Single Cell Analysis of Neutrophils NETs by Microscopic LSPR Imaging System

Micromachines

Wilfred Espulgar¹  Eiichi Tamiya¹  Daiki Mita¹  Riyaz Ahmad Mohamed Ali¹  Masato Saito¹  Hiroyuki Yoshikawa¹  Hyota Takamatsu²  Masayuki Nishide² 

[1] Department of Applied Physics, Graduate School of Engineering, Osaka University, 2-1 Yamadaoka, Suita 565-0871, Japan;Department of Respiratory Medicine and Clinical Immunology, Graduate School of Medicine, Osaka University, 2-2 Yamadaoka, Suita, Osaka 565-0871, Japan;

关键词: neutrophil;    localized surface plasmon resonance (lspr);    microwell;   

DOI  :  10.3390/mi11010052

来源: DOAJ

【 摘 要 】

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.

【 授权许可】

Unknown