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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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Multiparametric cytometry for exploration of complex cellular dynamics.

Françoise Gondois-Rey¹, Samuel Granjeaud, Suong Le Thi Kieu

¹Institut National de la Santé et de la Recherche Médicale, Unité 891, Centre de Recherche en Cancérologie de Marseille, France. francoise.gondois-rey@inserm.fr

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

|January 27, 2012

Summary

Polychromatic cytometry reveals innate immune responses to influenza virus in human cells. This method tracks activation markers in dendritic cells and NK cells, uncovering complex cellular interactions.

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Area of Science:

Immunology
Virology
Cell Biology

Background:

Polychromatic cytometry has advanced human immunology research.
Previous studies focused on rare cell populations.
This study explores complex innate immune systems using polychromatic cytometry.

Purpose of the Study:

Investigate the dynamics of complex cellular systems in innate immunity.
Utilize polychromatic cytometry to study peripheral blood mononuclear cells (PBMC) stimulated with live influenza virus.
Analyze the time course of PBMC activation, including dendritic cells (DC), monocytes, and NK cells.

Main Methods:

Developed 12-color panels for polychromatic cytometry.
Simultaneously measured intracellular expression of IFN-α, TNF-α, IL-12, IL-6, IFN-γ, CD107, and influenza virus nucleoprotein.
Performed kinetic experiments using low cell numbers and reduced reagents, coupled with bioinformatic tools for data analysis.

Main Results:

Reproducibly determined activation markers in DC and NK cells.
Revealed insights into indirect DC-mediated NK cell activation.
Generated dynamic clustering maps for visualizing PBMC activation events and cellular interactions.

Conclusions:

Polychromatic cytometry is a powerful tool for exploring complex cellular systems in immunology and virology.
The study provides new information on the coordination of innate immune pathways and cellular interactions during viral infection.
This approach enables discoveries in previously unexplored complex cell systems.