Correlative Light and Electron Microscopy of Autophagosomes
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Live-cell imaging has been widely used to study autophagosome biogenesis and maturation. When combined with correlative electron microscopy, this approach can be extended to reveal ultrastructural details in three dimensions. The resolution of electron microscopy is needed when membrane contact sites and tubular connections between organelles are studied.
Key wordsLive-cell imaging Phagophore Autophagosome Serial sectioning Electron tomography
Authors’ laboratory is supported by the Academy of Finland and Magnus Ehrnrooth Foundation. Live-cell imaging and confocal microscopy were carried out in the Light Microscopy Unit at the Institute of Biotechnology, University of Helsinki. We thank the Electron Microscopy Unit at the Institute of Biotechnology, University of Helsinki, for technical help in thin sectioning and for the possibility to use an electron microscope. HeLa cells stably expressing mRFP-GFP-LC3 were a kind gift from Tamotsu Yoshimori, University of Osaka, Japan, and HEK293 cells stably expressing ATG13 were a kind gift from Nicholas Ktistakis, Babraham Institute, Cambridge, UK.
- 5.Killingsworth MC, Bobryshev YV (2016) Correlative light- and electron microscopy using quantum dot nanoparticles. J Vis Exp. https://doi.org/10.3791/54307