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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 12, 2026

Analyzing Cellular Internalization of Nanoparticles and Bacteria by Multi-spectral Imaging Flow Cytometry

Analyzing Cellular Internalization of Nanoparticles and Bacteria by Multi-spectral Imaging Flow Cytometry

Published on: June 8, 2012

An improved method for differentiating cell-bound from internalized particles by imaging flow cytometry.

Asya Smirnov¹, Michael D Solga¹, Joanne Lannigan¹

¹Department of Microbiology, Immunology, and Cancer Biology, University of Virginia, Charlottesville, VA, USA.

Journal of Immunological Methods

|May 14, 2015

Summary

This study introduces a novel imaging flow cytometry protocol to quantify particle binding and internalization by cells. The method accurately distinguishes extracellular from intracellular particles, advancing phagocytosis research.

Keywords:

Attachment Bacteria Imaging flow cytometry Internalization Phagocytosis

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Last Updated: Apr 12, 2026

Analyzing Cellular Internalization of Nanoparticles and Bacteria by Multi-spectral Imaging Flow Cytometry

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Published on: June 8, 2012

Image-based Flow Cytometry Technique to Evaluate Changes in Granulocyte Function In Vitro

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Quantification of Efferocytosis by Single-cell Fluorescence Microscopy

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Published on: August 18, 2018

Area of Science:

Cell Biology
Immunology
Biotechnology

Background:

Phagocytosis is crucial for pathogen clearance and cellular homeostasis.
Existing methods lack high-throughput quantification and discrimination of bound versus internalized particles.

Purpose of the Study:

To develop and validate a rapid, high-throughput protocol for quantifying particle association and internalization by cells.
To enable discrimination between extracellularly bound and intracellularly phagocytosed particles.

Main Methods:

Utilized imaging flow cytometry to analyze cells exposed to fluorescent particles.
Applied a novel antibody staining technique to differentiate extracellular particles.
Employed a spot count algorithm to quantify particle numbers per cell.

Main Results:

The protocol successfully quantified attached and internalized fluorescent particles in cell populations.
Validated the spot count algorithm using fluorescence and phase contrast imaging.
Demonstrated the method's efficacy in studying Neisseria gonorrhoeae internalization by neutrophils.

Conclusions:

The developed imaging flow cytometry protocol offers a powerful and rapid method for assessing particle uptake.
This technique facilitates detailed analysis of phagocytosis dynamics across various cell types and conditions.