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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: Mar 30, 2026

Author Spotlight: Unveiling the Polyfunctionality and Heterogeneity in Immune Responses

Author Spotlight: Unveiling the Polyfunctionality and Heterogeneity in Immune Responses

Published on: March 8, 2024

Microfluidic Flow Cytometry for Single-Cell Protein Analysis.

Meiye Wu¹, Anup K Singh^2,3

¹Biotechnology and Bioengineering, Sandia National Laboratories, Livermore, CA, USA.

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

|November 7, 2015

Summary

This summary is machine-generated.

Microfluidic flow cytometry (FC) enables rapid, automated single-cell protein analysis. This method profiles cell surface and intracellular proteins, demonstrating TLR4 activation and cytokine production in macrophages.

Keywords:

Cell surface receptors Cytokine Flow cytometry Immunostaining Microfluidic Phosphoprotein Single-cell resolution

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Last Updated: Mar 30, 2026

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Area of Science:

Biotechnology
Cell Biology
Analytical Chemistry

Background:

Single-cell protein analysis is crucial for understanding cellular heterogeneity.
Traditional flow cytometry (FC) has limitations in automation and throughput for complex analyses.
Microfluidic technologies offer miniaturization and automation for advanced cell analysis.

Purpose of the Study:

To describe a method for microfluidic flow cytometry (FC).
To provide instructions for building a microfluidic platform for automated cell analysis.
To demonstrate the platform's capability in profiling specific cellular responses.

Main Methods:

Development of a microfluidic platform for automated cell culture and immunostaining.
Integration of microfluidic chip with micro-flow cytometry for analysis.
Immunostaining for cell surface receptors, intracellular phosphoproteins, and cytokines.
Profiling of Toll-like receptor 4 (TLR4) activation, ERK1/2 phosphorylation, and TNFα production.

Main Results:

Successful implementation of automated cell culture and immunostaining on the microfluidic platform.
Quantitative analysis of cell surface and intracellular protein markers in single cells.
Demonstration of TLR4 receptor activation, ERK1/2 phosphorylation, and TNFα production in LPS-stimulated macrophages.
Validation of the microfluidic FC platform for complex cellular pathway analysis.

Conclusions:

Microfluidic FC provides a powerful, automated solution for single-cell protein analysis.
The developed platform enables comprehensive profiling of cellular responses, including receptor activation and cytokine production.
This technology facilitates high-throughput investigation of cellular signaling pathways and biomarker discovery.