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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: Apr 26, 2026

Flow Cytometric Characterization of Murine B Cell Development

Flow Cytometric Characterization of Murine B Cell Development

Published on: January 22, 2021

Classification of common variable immunodeficiencies using flow cytometry and a memory B-cell functionality assay.

Amelia L Rösel¹, Carmen Scheibenbogen², Ulrike Schliesser³

¹Institute for Medical Immunology, Charité University Medicine Berlin, Berlin, Germany.

The Journal of Allergy and Clinical Immunology

|August 13, 2014

Summary

A new assay reveals memory B cell (memBc) functionality in common variable immunodeficiency (CVID), suggesting potential for new therapies. This functional assessment could lead to improved CVID classification and patient care.

Keywords:

Memory B cell antibody-secreting cells common variable immunodeficiency subtyping enzyme-linked immunosorbent spot assay flow cytometry

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Area of Science:

Immunology
Cellular Biology
Clinical Medicine

Background:

Common variable immunodeficiency (CVID) is a heterogeneous disorder characterized by hypogammaglobulinemia, recurrent infections, autoimmunity, and malignancy.
Memory B cells (memBcs) are crucial for humoral immunity, but their functional capacity is not currently used for CVID classification.

Purpose of the Study:

To establish and validate a memory B cell enzyme-linked immunosorbent spot (ELISpot) assay to assess B cell function in CVID.
To propose a novel CVID classification system based on memBc functional capacity.

Main Methods:

Application of a memBc ELISpot assay combined with flow cytometry in CVID patients and age-matched healthy controls.
Analysis of ex vivo and in vitro antibody secretion and proliferation of memBcs.

Main Results:

Significantly reduced ex vivo frequencies of IgG-, IgM-, and IgA-secreting plasmablasts in CVID patients compared to controls.
Impaired in vitro differentiation of memBcs into antibody-secreting cells across all immunoglobulin isotypes.
Inverse correlation between memBc proliferation and immunoglobulin secretion, suggesting compensatory hyperproliferation.
Identification of heterogeneous memBc function within CVID patients, enabling subclassification.

Conclusions:

The developed memBc ELISpot assay effectively characterizes B cell functional capacity in CVID.
Functional assessment of memBcs may allow for a more precise CVID subclassification.
Restored memBc/plasmablast functionality under specific stimulation suggests potential for novel therapeutic strategies in CVID.