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Updated: Jul 5, 2026
11:39
Assessing Autophagic Flux by Measuring LC3, p62, and LAMP1 Co-localization Using Multispectral Imaging Flow Cytometry
Published on: July 21, 2017
Methods for assessing autophagy and autophagic cell death
Ezgi Tasdemir1, Lorenzo Galluzzi, M Chiara Maiuri
1INSERM, Unit Apoptosis, Cancer and Immunity, Villejuif, France.
Methods in Molecular Biology (Clifton, N.J.)
|April 22, 2008
Summary
Autophagic cell death involves massive autophagosome accumulation. This study reviews methods to promote, inhibit, and detect autophagy, distinguishing cell death with autophagy from cell death through autophagy.
Area of Science:
- Cell Biology
- Molecular Biology
- Biochemistry
Background:
- Autophagic cell death (type 2) is defined by extensive autophagosome accumulation, distinct from apoptosis (type 1).
- Understanding the mechanisms and detection of autophagy is crucial for distinguishing its role in cell death.
Purpose of the Study:
- To critically assess methods for promoting and inhibiting autophagy.
- To review techniques for detecting autophagy and its role in cell death.
- To provide a strategy for differentiating cell death with autophagy from cell death through autophagy.
Main Methods:
- Pharmacological and genetic manipulations to modulate autophagy.
- Transmission electron microscopy for ultrastructural analysis.
- Biochemical assays including LC3 maturation and protein half-life assessments.
- Fluorescence microscopy for colocalization studies of organelle markers with lysosomes.
Main Results:
- Detailed assessment of various methods to induce and suppress autophagy.
- Comprehensive review of established and emerging techniques for autophagy detection.
- Proposed strategy to discriminate between cell death occurring with autophagy and cell death mediated by autophagy.
Conclusions:
- Autophagy can be pharmacologically and genetically manipulated.
- Accurate detection methods are essential for studying autophagic cell death.
- Distinguishing the cause of cell death in relation to autophagy is critical for biological interpretation.
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