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Detection limits are central to improve reporting standards when using Nile red for microplastic quantification

Chemosphere 2020 101 citations ? Citation count from OpenAlex, updated daily. May differ slightly from the publisher's own count. Score: 40 ? 0–100 AI score estimating relevance to the microplastics field. Papers below 30 are filtered from public browse.
Simeon Onoja, Simeon Onoja, Stefan Krause, Stefan Krause, Stefan Krause, Stefan Krause, Andrew J. Chetwynd, Liam Kelleher, Liam Kelleher, Liam Kelleher, Holly Nel Liam Kelleher, Henar Margenat, Stefan Krause, Iseult Lynch, Iseult Lynch, Holly Nel Holly Nel Stefan Krause, Holly Nel Henar Margenat, Holly Nel Holly Nel Henar Margenat, Stefan Krause, Stefan Krause, Holly Nel Holly Nel Stefan Krause, Stefan Krause, Henar Margenat, Stefan Krause, Stefan Krause, Henar Margenat, Henar Margenat, Holly Nel Holly Nel Imogen Mansfield, Simeon Onoja, Simeon Onoja, Andrew J. Chetwynd, Stefan Krause, Stefan Krause, Stefan Krause, Stefan Krause, Holly Nel Henar Margenat, Henar Margenat, Henar Margenat, Holly Nel Holly Nel Holly Nel Holly Nel Holly Nel Holly Nel Holly Nel Holly Nel Holly Nel Gregory H. Sambrook Smith, Andrew J. Chetwynd, Andrew J. Chetwynd, Stefan Krause, Liam Kelleher, Iseult Lynch, Iseult Lynch, Iseult Lynch, Iseult Lynch, Iseult Lynch, Iseult Lynch, Stefan Krause, Stefan Krause, Stefan Krause, Stefan Krause, Stefan Krause, Henar Margenat, Holly Nel Holly Nel Henar Margenat, Henar Margenat, Holly Nel Henar Margenat, Holly Nel Henar Margenat, Stefan Krause, Stefan Krause, Stefan Krause, Stefan Krause, Stefan Krause, Stefan Krause, Stefan Krause, Stefan Krause, Stefan Krause, Stefan Krause, Stefan Krause, Stefan Krause, Stefan Krause, Stefan Krause, Iseult Lynch, Iseult Lynch, Iseult Lynch, Iseult Lynch, Iseult Lynch, Iseult Lynch, Iseult Lynch, Iseult Lynch, Iseult Lynch, Iseult Lynch, Iseult Lynch, Liam Kelleher, Liam Kelleher, Liam Kelleher, Gregory H. Sambrook Smith, Stefan Krause, Stefan Krause, Stefan Krause, Stefan Krause, Stefan Krause, Stefan Krause, Stefan Krause, Stefan Krause, Stefan Krause, Iseult Lynch, Iseult Lynch, Gregory H. Sambrook Smith, Iseult Lynch, Iseult Lynch, Iseult Lynch, Gregory H. Sambrook Smith, Henar Margenat, Iseult Lynch, Iseult Lynch, Henar Margenat, Henar Margenat, Holly Nel Henar Margenat, Liam Kelleher, Liam Kelleher, Liam Kelleher, Liam Kelleher, Liam Kelleher, Henar Margenat, Gregory H. Sambrook Smith, Pola Goldberg Oppenheimer, Stefan Krause, Liam Kelleher, Liam Kelleher, Imogen Mansfield, Stefan Krause, Iseult Lynch, Iseult Lynch, Iseult Lynch, Iseult Lynch, Stefan Krause, Holly Nel Iseult Lynch, Stefan Krause, Stefan Krause, Henar Margenat, Henar Margenat, Holly Nel Iseult Lynch, Holly Nel Stefan Krause, Liam Kelleher, Stefan Krause, Stefan Krause, Henar Margenat, Stefan Krause, Holly Nel Holly Nel Liam Kelleher, Iseult Lynch, Iseult Lynch, Liam Kelleher, Pola Goldberg Oppenheimer, Gregory H. Sambrook Smith, Stefan Krause, Stefan Krause, Stefan Krause, Iseult Lynch, Stefan Krause, Iseult Lynch, Gregory H. Sambrook Smith, Holly Nel Stefan Krause, Henar Margenat, Gregory H. Sambrook Smith, Gregory H. Sambrook Smith, Iseult Lynch, Holly Nel Holly Nel Stefan Krause, Stefan Krause, Liam Kelleher, Liam Kelleher, Holly Nel Liam Kelleher, Andrew J. Chetwynd, Stefan Krause, Stefan Krause, Stefan Krause, Gregory H. Sambrook Smith, Stefan Krause, Stefan Krause, Iseult Lynch, Gregory H. Sambrook Smith, Iseult Lynch, Iseult Lynch, Gregory H. Sambrook Smith, Henar Margenat, Iseult Lynch, Liam Kelleher, Iseult Lynch, Stefan Krause, Stefan Krause, Stefan Krause, Iseult Lynch, Stefan Krause, Iseult Lynch, Stefan Krause, Gregory H. Sambrook Smith, Stefan Krause, Iseult Lynch, Iseult Lynch, Iseult Lynch, Simeon Onoja, Holly Nel Iseult Lynch, Holly Nel Stefan Krause, Iseult Lynch, Iseult Lynch, Iseult Lynch, Iseult Lynch, Stefan Krause, Stefan Krause, Iseult Lynch, Iseult Lynch, Stefan Krause, Pola Goldberg Oppenheimer, Iseult Lynch, Stefan Krause, Iseult Lynch, Gregory H. Sambrook Smith, Stefan Krause, Stefan Krause, Stefan Krause, Stefan Krause, Iseult Lynch, Stefan Krause, Stefan Krause, Iseult Lynch, Iseult Lynch, Iseult Lynch, Iseult Lynch, Stefan Krause, Iseult Lynch, Iseult Lynch, Henar Margenat, Gregory H. Sambrook Smith, Iseult Lynch, Iseult Lynch, Henar Margenat, Iseult Lynch, Stefan Krause, Liam Kelleher, Iseult Lynch, Iseult Lynch, Iseult Lynch, Iseult Lynch, Iseult Lynch, Holly Nel Iseult Lynch, Stefan Krause, Iseult Lynch, Holly Nel Holly Nel Holly Nel Iseult Lynch, Holly Nel

