Automated high-throughput high-content autophagy and mitophagy analysis platform

• Published in: Scientific reports Vol. 9; no. 1; pp. 9455 - 11
• Main Authors: Arias-Fuenzalida, Jonathan, Jarazo, Javier, Walter, Jonas, Gomez-Giro, Gemma, Forster, Julia I, Krueger, Rejko, Antony, Paul M A, Schwamborn, Jens C
• Format: Journal Article
• Language: English
• Published: England Nature Publishing Group 01.07.2019; Nature Publishing Group UK
• Subjects: Algorithms; Automation; Autophagosomes - metabolism; Autophagy; Autophagy - drug effects; Chloroquine - pharmacology; Homeostasis; Humans; Image processing; Image Processing, Computer-Assisted; Induced Pluripotent Stem Cells - cytology; Induced Pluripotent Stem Cells - metabolism; Intermediates; Lysosomes - metabolism; Microscopy, Fluorescence; Microtubule-Associated Proteins - genetics; Microtubule-Associated Proteins - metabolism; Mitophagy; Mitophagy - drug effects; Phagocytosis; Phenotyping
• ISSN: 2045-2322; 2045-2322
• DOI: 10.1038/s41598-019-45917-2

Autophagic processes play a central role in cellular homeostasis. In pathological conditions, the flow of autophagy can be affected at multiple and distinct steps of the pathway. Current analyses tools do not deliver the required detail for dissecting pathway intermediates. The development of new tools to analyze autophagic processes qualitatively and quantitatively in a more straightforward manner is required. Defining all autophagy pathway intermediates in a high-throughput manner is technologically challenging and has not been addressed yet. Here, we overcome those requirements and limitations by the developed of stable autophagy and mitophagy reporter-iPSC and the establishment of a novel high-throughput phenotyping platform utilizing automated high-content image analysis to assess autophagy and mitophagy pathway intermediates.