Document Type

Article

Department

​Chemistry​

Publication Date

6-2018

Abstract

Microfluidic chemical cytometry is a powerful technique for examining chemical contents of individual cells, but applications have focused on cells from multicellular organisms, especially mammals. We demonstrate the first use of microfluidic chemical cytometry to examine a unicellular organism, the social amoeba Dictyostelium discoideum. We used the reactive oxygen species indicator dichlorodihydrofluorescein diacetate to report on oxidative stress and controlled for variations in indicator loading and retention using carboxyfluorescein diacetate as an internal standard. After optimizing indicator concentration, we investigated the effect of peroxide treatment through single-cell measurements of 353 individual cells. The peak area ratio of dichlorofluorescein to carboxyfluorescein increased from 1.69 ± 0.89 for untreated cells to 5.19 ± 2.72 for cells treated with 40 mM hydrogen peroxide. Interestingly, the variance of the data also increased with oxidative stress. While preliminary, these results are consistent with the hypothesis that heterogeneous stress responses in unicellular organisms may be adaptive.

Comments

Author's accepted manuscript provided by Trinity College Digital Repository in accordance with publisher's distribution policies.

Kathy Rodogiannis and Jessica Duong both Trinity College students.

Published version available:

Kathy Rodogiannis, Jessica T. Duong, and Michelle L. Kovarik. “Microfluidic Single-cell Analysis of Oxidative Stress in Dictyostelium discoideum.” Analyst 143 (2018): 3643-3650. DOI: 10.1039/C8AN00752G
https://doi.org/10.1039/C8AN00752G

Publication Title

Analyst

Volume

143

DOI

10.1039/C8AN00752G

Available for download on Saturday, June 01, 2019

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