Summary

Researchers evaluated the efficacy of Nile red fluorescent staining for quantifying microplastics, demonstrating that polymer-specific differences in fluorescent response create variable detection limits that undermine the comparability and reproducibility of MP quantification across studies. The authors call for standardized reporting of detection limits when using Nile red to improve data quality in microplastic research.

Beyond simple identification of either the presence or absence of microplastic particles in the environment, quantitative accuracy has been criticised as being neither comparable nor reproducible. This is, in part, due to difficulties in the identification of synthetic particles amidst naturally occurring organic and inorganic components. The fluorescent stain Nile red has been proposed as a tool to overcome this issue, but to date, has been used without consideration of polymer specific fluorescent variability. The aim of this study was to evaluate the efficacy of Nile red for microplastic detection by systematically investigating what drives variations in particle pixel brightness (PPB). The results showed that PPB varied between polymer type, shape, size, colour and by staining procedure. Sand, an inorganic component of the sample matrix does not fluoresce when stained with Nile red. In contrast the organic components, wood and chitin, fluoresce between 1.40 and 12 arbitrary units (a.u.) and 32 and 74 a.u. after Nile red staining, respectively. These data informed the use of a PPB threshold limit of 100 a.u., which improved the detection of EPS, HDPE, PP and PA-6 from the 6 polymers tested and reduced analysis time by 30-58% compared to unstained samples. Conversely, as with traditional illumination, PET and PVC were not accurately estimated using this approach. This study shows that picking a threshold limit is not arbitrary but rather must be informed by polymer specific fluorescent variability and matrix considerations. This is an essential step needed to facilitate comparability and reproducibility between individual studies.

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