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Related Concept Videos

Autophagy01:27

Autophagy

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Autophagy is a self-digesting process by which a cell protects itself from threats both within and outside the cell, ranging from abnormal proteins to invading bacteria. In this process, obsolete components of the cell and invading microbes are degraded by hydrolytic enzymes active in an acidic environment of the lysosomal lumen.
An autophagic pathway consists of a series of signaling events activated in response to diverse stress and physiological conditions such as food deprivation,...
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Related Experiment Video

Updated: Oct 29, 2025

Assessing Autophagic Flux by Measuring LC3, p62, and LAMP1 Co-localization Using Multispectral Imaging Flow Cytometry
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Autophagy assessment in circulating leukocytes.

José Manuel Bravo-San Pedro1, Federico Pietrocola2

  • 1Universidad Complutense de Madrid, Departamento de Fisiología, Facultad de Medicina, Madrid, Spain.

Methods in Cell Biology
|July 6, 2021
PubMed
Summary

Researchers developed a simple, non-invasive method to measure autophagy in mice. This "autophagometer" uses circulating leukocytes to validate pro-autophagy drug actions, aiding healthspan and lifespan research.

Keywords:
AgingBloodCaloric restrictionFastingLC3LeupeptinLysosomeMetabolism

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Area of Science:

  • Cellular Biology
  • Biochemistry
  • Aging Research

Background:

  • Autophagy is a key regulator of healthspan and lifespan.
  • There is a need for simple, non-invasive methods to measure autophagy in vivo.
  • Fasting is a known activator of autophagy.

Purpose of the Study:

  • To develop and validate a novel method for quantifying autophagic flux in circulating leukocytes.
  • To establish a preclinical tool for assessing pro-autophagy interventions.

Main Methods:

  • A leupeptin-based assay was employed.
  • Video-flow cytometry was used for detection of LC3B puncta.
  • The method was applied to circulating leukocytes in mice.

Main Results:

  • The developed system reliably quantifies autophagy in mouse leukocytes.
  • The assay demonstrated sensitivity to fasting-induced autophagy.
  • The method serves as a potential "autophagometer" for preclinical studies.

Conclusions:

  • A novel, non-invasive method to measure autophagic flux in vivo has been established.
  • This technique can be routinely used in experimental settings.
  • The method is suitable for validating the efficacy of pro-autophagy drugs.