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Updated: Sep 11, 2025

Discrimination of Seven Immune Cell Subsets by Two-fluorochrome Flow Cytometry
Published on: March 5, 2019
Fourier goniometric flow cytometry modeled to show improved particle differentiation beyond forward and side scatter
Abstract:
Flow Cytometry (FC) has proven to be an indispensable means of differentiating between sample populations in both clinical and general biological applications. Traditional FC and fluorescence FC integrate optical signals to measure total intensities among the forward scattering (FSC), side scattering (SSC), and fluorescent channels to differentiate between populations. This paper introduces Fourier Goniometric Flow Cytometry (FGFC) as a means of augmenting traditional FC by adding angular resolution to FSC and SSC channels. The FGFC system is simulated and a novel, to the best of our knowledge, single-valued Fourier Goniometric Flow Cytometry (svFGFC) metric is introduced. Results are analyzed using AUC analysis of ROC curves for traditional FC and FGFC. Traditional FC versus FGFC was simulated 500 times for populations with nuanced scattering differences, and FGFC consistently outperformed traditional FC across all simulations. The svFGFC metric consistently shows an improvement over traditional FC scattering measurements in differentiating between sample populations with subtle differences, showing an average improvement of 4% over the AUC of traditional FC. The added resolution afforded by FGFC promises to improve differentiation between sample populations with subtle differences based upon scattering alone. Moreover, the addition of a more efficient, purely scatter-based metric for differentiating particle classification in FC could help to relieve the increasing dependence upon fluorescence FC. FGFC would, at the least, further complement fluorescence FC.

