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Open AccessFeature PaperArticle

Single Cell Analysis of Neutrophils NETs by Microscopic LSPR Imaging System

1
Department of Applied Physics, Graduate School of Engineering, Osaka University, 2-1 Yamadaoka, Suita 565-0871, Japan
2
Department of Electric and Electronic Engineering, Universiti Tun Hussein Onn Malaysia, Parit Raja, Batu Pahat 86400, Johor, Malaysia
3
Advanced Photonics and Biosensing Open Innovation Laboratory, AIST–Osaka University, Photonic Center Osaka University, Suita, Osaka 565-0871, Japan
4
Department of Respiratory Medicine and Clinical Immunology, Graduate School of Medicine, Osaka University, 2-2 Yamadaoka, Suita, Osaka 565-0871, Japan
*
Author to whom correspondence should be addressed.
Micromachines 2020, 11(1), 52; https://doi.org/10.3390/mi11010052
Received: 9 December 2019 / Revised: 27 December 2019 / Accepted: 29 December 2019 / Published: 31 December 2019
(This article belongs to the Special Issue Micro and Nano Devices for Cell Analysis)
A simple microengraving cell monitoring method for neutrophil extracellular traps (NETs) released from single neutrophils has been realized using a polydimethylsiloxane (PDMS) microwell array (MWA) sheet on a plasmon chip platform. An imbalance between NETs formation and the succeeding degradation (NETosis) are considered associated with autoimmune disease and its pathogenesis. Thus, an alternative platform that can conduct monitoring of this activity on single cell level at minimum cost but with great sensitivity is greatly desired. The developed MWA plasmon chips allow single cell isolation of neutrophils from 150 µL suspension (6.0 × 105 cells/mL) with an efficiency of 36.3%; 105 microwells with single cell condition. To demonstrate the utility of the chip, trapped cells were incubated between 2 to 4 h after introducing with 100 nM phorbol 12-myristate 13-acetate (PMA) before measurement. Under observation using a hyperspectral imaging system that allows high-throughput screening, the neutrophils stimulated by PMA solution show a significant release of fibrils and NETs after 4 h, with observed maximum areas between 314–758 µm2. An average absorption peak wavelength shows a redshift of Δλ = 1.5 nm as neutrophils release NETs.
Keywords: neutrophil; localized surface plasmon resonance (LSPR); microwell neutrophil; localized surface plasmon resonance (LSPR); microwell
MDPI and ACS Style

Ahmad Mohamed Ali, R.; Mita, D.; Espulgar, W.; Saito, M.; Nishide, M.; Takamatsu, H.; Yoshikawa, H.; Tamiya, E. Single Cell Analysis of Neutrophils NETs by Microscopic LSPR Imaging System. Micromachines 2020, 11, 52.

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