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Related Concept Videos

Super-resolution Fluorescence Microscopy01:37

Super-resolution Fluorescence Microscopy

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Related Experiment Video

Updated: Aug 6, 2025

Multimodal Analysis of Microplastics in Drinking Water using a Silicon Nanomembrane Analysis Pipeline
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Published on: June 13, 2025

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Modification of fluorescence staining method for small-sized microplastic quantification: Focus on the interference

Wentao Hu1,2, Rui Tang3,4, Shoujun Yuan5,6

  • 1School of Civil Engineering, Hefei University of Technology, Hefei, 230009, People's Republic of China.

Environmental Science and Pollution Research International
|March 14, 2023
PubMed
Summary

Accurate microplastic quantification using fluorescence staining is challenged by background contamination and procedural losses. This study identified key interference factors and developed effective measures to improve accuracy, aligning results with micro-Raman spectroscopy.

Keywords:
Background contaminationFluorescence staining methodMicroplasticModificationQuantification

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Area of Science:

  • Environmental Science
  • Analytical Chemistry
  • Polymer Science

Background:

  • Microplastics are a pervasive global pollutant, with fluorescence staining being a common quantification method.
  • Existing fluorescence staining methods for microplastics suffer from significant interference, impacting accuracy.
  • Quality assurance measures for microplastic quantification via fluorescence staining are not well-established.

Purpose of the Study:

  • To identify key factors interfering with accurate microplastic quantification using fluorescence staining.
  • To develop and validate effective quality assurance measures to mitigate these interferences.
  • To improve the reliability of fluorescence staining methods for microplastic analysis.

Main Methods:

  • Investigated the Nile Red/DAPI co-staining method as a model system.
  • Assessed background microplastic contamination in lab water, filter membranes, and glassware.
  • Evaluated the impact of hydrogen peroxide (H2O2) pretreatment and exposure time on microplastic assessment.
  • Quantified microplastic loss during the extraction process.
  • Compared fluorescence staining results with micro-Raman spectroscopy.

Main Results:

  • High background microplastic contamination (up to 1115 MP/L) was found in lab waters, filter membranes, and glassware.
  • Simple filtration removed 99% of background microplastics from water; burning eliminated contamination from membranes and glassware.
  • H2O2 pretreatment (48 h) and exposure time (10 ms) were optimized to avoid erroneous size assessment.
  • Significant microplastic loss occurred during extraction (20% for 5 μm, >50% for 20 μm).
  • Modified measures reduced detected microplastic concentrations by 65.7–92.2%, yielding results comparable to micro-Raman spectroscopy.

Conclusions:

  • Background contamination and procedural losses are critical factors affecting fluorescence-based microplastic quantification.
  • Implemented quality assurance measures significantly enhance the accuracy of microplastic detection.
  • This study provides a framework for improving the reliability of fluorescence staining techniques for microplastic analysis.