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Towards quality-assured measurements of microplastics in soil using fluorescence microscopy

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Towards quality-assured measurements of microplastics in soil using fluorescence microscopy. / Le, Quynh Nhu Phan; Halsall, Crispin; Peneva, Stoyana et al.
In: Analytical and Bioanalytical Chemistry, Vol. 417, No. 11, 31.05.2025, p. 2225-2238.

Research output: Contribution to Journal/MagazineJournal articlepeer-review

Harvard

Le, QNP, Halsall, C, Peneva, S, Wrigley, O, Braun, M, Amelung, W, Ashton, L, Surridge, BWJ & Quinton, J 2025, 'Towards quality-assured measurements of microplastics in soil using fluorescence microscopy', Analytical and Bioanalytical Chemistry, vol. 417, no. 11, pp. 2225-2238. https://doi.org/10.1007/s00216-025-05810-6

APA

Le, Q. N. P., Halsall, C., Peneva, S., Wrigley, O., Braun, M., Amelung, W., Ashton, L., Surridge, B. W. J., & Quinton, J. (2025). Towards quality-assured measurements of microplastics in soil using fluorescence microscopy. Analytical and Bioanalytical Chemistry, 417(11), 2225-2238. Advance online publication. https://doi.org/10.1007/s00216-025-05810-6

Vancouver

Le QNP, Halsall C, Peneva S, Wrigley O, Braun M, Amelung W et al. Towards quality-assured measurements of microplastics in soil using fluorescence microscopy. Analytical and Bioanalytical Chemistry. 2025 May 31;417(11):2225-2238. Epub 2025 Mar 10. doi: 10.1007/s00216-025-05810-6

Author

Le, Quynh Nhu Phan ; Halsall, Crispin ; Peneva, Stoyana et al. / Towards quality-assured measurements of microplastics in soil using fluorescence microscopy. In: Analytical and Bioanalytical Chemistry. 2025 ; Vol. 417, No. 11. pp. 2225-2238.

Bibtex

@article{3f00d582be2043d8ab829907898616c1,
title = "Towards quality-assured measurements of microplastics in soil using fluorescence microscopy",
abstract = "Fluorescence microscopy is increasingly seen as a fast, user-friendly, and high-throughput method for detecting microplastics (MPs) in soil; however, its effectiveness across diverse MP types and soil properties remains underexplored. This study tested a fluorescence microscopy-Nile red (NR) staining approach on eight MP types, covering both biodegradable and non-biodegradable plastics, in three size ranges (≤ 150 µm, 100-250 µm, 500-1000 µm) across loamy, clayey, and sandy soils. Each sample, processed in triplicate, underwent a relatively quick and straightforward extraction procedure involving density separation, organic digestion, and NR staining, followed by fluorescence and bright-field microscopy. A new digital image analysis pipeline using Image J was developed to expedite and (semi)automate MP quantification. Recoveries ranged from 80% to 90% for MPs with a Feret diameter of 500-1000 µm, regardless of soil type. In contrast, the recovery of smaller MPs (Feret dia. ≤ 250 µm) varied depending on the soils and plastic types: recoveries for low-density polyethylene (LDPE) reached 85% in sandy soil and 90% in loamy soil, whereas those for biodegradable polybutylene adipate terephthalate/polylactic acid (PBAT/PLA) were only 60% and 10%, respectively. The lowest recovery rate was observed in clayey soil and for biodegradable plastics. The method was tested on non-agricultural soil samples, yielding a MP mean number concentration of 20.7 ± 9.0 MPs/g for MPs sized from dia. ≥ 25 µm, comparable to Fourier transform infrared (FPA-µ-FTIR) results of 13.1 ± 7.3 MPs/g (p > 0.05). We conclude that fluorescence microscopy with NR staining and automated particle quantification offers a time-efficient, reproducible, and accurate method for MP detection in light-textured soils, whereas limitations remain for reliable MP analysis in clay-dominated soils.",
keywords = "Nile red staining, Polymers, Fluorescence microscopy, Soil organic matter",
author = "Le, {Quynh Nhu Phan} and Crispin Halsall and Stoyana Peneva and Olivia Wrigley and Melanie Braun and Wulf Amelung and Lorna Ashton and Surridge, {Ben W J} and John Quinton",
year = "2025",
month = mar,
day = "10",
doi = "10.1007/s00216-025-05810-6",
language = "English",
volume = "417",
pages = "2225--2238",
journal = "Analytical and Bioanalytical Chemistry",
issn = "1618-2642",
publisher = "Springer Verlag",
number = "11",

