell Biology, University Medical Center Utrecht,Utrecht University

Kapitel

  • 00:05Introduction
  • 00:50Sample Embedding
  • 03:13Sectioning
  • 04:41Labeling and Light Microscopy
  • 09:18Results: Correlative Light-EM of LC3 and LAMP1 in Starved HEPG2 Cells
  • 11:16Conclusion

Summary

Automatische Übersetzung

Automatische Übersetzung

Here, we present a protocol for optimized on-section correlative light-electron microscopy based on endogenous, fluorescent labeling as a tool to investigate the localization of rare proteins in relation to cellular ultrastructure. The power of this approach is demonstrated by ultrastructural localization of endogenous LC3 in starved cells without Bafilomycin treatment.

Tags

Ultrastructural LocalizationCorrelative Light-electron MicroscopyAutophagy ProteinsFluorescence MicroscopyElectron MicroscopyOn-section CLEMCell UltrastructureGelatin EmbeddingSucrose CryoprotectionCryosectioning

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