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Local Bayesian Equivalent-Circuit Inversion for Live Dielectric Phenotyping of A549, 293T, and HL-60 Cells

Shen Xi1, Fei Yu1, Wu Yuhan1
1Shanxi Key Lab for Modernization of TCVM, College of Life Science, Shanxi Agricultural University, Taiyuan 030000, Shanxi, P. R. China

Abstract

Label-free impedance flow cytometry is capable of registering the passage of single cells at high speed, but the biological information extracted from the measurement depends on how accurately the transient amplitude-phase pulses are converted to intrinsic electrical variables. This work considers a local Bayesian equivalent-circuit inversion applied to dual-frequency constriction-channel measurements of A549, 293T, and HL-60 cells. The stream of the measurements consists of 100,902 A549/293T events, which have been acquired during ten five-minute acquisition cycles, as well as HL-60 control/treatment sequence, which has been acquired after cytochalasin B and N-formyl-Met-Leu-Phe stimulation. Every measurement is characterized by pre-event and peak amplitude and phase at 100 kHz and 180 kHz. The local Bayesian inversion evaluates the equivalent-circuit mismatch for the neighborhood, which is provided by the current event, fits the local surface of the mismatch, and returns specific membrane capacitance (\(\Csm\)), cytoplasm conductivity (\(\sigcyto\)), and event-level reliability. The A549 events have a peak at \(2.6~\mu\mathrm{F}/\mathrm{cm}^{2}\) and \(0.3~\mathrm{S}/\mathrm{m}\), and the 293T events have a peak at \(1.2~\mu\mathrm{F}/\mathrm{cm}^{2}\) and \(0.4~\mathrm{S}/\mathrm{m}\); same-line Kullback–Leibler divergence between A549/293T distributions equals 0.020–0.060, while A549–293T divergence is equal to 1.824. The exhaustive physical fitting requires \(16800\pm455\) ms per cell, whereas the event-local approach avoids the repetition of the full library and retains the physical uncertainty channel. Treatment of HL-60 cells leads to the shift of the distribution towards low capacitance and increase of Q3 occupancy from 32.35% to 41.51%. The analysis suggests that real-time dielectric phenotyping should not return \(\Csm\) and \(\sigcyto\) as isolated values. The streaming cytometer is reliable if the intrinsic properties are returned together with their event-level evidences, especially in case of distribution movement induced by treatment.

Keywords: impedance flow cytometry, Bayesian inversion, equivalent circuit, single-cell dielectric phenotype, A549 cells, 293T cells, HL-60 cells, uncertainty quantification, constriction microchannel, real-time cytometry
Citation
Shen Xi, Fei Yu, Wu Yuhan. Local Bayesian Equivalent-Circuit Inversion for Live Dielectric Phenotyping of A549, 293T, and HL-60 Cells[J], TK Techforum Journal (ThyssenKrupp Techforum), Volume 2023 (3). 53-71.

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