Automated Detection of Autophagy Response Using Single Cell-Based Microscopy Assays

Methods Mol Biol. 2019;1880:429-445. doi: 10.1007/978-1-4939-8873-0_27.

Abstract

The fluorescence microscopy-based detection of intracellular LC3, p62, and/or WIPI punctate structures is a robust tool to monitor and assess macroautophagy/autophagy in single cells. This method was established for automated high-throughput/content analysis to reliably detect narrow differences in autophagy activity/capacity and to provide screening opportunities for biological and chemical libraries.

Keywords: Automated high-throughput analysis; Autophagy; LC3; WIPI; p62.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Autophagy / physiology*
  • Autophagy-Related Proteins / metabolism
  • Cell Culture Techniques / instrumentation
  • Cell Culture Techniques / methods
  • Cell Line
  • High-Throughput Screening Assays / instrumentation
  • High-Throughput Screening Assays / methods*
  • Humans
  • Image Processing, Computer-Assisted / instrumentation
  • Image Processing, Computer-Assisted / methods*
  • Microscopy, Confocal / instrumentation
  • Microscopy, Confocal / methods
  • Microscopy, Fluorescence / instrumentation
  • Microscopy, Fluorescence / methods
  • Microtubule-Associated Proteins / metabolism
  • Single-Cell Analysis / instrumentation
  • Single-Cell Analysis / methods*

Substances

  • Autophagy-Related Proteins
  • Microtubule-Associated Proteins