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Modification of a Nile Red Staining Method for Microplastics Analysis: A Nile Red Plate Method

Kang, Heejun, Park, Saerom, Lee, Bokjin, Ahn, Jaehwan, Kim, Seogku·2020·Water
DOI 10.3390/w12113251
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Abstract

Recently, environmental pollution from microplastics (MPs) has become a significant reason for increasing the number of studies to develop analysis methods. The Nile red staining method (NR-S), which is staining polymer particles with Nile red (NR) dye, has been widely used for the analysis of MPs in environmental samples. However, this method has several limitations, as it is difficult to stain MPs covered with organic matter residues. In this study, we modified the NR-S method into an NR plate method (NR-P), where the plate is coated with NR instead of staining MPs directly. The optimum concentration of NR solution was obtained (1000 mg/L), and the effectiveness of the NR-P method for the analysis of MPs was assessed using different types (polypropylene, polyethylene, polyethylene terephthalate, and polystyrene), sizes (100–1000 µm), and shapes (sphere, fiber, film, and flake) of plastic materials. The NR-P method demonstrated improved resolution in the overall types, shapes, and sizes of MPs and was better than the control (without NR plate method) and NR-S method. In particular, the NR-P method can effectively observe MPs covered with organic matter, which was a major limitation of the NR-S method. Finally, MPs in sewage field samples were analyzed by the NR-P method with an accuracy of 78% confirmed by FT-IR. We demonstrated that this method is a convenient and efficient alternative for identifying MPs, even for field samples.

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Figure 1 ( a ) Nile red plate analysis with respect to solution concentration and ( b ) identification of polypropylene (PP) particle under a microscope ( 30).
Figure 1 ( a ) Nile red plate analysis with respect to solution concentration and ( b ) identification of polypropylene (PP) particle under a microscope ( 30).CC-BY · From the PDF
Image 2
CC-BY · From the PDF
Figure 2 Identification of plastic types sized in the range of 300 to 500 µ m (PP, polyethylene (PE), polyethylene terephthalate (PET), and polystyrene (PS)).
Figure 2 Identification of plastic types sized in the range of 300 to 500 µ m (PP, polyethylene (PE), polyethylene terephthalate (PET), and polystyrene (PS)).CC-BY · From the PDF
Figure 3 Identification of microplastics based on size. ( a ) Above 1000 µ m, ( b ) 700~1000 µ m, ( c ) 400~700 µ m, and ( d ) 100-400 µ m).
Figure 3 Identification of microplastics based on size. ( a ) Above 1000 µ m, ( b ) 700~1000 µ m, ( c ) 400~700 µ m, and ( d ) 100-400 µ m).CC-BY · From the PDF
Figure 4 Identification of shape of microplastics on the Nile red plate (NR-P) and Nile red staining (NR-S) (PS: sphere, Polyester (PEs): fiber, PE: film, and PP: flake) and recovery rate.
Figure 4 Identification of shape of microplastics on the Nile red plate (NR-P) and Nile red staining (NR-S) (PS: sphere, Polyester (PEs): fiber, PE: film, and PP: flake) and recovery rate.CC-BY · From the PDF
Figure 5 Identification of e ect of organic matter contamination by control and NR-P methods.
Figure 5 Identification of e ect of organic matter contamination by control and NR-P methods.CC-BY · From the PDF
Figure 6 Identification of field samples: ( a ) sludge and ( b ) sewage influent under ( left ) control and ( right ) NR-P methods ( 30).
Figure 6 Identification of field samples: ( a ) sludge and ( b ) sewage influent under ( left ) control and ( right ) NR-P methods ( 30).CC-BY · From the PDF
Figure 7 Results of FT-IR analysis of luminescent particles in the field samples using the NR-P method.
Figure 7 Results of FT-IR analysis of luminescent particles in the field samples using the NR-P method.CC-BY · From the PDF

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Metadata

Authors
Kang, H., Park, S., Lee, B., Ahn, J., & Kim, S.
Venue
Water
Publisher
MDPI AG
Published
2020-11-19
Volume
12
Issue
11
Pages
3251
ISSN
2073-4441
DOI
10.3390/w12113251
Cited by
58
References
3
Language
en
Licence
creativecommons.org/licenses/by/4.0/ (vor)
Funders
  • Korea institute of Civil Engineering and Building Technology(KICT) (20200394-001)
  • National Research Council of Science &amp; Technology (NST) (CAP-18-07-KICT)

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CoChat. (2026). Citation check: Modification of a Nile Red Staining Method for Microplastics Analysis: A Nile Red Plate Method. https://app.cochat.ai/papers/10.3390/w12113251
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Kang, H., Park, S., Lee, B., Ahn, J., & Kim, S. (2020). Modification of a Nile Red Staining Method for Microplastics Analysis: A Nile Red Plate Method. Water. https://doi.org/10.3390/w12113251