}

RIS

TY - JOUR

T1 - Towards quality-assured measurements of microplastics in soil using fluorescence microscopy

AU - Le, Quynh Nhu Phan

AU - Halsall, Crispin

AU - Peneva, Stoyana

AU - Wrigley, Olivia

AU - Braun, Melanie

AU - Amelung, Wulf

AU - Ashton, Lorna

AU - Surridge, Ben W J

AU - Quinton, John

PY - 2025/3/10

Y1 - 2025/3/10

N2 - Fluorescence microscopy is increasingly seen as a fast, user-friendly, and high-throughput method for detecting microplastics (MPs) in soil; however, its effectiveness across diverse MP types and soil properties remains underexplored. This study tested a fluorescence microscopy-Nile red (NR) staining approach on eight MP types, covering both biodegradable and non-biodegradable plastics, in three size ranges (≤ 150 µm, 100-250 µm, 500-1000 µm) across loamy, clayey, and sandy soils. Each sample, processed in triplicate, underwent a relatively quick and straightforward extraction procedure involving density separation, organic digestion, and NR staining, followed by fluorescence and bright-field microscopy. A new digital image analysis pipeline using Image J was developed to expedite and (semi)automate MP quantification. Recoveries ranged from 80% to 90% for MPs with a Feret diameter of 500-1000 µm, regardless of soil type. In contrast, the recovery of smaller MPs (Feret dia. ≤ 250 µm) varied depending on the soils and plastic types: recoveries for low-density polyethylene (LDPE) reached 85% in sandy soil and 90% in loamy soil, whereas those for biodegradable polybutylene adipate terephthalate/polylactic acid (PBAT/PLA) were only 60% and 10%, respectively. The lowest recovery rate was observed in clayey soil and for biodegradable plastics. The method was tested on non-agricultural soil samples, yielding a MP mean number concentration of 20.7 ± 9.0 MPs/g for MPs sized from dia. ≥ 25 µm, comparable to Fourier transform infrared (FPA-µ-FTIR) results of 13.1 ± 7.3 MPs/g (p > 0.05). We conclude that fluorescence microscopy with NR staining and automated particle quantification offers a time-efficient, reproducible, and accurate method for MP detection in light-textured soils, whereas limitations remain for reliable MP analysis in clay-dominated soils.

AB - Fluorescence microscopy is increasingly seen as a fast, user-friendly, and high-throughput method for detecting microplastics (MPs) in soil; however, its effectiveness across diverse MP types and soil properties remains underexplored. This study tested a fluorescence microscopy-Nile red (NR) staining approach on eight MP types, covering both biodegradable and non-biodegradable plastics, in three size ranges (≤ 150 µm, 100-250 µm, 500-1000 µm) across loamy, clayey, and sandy soils. Each sample, processed in triplicate, underwent a relatively quick and straightforward extraction procedure involving density separation, organic digestion, and NR staining, followed by fluorescence and bright-field microscopy. A new digital image analysis pipeline using Image J was developed to expedite and (semi)automate MP quantification. Recoveries ranged from 80% to 90% for MPs with a Feret diameter of 500-1000 µm, regardless of soil type. In contrast, the recovery of smaller MPs (Feret dia. ≤ 250 µm) varied depending on the soils and plastic types: recoveries for low-density polyethylene (LDPE) reached 85% in sandy soil and 90% in loamy soil, whereas those for biodegradable polybutylene adipate terephthalate/polylactic acid (PBAT/PLA) were only 60% and 10%, respectively. The lowest recovery rate was observed in clayey soil and for biodegradable plastics. The method was tested on non-agricultural soil samples, yielding a MP mean number concentration of 20.7 ± 9.0 MPs/g for MPs sized from dia. ≥ 25 µm, comparable to Fourier transform infrared (FPA-µ-FTIR) results of 13.1 ± 7.3 MPs/g (p > 0.05). We conclude that fluorescence microscopy with NR staining and automated particle quantification offers a time-efficient, reproducible, and accurate method for MP detection in light-textured soils, whereas limitations remain for reliable MP analysis in clay-dominated soils.

KW - Nile red staining

KW - Polymers

KW - Fluorescence microscopy

KW - Soil organic matter

U2 - 10.1007/s00216-025-05810-6

DO - 10.1007/s00216-025-05810-6

M3 - Journal article

C2 - 40063098

VL - 417

SP - 2225

EP - 2238

JO - Analytical and Bioanalytical Chemistry

JF - Analytical and Bioanalytical Chemistry

SN - 1618-2642

IS - 11

ER -