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Flow Cytometry

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: Apr 16, 2026

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

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

Published on: July 21, 2017

Quantifying autophagosomes and autolysosomes in cells using imaging flow cytometry.

Robin Rajan¹, Magdalena Karbowniczek, Haley R Pugsley

¹School of Pharmacy, Department of Immunotherapeutics and Biotechnology, Texas Tech University Health Sciences Center, Abilene, Texas.

Cytometry. Part a : the Journal of the International Society for Analytical Cytology

|March 3, 2015

Summary

This study introduces a new method to accurately measure autophagy by combining Bright Detail Similarity (BDS) with microtubule associated protein 1B light chain 3 (LC3) puncta counts. This approach enhances the quantification of autophagosomes and autolysosomes in disease research.

Keywords:

LAMP1 LC3 autolysosome autophagosome autophagy imaging flow cytometry spot count

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Last Updated: Apr 16, 2026

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Area of Science:

Cell Biology
Molecular Biology
Biochemistry

Background:

Autophagy dysregulation is linked to various diseases.
Accurate monitoring of autophagy is crucial for disease pathogenesis and therapeutic development.
Existing methods like Bright Detail Similarity (BDS) for imaging flow cytometry have limitations in quantifying autophagic flux.

Purpose of the Study:

To develop an improved method for quantifying autophagic processing.
To address the limitations of BDS in accounting for the number of autophagic organelles.
To enhance the understanding of autophagy in disease and treatment.

Main Methods:

Developed a novel imaging flow cytometry approach combining Bright Detail Similarity (BDS) with the count of microtubule associated protein 1B light chain 3 (LC3) puncta.
Applied the method to breast cancer cells under basal conditions, chloroquine-induced inhibition, and amino acid starvation-induced autophagy.
Quantified autophagosomes and autolysosomes to differentiate between cells with few organelles versus those with accumulated autophagosomes/autolysosomes.

Main Results:

The combined BDS and LC3 puncta counting method effectively distinguished cells with varying autophagosome accumulation.
The new approach provided a more nuanced evaluation of autophagic processing compared to BDS alone.
Successfully quantified autophagosomes and autolysosomes in breast cancer cells under different experimental conditions.

Conclusions:

The developed method offers a more comprehensive assessment of autophagy by integrating organelle number and lysosomal colocalization.
This technique provides valuable insights into autophagy processing, serving as a useful supplement to existing methods.
Enhanced autophagy monitoring can significantly aid in understanding disease mechanisms and evaluating therapeutic strategies.