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Integrated inertial-impedance cytometry for rapid label-free leukocyte isolation and profiling of neutrophil extracellular traps (NETs)

Abstract

Circulating leukocytes are indispensible components of the immune system, and rapid analysis of their native state or functionalities can help to unravel their pathophysiological roles and identify novel prognostics biomarkers in health and diseases. Herein we report a novel high throughput “sample-in-answer-out” integrated platform for continuous leukocyte sorting and single-cell electrical profiling in a label-free manner. The multi-staged platform enables isolation of neutrophils and monocytes from diluted or lysed blood samples directly within minutes based on Dean Flow Fractionation (DFF) (stage 1). Next DFF-purified leukocytes are inertially focused in serpertine channels into single stream (stage 2) prior impedance detection (stage 3). As a proof-of-concept for neutrophil functional characterization towards diabetes testing, we characterised the formation of neutrophil extracellular traps (NETosis) of healthy and glucose-treated neutrophils and observed significant changes in dielectric properties (size and opacity) between both groups. Interestingly, the NETosis profile induced by calcium ionophore (CaI) and phorbol 12-myristate 13-acetate (PMA) were also electrically different, which could be attributed to the differential rates of cell enlargement and attentuated membrane permeability. Taken together, these results clearly demonstrated the potential of the developed platform for rapid (~mins) and label-free leukocyte profiling, and the use of impedance signatures as novel functional biomarkers for point-of-care testing in diabetes.

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Publication details

The article was received on 14 Mar 2019, accepted on 12 Apr 2019 and first published on 15 Apr 2019


Article type: Paper
DOI: 10.1039/C9LC00250B
Citation: Lab Chip, 2019, Accepted Manuscript

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    Integrated inertial-impedance cytometry for rapid label-free leukocyte isolation and profiling of neutrophil extracellular traps (NETs)

    C. Petchakup, H. M. Tay, K. H. H. Li and H. W. Hou, Lab Chip, 2019, Accepted Manuscript , DOI: 10.1039/C9LC00250B

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