Quantification of capture efficiency, purity, and single-cell isolation in the recovery of circulating melanoma cells from peripheral blood by dielectrophoresis

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2023-02-07
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Chen, Han
Osman, Sommer Y.
Moose, Devon L.
Vanneste, Marion
Henry, Michael D.
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ChemRxiv
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Anand, Robbyn
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Chemistry

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This paper describes a dielectrophoretic method for selection of circulating melanoma cells (CMCs), which lack reliable identifying surface antigens and are extremely rare in blood. This platform captures CMCs individually by dielectrophoresis (DEP) at an array of wireless bipolar electrodes (BPEs) aligned to overlying nanoliter-scale chambers, which isolate each cell for subsequent on-chip single-cell analysis. To determine the best conditions to employ for CMC isolation in this DEP-BPE platform, the static and dynamic dielectrophoretic response of established melanoma cell lines, melanoma cells from patient-derived xenografts (PDX) and peripheral blood mononuclear cells (PBMCs) were evaluated as a function of frequency using two established DEP platforms. Further, PBMCs derived from patients with advanced melanoma were compared with those from healthy controls. The results of this evaluation reveal that each DEP method requires a distinct frequency to achieve capture of melanoma cells and that the distribution of dielectric properties of PBMCs is more broadly varied in and among patients versus healthy controls. Based on this evaluation, we conclude that 50 kHz provides the highest capture efficiency on our DEP-BPE platform while maintaining a low rate of capture of unwanted PBMCs. We further quantified the efficiency of single-cell capture on the DEP-BPE platform and found that the efficiency diminished beyond around 25% chamber occupancy, thereby informing the minimum array size that is required. Importantly, the capture efficiency of the DEP-BPE platform for melanoma cells when using optimized conditions matched the performance predicted by our analysis. Finally, isolation of melanoma cells from contrived (spike-in) and clinical samples on our platform using optimized conditions was demonstrated. The capture and individual isolation of CMCs, confirmed by post-capture labeling, from patient-derived samples suggests the potential of this platform for clinical application.
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This is a pre-print of the article Chen, Han, Sommer Osman, Devon Moose, Marion Vanneste, Jared Anderson, Michael Henry, and Robbyn Anand. “Quantification of Capture Efficiency, Purity, and Single-Cell Isolation in the Recovery of Circulating Melanoma Cells from Peripheral Blood by Dielectrophoresis.” ChemRxiv, 2023. DOI: 10.26434/chemrxiv-2023-cs884. Attribution-NonCommercial 4.0 International (CC BY-NC 4.0). Copyright 2023 The Authors. Posted with permission.
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