Quantifying Autophagic Structures in Mammalian Cells Using Confocal Microscopy

doi:10.1016/bs.mie.2016.09.051

. 2017:587:21-42.

doi: 10.1016/bs.mie.2016.09.051. Epub 2016 Nov 12.

Quantifying Autophagic Structures in Mammalian Cells Using Confocal Microscopy

C A Lamb¹, J Joachim², S A Tooze³

Affiliations

¹ Molecular Cell Biology of Autophagy Group, The Francis Crick Institute, London, United Kingdom. Electronic address: christopheralamb101@gmail.com.
² Molecular Cell Biology of Autophagy Group, The Francis Crick Institute, London, United Kingdom.
³ Molecular Cell Biology of Autophagy Group, The Francis Crick Institute, London, United Kingdom. Electronic address: sharon.tooze@crick.ac.uk.

PMID: 28253957
DOI: 10.1016/bs.mie.2016.09.051

Quantifying Autophagic Structures in Mammalian Cells Using Confocal Microscopy

C A Lamb et al. Methods Enzymol. 2017.

. 2017:587:21-42.

doi: 10.1016/bs.mie.2016.09.051. Epub 2016 Nov 12.

Authors

C A Lamb¹, J Joachim², S A Tooze³

Affiliations

¹ Molecular Cell Biology of Autophagy Group, The Francis Crick Institute, London, United Kingdom. Electronic address: christopheralamb101@gmail.com.
² Molecular Cell Biology of Autophagy Group, The Francis Crick Institute, London, United Kingdom.
³ Molecular Cell Biology of Autophagy Group, The Francis Crick Institute, London, United Kingdom. Electronic address: sharon.tooze@crick.ac.uk.

PMID: 28253957
DOI: 10.1016/bs.mie.2016.09.051

Abstract

Autophagy relies on the sequential, hierarchical association of proteins with phagophores, and forming autophagosomes to allow completion of the process. Additionally, the trafficking of the unique transmembrane autophagy-related protein ATG9 is vital for autophagy progression. In this chapter, we discuss methods to monitor autophagosome number using confocal microscopy, by following the association of different autophagosomal markers with the phagophore and completed autophagosome. We also discuss methods to monitor the trafficking of ATG9 in mammalian cells under starvation conditions.

Keywords: ATG9; Autophagosome; Autophagy; Imaris; LC3; Live cell imaging.

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Quantifying Autophagic Structures in Mammalian Cells Using Confocal Microscopy

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Quantifying Autophagic Structures in Mammalian Cells Using Confocal Microscopy

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