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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: Sep 13, 2025

Isolation of CD4+ T-cells and Analysis of Circulating T-follicular Helper cTfh Cell Subsets from Peripheral Blood Using 6-color Flow Cytometry

Isolation of CD4+ T-cells and Analysis of Circulating T-follicular Helper cTfh Cell Subsets from Peripheral Blood Using 6-color Flow Cytometry

Published on: January 7, 2019

Efficient Tonsillar T Follicular Helper Cell Processing and Functional Analysis through High-dimensional Flow

Qin Xu¹, Lihong Shi¹, Brian L P Dizon²

¹Cell Signaling and Immunity Section, Laboratory of Immune System Biology, National Institute of Allergy and Infectious Diseases, National Institutes of Health.

Journal of Visualized Experiments : Jove

|July 28, 2025

Summary

This summary is machine-generated.

This study details a protocol for human tonsil cell isolation and culture, crucial for studying T follicular helper (Tfh) cells. It also presents flow cytometry methods and data analysis techniques for Tfh cell research.

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Last Updated: Sep 13, 2025

Isolation of CD4+ T-cells and Analysis of Circulating T-follicular Helper cTfh Cell Subsets from Peripheral Blood Using 6-color Flow Cytometry

Isolation of CD4+ T-cells and Analysis of Circulating T-follicular Helper cTfh Cell Subsets from Peripheral Blood Using 6-color Flow Cytometry

Published on: January 7, 2019

High-Dimensionality Flow Cytometry for Immune Function Analysis of Dissected Implant Tissues

High-Dimensionality Flow Cytometry for Immune Function Analysis of Dissected Implant Tissues

Published on: September 15, 2021

Flow Cytometric Characterization of Murine B Cell Development

Flow Cytometric Characterization of Murine B Cell Development

Published on: January 22, 2021

Area of Science:

Immunology
Cell Biology

Background:

T follicular helper (Tfh) cells are critical for adaptive humoral immunity, influencing B cell responses in infection, autoimmunity, and vaccination.
Understanding Tfh cell development and function is key for vaccine design and targeted therapies for immune-related diseases.

Purpose of the Study:

To provide a comprehensive protocol for the collection, isolation, preservation, and culture of human tonsil cells.
To introduce optimized spectral flow cytometry panels for detailed Tfh cell characterization.
To demonstrate the utility of high-dimensional data analysis for uncovering treatment-induced immune cell differences.

Main Methods:

Human tonsil tissue collection, cell isolation, and in vitro culture.
Development and optimization of two spectral flow cytometry panels for Tfh cell phenotyping.
Application of unsupervised analysis on high-dimensional flow cytometry data.

Main Results:

A standardized protocol for human tonsil cell processing was established.
Optimized flow cytometry panels enable in-depth characterization of Tfh cells and related immune populations.
Unsupervised analysis successfully identified subtle, treatment-induced changes in high-dimensional immune cell data.

Conclusions:

Human tonsil cells provide a valuable model for studying Tfh cell biology and immune responses.
The presented protocol and flow cytometry panels facilitate robust Tfh cell research.
High-dimensional data analysis is a powerful tool for discovering immune cell alterations in response to stimuli or treatments.