Single cell analysis of neutrophils NETs by Microscopic LSPR imaging system

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 degradatio...

Full description

Bibliographic Details
Main Authors: Mohamed Ali, Riyaz Ahmad, Mita, Daiki, Espulgar, Wilfred, Saito, Masato, Nishide, Masayuki, Takamatsu, Hyota, Yoshikawa, Hiroyuki, Tamiya, Eiichi
Format: Article
Language:English
Published: MDPI 2020
Subjects:
Online Access:http://eprints.uthm.edu.my/605/
http://eprints.uthm.edu.my/605/1/DNJ9662_14ea9c16ee4b846e6b6bf660dae04c2b.pdf
_version_ 1848887241700540416
author Mohamed Ali, Riyaz Ahmad
Mita, Daiki
Espulgar, Wilfred
Saito, Masato
Nishide, Masayuki
Takamatsu, Hyota
Yoshikawa, Hiroyuki
Tamiya, Eiichi
author_facet Mohamed Ali, Riyaz Ahmad
Mita, Daiki
Espulgar, Wilfred
Saito, Masato
Nishide, Masayuki
Takamatsu, Hyota
Yoshikawa, Hiroyuki
Tamiya, Eiichi
author_sort Mohamed Ali, Riyaz Ahmad
building UTHM Institutional Repository
collection Online Access
description 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.
first_indexed 2025-11-15T19:51:16Z
format Article
id uthm-605
institution Universiti Tun Hussein Onn Malaysia
institution_category Local University
language English
last_indexed 2025-11-15T19:51:16Z
publishDate 2020
publisher MDPI
recordtype eprints
repository_type Digital Repository
spelling uthm-6052021-08-11T03:49:53Z http://eprints.uthm.edu.my/605/ Single cell analysis of neutrophils NETs by Microscopic LSPR imaging system Mohamed Ali, Riyaz Ahmad Mita, Daiki Espulgar, Wilfred Saito, Masato Nishide, Masayuki Takamatsu, Hyota Yoshikawa, Hiroyuki Tamiya, Eiichi TA1501-1820 Applied optics. Photonics 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. MDPI 2020 Article PeerReviewed text en http://eprints.uthm.edu.my/605/1/DNJ9662_14ea9c16ee4b846e6b6bf660dae04c2b.pdf Mohamed Ali, Riyaz Ahmad and Mita, Daiki and Espulgar, Wilfred and Saito, Masato and Nishide, Masayuki and Takamatsu, Hyota and Yoshikawa, Hiroyuki and Tamiya, Eiichi (2020) Single cell analysis of neutrophils NETs by Microscopic LSPR imaging system. Micromachines, 11 (52). pp. 1-15. https://doi.org/10.3390/mi11010052
spellingShingle TA1501-1820 Applied optics. Photonics
Mohamed Ali, Riyaz Ahmad
Mita, Daiki
Espulgar, Wilfred
Saito, Masato
Nishide, Masayuki
Takamatsu, Hyota
Yoshikawa, Hiroyuki
Tamiya, Eiichi
Single cell analysis of neutrophils NETs by Microscopic LSPR imaging system
title Single cell analysis of neutrophils NETs by Microscopic LSPR imaging system
title_full Single cell analysis of neutrophils NETs by Microscopic LSPR imaging system
title_fullStr Single cell analysis of neutrophils NETs by Microscopic LSPR imaging system
title_full_unstemmed Single cell analysis of neutrophils NETs by Microscopic LSPR imaging system
title_short Single cell analysis of neutrophils NETs by Microscopic LSPR imaging system
title_sort single cell analysis of neutrophils nets by microscopic lspr imaging system
topic TA1501-1820 Applied optics. Photonics
url http://eprints.uthm.edu.my/605/
http://eprints.uthm.edu.my/605/
http://eprints.uthm.edu.my/605/1/DNJ9662_14ea9c16ee4b846e6b6bf660dae04c2b.pdf