Issue 2, 2016

Microfluidic device for simultaneous analysis of neutrophil extracellular traps and production of reactive oxygen species

Abstract

Neutrophil extracellular traps (NETs) were first reported in 2004, and since their discovery, there has been an increasing interest in NETs, how they are formed, their role in controlling infections, and their contribution to disease pathogenesis. Despite this rapid expansion of our understanding of NETs, many details remain unclear including the role of reactive oxygen species (ROS) in the formation of NETs. Further, to study NETs, investigators typically require a large number of cells purified via a lengthy purification regimen. Here, we report a microfluidic device used to quantify both ROS and NET production over time in response to various stimulants, including live bacteria. This device enables ROS and NET analysis using a process that purifies primary human neutrophils in less than 10 minutes and requires only a few microliters of whole blood. Using this device we demonstrate the ability to identify distinct capabilities of neutrophil subsets (including ROS production and NET formation), the ability to use different stimulants/inhibitors, and the ability to effectively use samples stored for up to 8 hours. This device permits the study of ROS and NETs in a user-friendly format and has potential for widespread applications in the study of human disease.

Graphical abstract: Microfluidic device for simultaneous analysis of neutrophil extracellular traps and production of reactive oxygen species

Supplementary files

Article information

Article type
Technical Innovation
Submitted
05 Sep 2015
Accepted
24 Dec 2015
First published
25 Jan 2016

Integr. Biol., 2016,8, 243-252

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