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

doi:10.1016/j.mex.2023.102053

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

^*Corresponding author for this work

Research output: Contribution to journal › Article › peer-review

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Abstract

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

Original language	English
Article number	102053
Number of pages	8
Journal	MethodsX
Volume	10
Early online date	3 Feb 2023
DOIs	https://doi.org/10.1016/j.mex.2023.102053
Publication status	E-pub ahead of print - 3 Feb 2023

Keywords

Microplastics
Identification key
Nile Red
Fluorescence microscopy
Particle counting
Fibres
Fragments
Detection
Detection limits
Identification

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10.1016/j.mex.2023.102053Licence: Creative Commons: Attribution (CC BY)

KukkolaA2023EasyFinal published version, 1.41 MBLicence: Creative Commons: Attribution (CC BY)

International Freshwater Microplastics Network
Sambrook-Smith, G. (Principal Investigator), Krause, S. (Co-Investigator) & Lynch, I. (Co-Investigator)
Natural Environment Research Council
18/12/19 → 17/12/21
Project: Research Councils
Plastic Rivers- Fate and transport of microplastics in freshwater riverine environments
Lynch, I. (Co-Investigator), Krause, S. (Principal Investigator) & Sambrook-Smith, G. (Co-Investigator)
Leverhulme Trust
1/08/18 → 31/03/23
Project: Research

Cite this

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title = "Easy and accessible way to calibrate a fluorescence microscope and to create a microplastic identification key",

abstract = "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 = "Microplastics, Identification key, Nile Red, Fluorescence microscopy, Particle counting, Fibres, Fragments, Detection, Detection limits, Identification",

author = "Anna Kukkola and Stefan Krause and Yasmin Yonan and Liam Kelleher and Uwe Schneidewind and Smith, {Gregory H Sambrook} and Holly Nel and Iseult Lynch",

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AU - Kukkola, Anna

AU - Krause, Stefan

AU - Yonan, Yasmin

AU - Kelleher, Liam

AU - Schneidewind, Uwe

AU - Smith, Gregory H Sambrook

AU - Nel, Holly

AU - Lynch, Iseult

PY - 2023/2/3

Y1 - 2023/2/3

N2 - 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

AB - 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

KW - Microplastics

KW - Identification key

KW - Nile Red

KW - Fluorescence microscopy

KW - Particle counting

KW - Fibres

KW - Fragments

KW - Detection

KW - Detection limits

KW - Identification

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DO - 10.1016/j.mex.2023.102053

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VL - 10

JO - MethodsX

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

Abstract

Keywords

Access to Document

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Projects

International Freshwater Microplastics Network

Plastic Rivers- Fate and transport of microplastics in freshwater riverine environments

Cite this