Simultaneous assessment of autophagy and apoptosis using multispectral imaging cytometry

Autophagy. 2011 Sep;7(9):1045-51. doi: 10.4161/auto.7.9.16252. Epub 2011 Sep 1.

Abstract

Multiple stress pathways result in the induction of autophagy and apoptosis. Current methods (e.g., protein gel blot, microscopy) do not offer quantitative single-cell resolution, thus making it difficult to discern if these pathways are mutually exclusive or, in some situations, cooperative in executing cell death. We report a novel method that enables high-throughput, high-content assessment of LC3 puncta and caspase-3 cleavage at the single cell level.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Apoptosis* / drug effects
  • Apoptosis* / radiation effects
  • Autophagy* / drug effects
  • Autophagy* / radiation effects
  • Chloroquine / pharmacology
  • Embryo, Mammalian / cytology
  • Fibroblasts / cytology
  • Fibroblasts / drug effects
  • Fibroblasts / metabolism
  • Fibroblasts / radiation effects
  • Humans
  • Image Cytometry / methods*
  • Leukocytes, Mononuclear / cytology
  • Leukocytes, Mononuclear / drug effects
  • Leukocytes, Mononuclear / metabolism
  • Leukocytes, Mononuclear / radiation effects
  • Lysosomes / drug effects
  • Lysosomes / metabolism
  • Lysosomes / radiation effects
  • Mice
  • Microtubule-Associated Proteins / metabolism
  • Signal Transduction / drug effects
  • Signal Transduction / radiation effects
  • Time Factors
  • Ultraviolet Rays

Substances

  • Map1lc3b protein, mouse
  • Microtubule-Associated Proteins
  • Chloroquine