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Tools to Measure Autophagy Using High Content Imaging and Analysis.

Nick J Dolman1, Brent A Samson1, Kevin M Chambers2

  • 1Thermo Fisher Scientific, 100 Technology Drive, Pittsburgh, PA, 15219, USA.

Methods in Molecular Biology (Clifton, N.J.)
|October 31, 2017
PubMed
Summary
This summary is machine-generated.

Autophagy, a cellular degradation process, removes damaged components and pathogens. High content imaging (HCI) aids in studying autophagosome formation and markers, offering insights into this vital survival mechanism.

Keywords:
ATG5AutophagosomeAutophagyCRISPRHigh content imagingHigh content screeningLC3B

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Area of Science:

  • Cell Biology
  • Molecular Biology
  • Biochemistry

Background:

  • Macroautophagy (autophagy) is a fundamental catabolic process crucial for cellular homeostasis.
  • It involves the degradation of long-lived proteins, aggregated proteins, invading microorganisms, and damaged organelles.
  • Autophagy ensures cellular survival by removing cellular waste and pathogens via the autophagosome-lysosome pathway.

Purpose of the Study:

  • To discuss the application of high content imaging (HCI) and analysis in autophagy research.
  • To highlight the use of HCI in monitoring autophagosomal markers and cargo recruitment.
  • To provide insights into studying autophagosome formation using advanced imaging techniques.

Main Methods:

  • Utilizing fluorescence microscopy to visualize and track autophagosomal markers and cargo.
  • Employing high content imaging (HCI) for automated, quantitative analysis of cellular processes.
  • Analyzing spatial and temporal recruitment of components to the autophagosome.

Main Results:

  • HCI enables efficient and detailed monitoring of autophagosome biogenesis.
  • The study reviews commonly used markers for assessing autophagosomal formation.
  • Quantitative analysis of autophagic flux and marker localization is facilitated by HCI.

Conclusions:

  • High content imaging is a powerful tool for advancing the study of autophagy.
  • HCI facilitates a deeper understanding of the mechanisms underlying autophagosome formation.
  • This approach aids in dissecting the role of autophagy in cellular survival and disease.