AAAR 37th Annual Conference October 14 - October 18, 2019 Oregon Convention Center Portland, Oregon, USA
Abstract View
Time-Resolved Single Cell Response of Intracellular Reactive Oxygen Species to Aerosol Particles
FOBANG LIU, Josh Whitley, Nga Lee Ng, Hang Lu, Georgia Institute of Technology
Abstract Number: 336 Working Group: Health-Related Aerosols
Abstract Particulate matter (PM) exposure is a leading global human health risk. Adverse PM health effects have been proposed to be linked to oxidative stress through the generation of reactive oxygen species (ROS). Currently, most assays for cellular response measurement are low resolution (e.g., no single-cell information; not much temporal information), imposing limitations on our understanding of what components or characteristics of PM best account for its toxicity. In this study, we design and optimize microfluidic cell-trapping devices that allow for the long-term culture and imaging of single cells. For method validation, we employ the same bulk intracellular ROS assay (murine alveolar macrophages; fluorescent ROS probe: carboxy-H2DCFDA) that we developed in our previous work. The average cellular ROS response of single-cell measurements is compared with the bulk cellular ROS data obtained via microplate reader. We find that the average data from single cell measurements are higher than that of the bulk data. This could be due to stagnant PM extracts in the well plates having a smaller impact than the continuously perfused PM extracts of the microfluidic device. Furthermore, we link the behaviors of cellular heterogeneity in ROS response to the chemical properties (e.g. oxidation state) of PM. Overall, this study describes a novel approach to understand cellular ROS generation over time at the single-cell level. Our work can serve as a proof of principle study to investigate a wide array of single-cell responses (beyond ROS) upon exposure to any types of PM.