MethodsX (Jan 2023)

Easy and accessible way to calibrate a fluorescence microscope and to create a microplastic identification key

  • Anna Kukkola,
  • Stefan Krause,
  • Yasmin Yonan,
  • Liam Kelleher,
  • Uwe Schneidewind,
  • Gregory H. Sambrook Smith,
  • Holly Nel,
  • Iseult Lynch

Journal volume & issue
Vol. 10
p. 102053

Abstract

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We present here a technique for setting up detection limits on any fluorescent microscope in conjunction with the fluorophore Nile Red for microplastic identification. Our method also describes a rigorous morphology-specific identification key for microplastics to reduce subjectivity between researchers. The detection limits were established for nine common polymer types and five natural substrates which could result in false-positive signals when using Nile Red for microplastic identification. This method was then applied to real freshwater samples and identified particles were validated with micro-FTIR or Raman spectroscopy. This approach may reduce subjectivity in microplastic identification and counting and enhances transparency, repeatability and harmonization within microplastic research community. • Instructions for calibration of detection limits for microplastics on fluorescence microscope systems described • Microplastic identification key developed and tested to reduce false positive detection • Lower subjectivity for microplastic identification obtained using the detection limits & identification key

Keywords