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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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Updated: Jun 6, 2026

Cell-based Flow Cytometry Assay to Measure Cytotoxic Activity

Cell-based Flow Cytometry Assay to Measure Cytotoxic Activity

Published on: December 17, 2013

Tracking immune cell proliferation and cytotoxic potential using flow cytometry.

Joseph D Tario¹, Katharine A Muirhead, Dalin Pan

¹Department of Flow and Image Cytometry, Roswell Park Cancer Institute, Buffalo, NY, USA.

Methods in Molecular Biology (Clifton, N.J.)

|December 1, 2010

Summary

This study details using two cell tracking dyes with flow cytometry to assess cytotoxic effector and regulatory T-cell functions. It provides protocols for analyzing cell viability, proliferation, and cytotoxicity in immune responses.

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Last Updated: Jun 6, 2026

Cell-based Flow Cytometry Assay to Measure Cytotoxic Activity

Cell-based Flow Cytometry Assay to Measure Cytotoxic Activity

Published on: December 17, 2013

Optimized Staining and Proliferation Modeling Methods for Cell Division Monitoring using Cell Tracking Dyes

Optimized Staining and Proliferation Modeling Methods for Cell Division Monitoring using Cell Tracking Dyes

Published on: December 13, 2012

Flow Cytometry-based Assay for the Monitoring of NK Cell Functions

Flow Cytometry-based Assay for the Monitoring of NK Cell Functions

Published on: October 30, 2016

Area of Science:

Immunology
Cell Biology
Flow Cytometry

Background:

Cell tracking dyes and flow cytometry are crucial for analyzing immune cell functions.
Previous work focused on single dye tracking for CD8(+) T cell proliferation and cytokine production.
Assessing complex immune cell functions requires advanced multi-dye methodologies.

Purpose of the Study:

To extend the application of cell tracking dyes to evaluate cytotoxic effector and regulatory T-cell functions.
To provide guidance on combining two different tracking dyes with phenotypic and viability probes.
To detail protocols for two-color cell tracking in immunological assays.

Main Methods:

Utilized two distinct cell tracking dyes (protein- and membrane-labeling) in conjunction with phenotypic and viability probes.
Determined optimal staining concentrations for various dye types.
Developed detailed labeling protocols for dual-dye cell tracking.
Applied two-color cell tracking to cytotoxicity assays and regulatory T-cell function monitoring.

Main Results:

Established protocols for independent enumeration of viable effector and target cells in cytotoxicity assays.
Demonstrated simultaneous monitoring of proliferative responses in effector and regulatory T cells.
Highlighted the advantages of two-color cell tracking for complex immune cell analysis.

Conclusions:

Two-color cell tracking significantly enhances the ability to assess complex immune cell functions, including cytotoxicity and regulatory activity.
The presented protocols offer a robust framework for researchers studying cellular immune responses.
Optimized dye selection and staining protocols are critical for accurate multi-parametric flow cytometry analysis.