Tools to Measure Autophagy Using High Content Imaging and Analysis

Methods Mol Biol. 2018:1683:59-71. doi: 10.1007/978-1-4939-7357-6_5.

Abstract

Macroautophagy, hereafter referred to as autophagy, is a predominately pro-survival catabolic process responsible for the degradation of long-lived or aggregated proteins, invading microorganisms and damaged or redundant intracellular organelles. Removal of these entities is achieved through encompassment of the target by the autophagosome and subsequent delivery to the lysosome. The use of fluorescence microscopy is a common method to investigate autophagy through monitoring the spatial and temporal recruitment both of autophagosomal markers and cargo to the autophagosome. In this section, we will discuss the use of high content imaging (HCI) and analysis in the study of autophagy with reference to commonly used markers of autophagosomal formation.

Keywords: ATG5; Autophagosome; Autophagy; CRISPR; High content imaging; High content screening; LC3B.

MeSH terms

  • Animals
  • Autophagy*
  • Biomarkers
  • CRISPR-Cas Systems
  • Cells, Cultured
  • Fluorescent Antibody Technique
  • Gene Editing
  • Gene Knockout Techniques
  • Genes, Reporter
  • High-Throughput Screening Assays*
  • Humans
  • Image Processing, Computer-Assisted*
  • Mice
  • Microtubule-Associated Proteins / metabolism
  • Molecular Imaging* / methods
  • Phagosomes / metabolism
  • Rats

Substances

  • Biomarkers
  • Microtubule-Associated Proteins