Summary
This chapter describes the electron microscopic fine structure of early and late autophagic vacuoles in mammalian cells. Detailed instructions are given for the preparation of cells for conventional electron microscopy and for the identification of autophagic vacuoles by morphology. Electron microscopy remains one of the most accurate methods for quantitation of autophagic vacuole accumulation. Therefore, quantitation of autophagic vacuoles by electron microscopy and point counting is also described. Finally, a short description is given for preparation of ultra thin cryosections for immunogold labeling of autophagic vacuoles.
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Acknowledgments
The author would like to thank Pirkko Hirsimäki (Turku, Finland) and Hilkka Reunanen (Jyväskylä, Finland) for the initial introduction to the secrets of electron microscopy and autophagy. John Lucocq (Dundee, UK) is thanked for sharing his knowledge on the quantitative aspects of microscopy, and Paul Saftig (Kiel, Germany) is acknowledged for his generous support during the years we have used to explore the functions of lysosomal membrane proteins. Work described in this chapter was supported by The Royal Society (Dundee), Helsel Stiftung (Kiel), University of Helsinki Research Funds, and Biocentrum Helsinki
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Eskelinen, EL. (2008). Fine Structure of the Autophagosome. In: Deretic, V. (eds) Autophagosome and Phagosome. Methods in Molecular Biology™, vol 445. Humana Press. https://doi.org/10.1007/978-1-59745-157-4_2
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DOI: https://doi.org/10.1007/978-1-59745-157-4_2
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