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Exploring the Efficacy of Nile Red in Microplastic Quantification: A Costaining Approach

Stanton, Thomas; Johnson, Matthew; Nathanail, Paul; Gomes, Rachel; Needham, Teresa; Burson, Amanda


Thomas Stanton

Paul Nathanail

Professor of Water & Resource Processing

Teresa Needham

Amanda Burson


The presence of microplastic particles ([less than]5 mm) in the environment has generated considerable concern across public, political, and scientific platforms. However, the diversity of microplastics that persist in the environment poses complex analytical challenges for our understanding of their prevalence. The use of the dye Nile red to quantify microplastics is increasingly common. However, its use in microplastic analysis rarely accounts for its affinity with the breadth of particles that occur in environmental samples. Here, we examine Nile red’s ability to stain a variety of microplastic particles and common natural and anthropogenic particles found in environmental samples. To better constrain microplastic estimates using Nile red, we test the coapplication of a second stain that binds to biological material, 4′,6-diamidino-2-phenylindole (DAPI). We test the potential inflation of microplastic estimates using Nile red alone by applying this costaining approach to samples of drinking water and freshwater. The use of Nile red dye alone resulted in a maximum 100% overestimation of microplastic particles. These findings are of particular significance for the public dissemination of findings from an emotive field of study.


Stanton, T., Johnson, M., Nathanail, P., Gomes, R., Needham, T., & Burson, A. (2019). Exploring the Efficacy of Nile Red in Microplastic Quantification: A Costaining Approach. Environmental Science and Technology Letters, 6(10), 606-611.

Journal Article Type Article
Acceptance Date Sep 19, 2019
Online Publication Date Sep 19, 2019
Publication Date Oct 8, 2019
Deposit Date Sep 26, 2019
Publicly Available Date Sep 20, 2020
Journal Environmental Science & Technology Letters
Electronic ISSN 2328-8930
Publisher American Chemical Society
Peer Reviewed Peer Reviewed
Volume 6
Issue 10
Pages 606-611
Public URL
Publisher URL
Additional Information This document is the Accepted Manuscript version of a Published Work that appeared in final form in Environmental Science and Technology Letters, copyright © American Chemical Society after peer review and technical editing by the publisher. To access the final edited and published work see


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