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Updated: Aug 8, 2026
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Preparation of Whole Bone Marrow for Mass Cytometry Analysis of Neutrophil-lineage Cells
Published on: June 19, 2019
Human peripheral B cells: a different cytometric point of view
Maurizio Carbonari1, Tiziana Tedesco, Massimo Fiorilli
1Department of Clinical Medicine, University of Rome La Sapienza, Viale dell'Università 37, 00185 Rome, Italy. maurizio.carbonari@uniroma1.it
Summary
This study introduces apparent fluorescence density analysis for human B lymphocytes, correlating fluorescence intensity with cell size. This method reveals unique surface molecule expression patterns in B cell disorders, improving flow cytometry analysis.
Area of Science:
- Immunology
- Cell Biology
- Biotechnology
Background:
- Human peripheral B lymphocytes exhibit complex flow cytometry signals.
- Current methods often report fluorescence intensity without considering cell surface area.
- A novel approach using fluorescence intensity to forward scatter ratio is proposed.
Purpose of the Study:
- To introduce a new method for analyzing B lymphocyte surface molecules.
- To determine the apparent fluorescence density of surface molecules.
- To improve the interpretation of flow cytometry data.
Main Methods:
- Analysis of list mode data from live B cells.
- Logical gating of B cell populations.
- Serial scanning of FSC vs. SSC plots to calculate apparent fluorescence density.
Main Results:
- Detected steady and modulated fluorescence densities for various surface molecules on normal B lymphocytes.
- Observed distinct alterations in phenotype density values and distributions in B cells from patients with disorders.
- Demonstrated the ability to differentiate size-dependent fluorescence variations.
Conclusions:
- Apparent fluorescence density analysis in human B cell cytometry accounts for cell size variations.
- This method uncovers expression patterns not detectable by conventional intensity-based approaches.
- Preliminary data suggest clinical utility in diagnosing B cell disorders.
