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Related Concept Videos

Flow Cytometry01:23

Flow Cytometry

The development of flow cytometry techniques began in 1934 with initial attempts by Andrew Moldavan, a bacteriologist who counted the cells in a flowing capillary system. Moldavan pumped cells through a capillary tube focused under a microscope for visualization. The invention of photometry allowed the measurement of differentially-stained cells, and Louis Kamentsky developed the first multiparameter flow cytometer in 1965 to identify and count the cancer cells in cervical tissue specimens.
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Related Experiment Video

Updated: Jun 13, 2026

Assessing Autophagic Flux by Measuring LC3, p62, and LAMP1 Co-localization Using Multispectral Imaging Flow Cytometry
11:39

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

Published on: July 21, 2017

A novel quantitative flow cytometry-based assay for autophagy.

Kai Er Eng1, Marc D Panas, Gunilla B Karlsson Hedestam

  • 1Department of Microbiology, Tumor and Cell Biology, Karolinska Institutet, Stockholm, Sweden.

Autophagy
|May 12, 2010
PubMed
Summary
This summary is machine-generated.

We developed a simple flow cytometry assay to measure autophagy, a key cellular process. This method quantifies autophagosome levels in both transfected and non-transfected cells, aiding research into nutrient sensing and stress responses.

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Quantitative Analysis of Autophagy using Advanced 3D Fluorescence Microscopy
09:59

Quantitative Analysis of Autophagy using Advanced 3D Fluorescence Microscopy

Published on: May 3, 2013

Related Experiment Videos

Last Updated: Jun 13, 2026

Assessing Autophagic Flux by Measuring LC3, p62, and LAMP1 Co-localization Using Multispectral Imaging Flow Cytometry
11:39

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

Published on: July 21, 2017

Quantitative Analysis of Autophagy using Advanced 3D Fluorescence Microscopy
09:59

Quantitative Analysis of Autophagy using Advanced 3D Fluorescence Microscopy

Published on: May 3, 2013

Area of Science:

  • Cell Biology
  • Molecular Biology

Background:

  • Autophagy is a fundamental cellular degradation process vital for nutrient sensing, stress responses, and development.
  • Understanding autophagy flux is crucial for deciphering its role in various biological pathways.

Purpose of the Study:

  • To present a straightforward and quantitative assay for measuring autophagy.
  • To adapt the assay for differentiating constitutive and induced autophagy and quantifying flux.
  • To enable detection of autophagosome formation in non-transfected cells.

Main Methods:

  • Utilized a EGFP-LC3 reporter protein combined with flow cytometry.
  • Employed saponin extraction specific for EGFP-LC3-I to quantify autophagosome-associated EGFP-LC3-II.
  • Incorporated degradation inhibitors to analyze autophagy flux.
  • Developed direct antibody staining for endogenous LC3 detection.

Main Results:

  • The assay accurately measures autophagosome-associated EGFP-LC3-II levels via flow cytometry.
  • The method successfully differentiates and quantifies changes in constitutive and induced autophagy flux.
  • Autophagosome formation can be detected in non-transfected cells using antibody staining.

Conclusions:

  • This flow cytometry-based assay provides a sensitive, high-throughput, and quantitative method for studying autophagy.
  • The assay is versatile, applicable to both transfected and non-transfected cells, and allows for detailed flux analysis.
  • This tool will advance research into the diverse biological roles of autophagy.