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      TUM-ParticleTyper: A detection and quantification tool for automated analysis of (Microplastic) particles and fibers

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          Abstract

          TUM-ParticleTyper is a novel program for the automated detection, quantification and morphological characterization of fragments, including particles and fibers, in images from optical, fluorescence and electron microscopy (SEM). It can be used to automatically select targets for subsequent chemical analysis, e.g., Raman microscopy, or any other single particle identification method. The program was specifically developed and validated for the analysis of microplastic particles on gold coated polycarbonate filters. Our method development was supported by the design of a filter holder that minimizes filter roughness and facilitates enhanced focusing for better images and Raman measurements. The TUM-ParticleTyper software is tunable to the user’s specific sample demands and can extract the morphological characteristics of detected objects (coordinates, Feret’s diameter min / max, area and shape). Results are saved in csv-format and contours of detected objects are displayed as an overlay on the original image. Additionally, the program can stitch a set of images to create a full image out of several smaller ones. An additional useful feature is the inclusion of a statistical process to calculate the minimum number of particles that must be chemically identified to be representative of all particles localized on the substrate. The program performance was evaluated on genuine microplastic samples. The TUM-ParticleTyper software localizes particles using an adaptive threshold with results comparable to the “gold standard” method (manual localization by an expert) and surpasses the commonly used Otsu thresholding by doubling the rate of true positive localizations. This enables the analysis of a statistically significant number of particles on the filter selected by random sampling, measured via single point approach. This extreme reduction in measurement points was validated by comparison to chemical imaging, applying both procedures to the same area at comparable processing times. The single point approach was both faster and more accurate proving the applicability of the presented program.

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          Current research trends on plastic pollution and ecological impacts on the soil ecosystem: A review

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            Analysis of environmental microplastics by vibrational microspectroscopy: FTIR, Raman or both?

            The contamination of aquatic ecosystems with microplastics has recently been reported through many studies, and negative impacts on the aquatic biota have been described. For the chemical identification of microplastics, mainly Fourier transform infrared (FTIR) and Raman spectroscopy are used. But up to now, a critical comparison and validation of both spectroscopic methods with respect to microplastics analysis is missing. To close this knowledge gap, we investigated environmental samples by both Raman and FTIR spectroscopy. Firstly, particles and fibres >500 μm extracted from beach sediment samples were analysed by Raman and FTIR microspectroscopic single measurements. Our results illustrate that both methods are in principle suitable to identify microplastics from the environment. However, in some cases, especially for coloured particles, a combination of both spectroscopic methods is necessary for a complete and reliable characterisation of the chemical composition. Secondly, a marine sample containing particles <400 μm was investigated by Raman imaging and FTIR transmission imaging. The results were compared regarding number, size and type of detectable microplastics as well as spectra quality, measurement time and handling. We show that FTIR imaging leads to significant underestimation (about 35 %) of microplastics compared to Raman imaging, especially in the size range <20 μm. However, the measurement time of Raman imaging is considerably higher compared to FTIR imaging. In summary, we propose a further size division within the smaller microplastics fraction into 500-50 μm (rapid and reliable analysis by FTIR imaging) and into 50-1 μm (detailed and more time-consuming analysis by Raman imaging). Graphical Abstract Marine microplastic sample (fraction <400 μm) on a silicon filter (middle) with the corresponding Raman and IR images.
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              A rapid-screening approach to detect and quantify microplastics based on fluorescent tagging with Nile Red

              A new approach is presented for analysis of microplastics in environmental samples, based on selective fluorescent staining using Nile Red (NR), followed by density-based extraction and filtration. The dye adsorbs onto plastic surfaces and renders them fluorescent when irradiated with blue light. Fluorescence emission is detected using simple photography through an orange filter. Image-analysis allows fluorescent particles to be identified and counted. Magnified images can be recorded and tiled to cover the whole filter area, allowing particles down to a few micrometres to be detected. The solvatochromic nature of Nile Red also offers the possibility of plastic categorisation based on surface polarity characteristics of identified particles. This article details the development of this staining method and its initial cross-validation by comparison with infrared (IR) microscopy. Microplastics of different sizes could be detected and counted in marine sediment samples. The fluorescence staining identified the same particles as those found by scanning a filter area with IR-microscopy.
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                Author and article information

