High-throughput, low-loss, low-cost, and label-free cell separation using electrophysiology-activated cell enrichment.



Faraghat, SA, Hoettges, KF, Steinbach, MK, van der Veen, DR, Brackenbury, WJ, Henslee, EA, Labeed, FH and Hughes, MP
(2017) High-throughput, low-loss, low-cost, and label-free cell separation using electrophysiology-activated cell enrichment. Proceedings of the National Academy of Sciences of the United States of America, 114 (18). 4591 - 4596.

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Abstract

Currently, cell separation occurs almost exclusively by density gradient methods and by fluorescence- and magnetic-activated cell sorting (FACS/MACS). These variously suffer from lack of specificity, high cell loss, use of labels, and high capital/operating cost. We present a dielectrophoresis (DEP)-based cell-separation method, using 3D electrodes on a low-cost disposable chip; one cell type is allowed to pass through the chip whereas the other is retained and subsequently recovered. The method advances usability and throughput of DEP separation by orders of magnitude in throughput, efficiency, purity, recovery (cells arriving in the correct output fraction), cell losses (those which are unaccounted for at the end of the separation), and cost. The system was evaluated using three example separations: live and dead yeast; human cancer cells/red blood cells; and rodent fibroblasts/red blood cells. A single-pass protocol can enrich cells with cell recovery of up to 91.3% at over 300,000 cells per second with >3% cell loss. A two-pass protocol can process 300,000,000 cells in under 30 min, with cell recovery of up to 96.4% and cell losses below 5%, an effective processing rate >160,000 cells per second. A three-step protocol is shown to be effective for removal of 99.1% of RBCs spiked with 1% cancer cells while maintaining a processing rate of ∼170,000 cells per second. Furthermore, the self-contained and low-cost nature of the separator device means that it has potential application in low-contamination applications such as cell therapies, where good manufacturing practice compatibility is of paramount importance.

Item Type: Article
Depositing User: Symplectic Admin
Date Deposited: 21 Apr 2017 08:49
Last Modified: 01 Oct 2021 07:36
DOI: 10.1073/pnas.1700773114
Related URLs:
URI: https://livrepository.liverpool.ac.uk/id/eprint/3007035