High-Content Autophagy Analysis in iPSC-Derived Neurons Using Immunofluorescence

Methods Mol Biol. 2019:1994:165-174. doi: 10.1007/978-1-4939-9477-9_15.

Abstract

Autophagy is the process by which cellular proteins and organelles are degraded and recycled and is essential to the survival of cells. Defective autophagic degradation has been linked to many neurodegenerative diseases and in particular lysosomal storage diseases. Here we describe a high-content assay to detect defects in the autophagy pathway in induced pluripotent stem cell-derived neurons. This assay utilizes immunofluorescence to stain autophagosomes and uses automated image analysis to measure changes in autophagosome levels in response to modulators of autophagy.

Keywords: Autophagy; High-content screening; Image analysis; Induced pluripotent stem cells; Neurons.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Autophagosomes / metabolism*
  • Autophagy / physiology*
  • Fluorescent Antibody Technique / methods*
  • Humans
  • Image Processing, Computer-Assisted
  • Induced Pluripotent Stem Cells / cytology*
  • Microscopy, Fluorescence
  • Neurons / cytology*
  • Neurons / metabolism