Correlative Live Cell and Super Resolution Imaging of Autophagosome Formation

Methods Enzymol. 2017;587:1-20. doi: 10.1016/bs.mie.2016.09.050. Epub 2016 Nov 17.

Abstract

Autophagy is a highly dynamic intracellular process involving interactions between protein complexes and membranes. Direct observation of these components in living cells provides information on how they interact and when and where they are involved in the autophagy pathway. This chapter provides an overview of methods used to acquire images of fluorescently labeled components of the autophagy pathway in living cells using wide-field microscopy. Due to the diffraction-limited nature of this technique further details are provided on how to acquire postfixation correlative super resolution images from the same cells that have previously been imaged live. Combining these techniques offers an opportunity to follow the processes of autophagy in living cells with unprecedented detail.

Keywords: Autophagosome; Autophagy; Live imaging; Membrane dynamics; Super resolution.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Autophagosomes*
  • Autophagy
  • Cell Line
  • Humans
  • Image Processing, Computer-Assisted
  • Microscopy / methods*
  • Molecular Biology / instrumentation
  • Molecular Biology / methods*
  • Recombinant Proteins / analysis
  • Recombinant Proteins / genetics

Substances

  • Recombinant Proteins