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A Simple Micromilled Microfluidic Impedance Cytometer with Vertical Parallel Electrodes for Cell Viability Analysis
Eades, Jason Audiffred, Julianne F. Fincher, Micah Choi, Jin-Woo Soper, Steven A. Monroe, William Todd
Eades, Jason
Audiffred, Julianne F.
Fincher, Micah
Choi, Jin-Woo
Soper, Steven A.
Monroe, William Todd
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Abstract
Microfluidic impedance cytometry has been demonstrated as an effective platform for single cell analysis, taking advantage of microfabricated features and dielectric cell sensing methods. In this study, we present a simple microfluidic device to improve the sensitivity, accuracy, and throughput of single suspension cell viability analysis using vertical sidewall electrodes fabricated by a widely accessible negative manufacturing method. A microchannel milled through a 75 µm platinum wire, which was embedded into poly-methyl-methacrylate (PMMA), created a pair of parallel vertical sidewall platinum electrodes. Jurkat cells were interrogated in a custom low-conductivity buffer (1.2 ± 0.04 mS/cm) to reduce current leakage and increase device sensitivity. Confirmed by live/dead staining and electron microscopy, a single optimum excitation frequency of 2 MHz was identified at which live and dead cells were discriminated based on the disruption in the cell membrane associated with cell death. At this frequency, live cells were found to exhibit changes in the impedance phase with no appreciable change in magnitude, while dead cells displayed the opposite behavior. Correlated with video microscopy, a computational algorithm was created that could identify cell detection events and determine cell viability status by application of a mathematical correlation method.
Description
Date
2023-01-22
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Publisher
MDPI
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Keywords
Single cell, Microfluidic impedance cytometry, Impedance spectroscopy, Micromilling, Vertical electrodes, Sidewall electrodes
Citation
Eades, J.; Audiffred, J.F.; Fincher, M.; Choi, J.-W.; Soper, S.A.; Monroe, W.T. A Simple Micromilled Microfluidic Impedance Cytometer with Vertical Parallel Electrodes for Cell Viability Analysis. Micromachines 2023, 14, 283. https://doi.org/10.3390/mi14020283