High throughput screening for autophagy

Methods Cell Biol. 2021:165:89-101. doi: 10.1016/bs.mcb.2020.12.011. Epub 2021 Jan 23.

Abstract

Robotized high throughput screening allows for the assessment of autophagy in a large number of samples. Here, we describe a drug discovery platform for the phenotypic identification of novel autophagy inducers by means of automated cell biology workflows employing robotized cell culture, sample preparation and data acquisition. In this setting, fluorescent biosensor cells that express microtubule-associated proteins 1A/1B light chain 3B (best known as LC3) conjugated to green fluorescent protein (GFP), are utilized together with automated high content microscopy for the image-based assessment of autophagy. In sum, we detail a drug discovery screening workflow from high throughput sample preparation and processing to data acquisition and analysis.

Keywords: Automation; Cancer; Drug discovery; Imaging.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Autophagy*
  • Drug Discovery
  • Drug Evaluation, Preclinical
  • Green Fluorescent Proteins / genetics
  • High-Throughput Screening Assays*
  • Microtubule-Associated Proteins

Substances

  • Microtubule-Associated Proteins
  • Green Fluorescent Proteins