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Related Experiment Video

Updated: Mar 6, 2026

Quantitative Analysis of Autophagy using Advanced 3D Fluorescence Microscopy

Quantitative Analysis of Autophagy using Advanced 3D Fluorescence Microscopy

Published on: May 3, 2013

Quantifying Autophagic Structures in Mammalian Cells Using Confocal Microscopy.

C A Lamb¹, J Joachim¹, S A Tooze¹

¹Molecular Cell Biology of Autophagy Group, The Francis Crick Institute, London, United Kingdom.

Methods in Enzymology

|March 4, 2017

Summary

This summary is machine-generated.

This study details methods for observing autophagy progression by tracking autophagosome formation and the movement of ATG9 protein. These techniques aid in understanding cellular waste removal processes under starvation.

Keywords:

ATG9 Autophagosome Autophagy Imaris LC3 Live cell imaging

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Published on: July 21, 2017

Live Cell Imaging of Early Autophagy Events: Omegasomes and Beyond

Live Cell Imaging of Early Autophagy Events: Omegasomes and Beyond

Published on: July 27, 2013

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Last Updated: Mar 6, 2026

Quantitative Analysis of Autophagy using Advanced 3D Fluorescence Microscopy

Quantitative Analysis of Autophagy using Advanced 3D Fluorescence Microscopy

Published on: May 3, 2013

Assessing Autophagic Flux by Measuring LC3, p62, and LAMP1 Co-localization Using Multispectral Imaging Flow Cytometry

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Published on: July 21, 2017

Live Cell Imaging of Early Autophagy Events: Omegasomes and Beyond

Live Cell Imaging of Early Autophagy Events: Omegasomes and Beyond

Published on: July 27, 2013

Area of Science:

Cell Biology
Molecular Biology

Background:

Autophagy is a fundamental cellular process for degrading and recycling damaged components.
The formation of autophagosomes and the trafficking of autophagy-related proteins, like ATG9, are crucial for efficient autophagy.
Monitoring these dynamic events is essential for understanding cellular health and disease.

Purpose of the Study:

To describe methodologies for quantifying autophagosome numbers in mammalian cells.
To present techniques for tracking the intracellular trafficking of the ATG9 protein.
To provide tools for investigating autophagy dynamics under specific cellular conditions, such as starvation.

Main Methods:

Confocal microscopy is utilized to visualize and quantify autophagosome formation.
Autophagosomal markers are employed to track the sequential assembly of phagophores into autophagosomes.
Live-cell imaging techniques are used to monitor the dynamic movement of ATG9 in response to starvation.

Main Results:

The described methods allow for precise measurement of autophagosome biogenesis and maturation.
The trafficking patterns of ATG9 under starvation conditions can be effectively visualized and analyzed.
These techniques provide quantitative data on key steps in the autophagy pathway.

Conclusions:

The presented methods offer robust approaches for studying autophagy dynamics at the molecular and cellular levels.
Understanding ATG9 trafficking and autophagosome formation is key to deciphering autophagy regulation.
These techniques can be applied to further research into autophagy-related disorders.