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Updated: Sep 12, 2025

Discrimination of Seven Immune Cell Subsets by Two-fluorochrome Flow Cytometry
Published on: March 5, 2019
Advanced immunophenotyping of lymphocyte and monocyte subsets in healthy Australian adults using a novel spectral
Ainsley R Davies1,2, Kristy Kwong1,2,3, Zhijia Yu1
1Canberra Clinical Phenomics Service, John Curtin School of Medical Research, The Australian National University, Canberra, ACT, Australia.
Introduction:
Lymphocytes play pivotal roles in disease pathogenesis and can be used as potential biomarkers for various immunological conditions. Yet, current flow cytometry methods used in clinical settings are often only capable of measuring between four to eight distinct lymphocyte populations. The purpose of our study was to measure many lymphocyte and monocyte populations from a single sample, with the long-term aim of validating our assay for diagnostic use in the Australian regulatory environment.
Methods:
We designed and optimised a novel 30-colour lymphocyte immunophenotyping panel tailored for use on a 3-laser (V-B-R) spectral flow cytometer. This panel measures over 50 lymphocyte and monocyte populations.
Results:
In this report we present data derived from 148 healthy individuals.
Discussion:
This lays the groundwork for future clinical application of spectral flow cytometry tests and offers a more comprehensive approach to lymphocyte and monocyte analysis with future implications for disease diagnosis and monitoring.