                Contributors
                Role: ConceptualizationRole: Data curationRole: InvestigationRole: SoftwareRole: ValidationRole: Writing – original draft
                Role: ConceptualizationRole: InvestigationRole: SoftwareRole: ValidationRole: Writing – original draft
                Role: InvestigationRole: ValidationRole: Writing – review & editing
                Role: ConceptualizationRole: ResourcesRole: Writing – review & editing
                Role: Funding acquisitionRole: SupervisionRole: Writing – review & editing
                Role: ConceptualizationRole: SupervisionRole: Writing – review & editing
                Role: ConceptualizationRole: Funding acquisitionRole: Project administrationRole: SupervisionRole: Writing – review & editing
                Role: Editor
                Journal
                PLoS One
                PLoS ONE
                plos
                plosone
                PLoS ONE
                Public Library of Science (San Francisco, CA USA )
                1932-6203
                23 June 2020
                2020
                : 15
                : 6
                : e0234766
                Affiliations
                [001]Institute of Hydrochemistry, Chair of Analytical Chemistry and Water Chemistry, Technical University of Munich, Munich, Germany
                University of Montpellier, FRANCE
                Author notes

                Competing Interests: The authors have declared that no competing interests exist

                Author information
                http://orcid.org/0000-0002-7685-5166
                Article
                PONE-D-19-33659
                10.1371/journal.pone.0234766
                7310837
                32574195
                f2c90d8b-df13-4d88-9d19-40ce857a71a5
                © 2020 von der Esch et al

                This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.

                History
                : 5 December 2019
                : 25 May 2020
                Page count
                Figures: 11, Tables: 2, Pages: 20
                Funding
                Funded by: funder-id http://dx.doi.org/10.13039/501100002745, Bayerische Forschungsstiftung;
                Award ID: AZ-1258-16
                Award Recipient :
                Funded by: funder-id http://dx.doi.org/10.13039/501100002347, Bundesministerium für Bildung und Forschung;
                Award ID: 02WRS1378C
                Award Recipient :
                Funding by the Bayerische Forschungsstiftung ( https://www.forschungsstiftung.de/) for the Project MiPaq “Microparticles in the aquatic environment and in foodstuffs” are biodegradable polymers a conceivable solution to the “microplastic problem”?, AZ-1258-16 is gratefully acknowledged. Funding by the German Federal Ministry of Education and Research https://www.bmbf.de/ for the Project MiWa – Microplastic in the water cycle, 02WRS1378C is gratefully acknowledged. Furthermore, this work was supported by the German Research Foundation (DFG, https://www.dfg.de/) and the Technical University of Munich, within the funding program Open Ac-cess Publishing. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript
                Categories
                Research Article
                Computer and Information Sciences
                Computer Software
                Research and Analysis Methods
                Microscopy
                Electron Microscopy
                Scanning Electron Microscopy
                Research and Analysis Methods
                Imaging Techniques
                Fluorescence Imaging
                Research and Analysis Methods
                Microscopy
                Light Microscopy
                Fluorescence Microscopy
                Research and Analysis Methods
                Imaging Techniques
                Image Analysis
                Engineering and Technology
                Signal Processing
                Image Processing
                Physical Sciences
                Chemistry
                Polymer Chemistry
                Macromolecules
                Polymers
                Polycarbonates
                Physical Sciences
                Materials Science
                Materials
                Polymers
                Polycarbonates
                Physical Sciences
                Chemistry
                Polymer Chemistry
                Polymers
                Polycarbonates
                Engineering and Technology
                Signal Processing
                Signal Filtering
                Custom metadata
                Data are in the TUMmedia repository: Kohles AJ, von der Esch E, Anger PM, Hoppe R, Niessner R, Elsner M, et al. TUM-ParticleTyper: Software and Documentation. ( https://mediatum.ub.tum.de/1547636) TUM 2020. doi: 10.14459/2020mp1547636

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