Nile Red Staining as a Subsidiary Method for Microplastic Quantifica-tion: A Comparison of Three Solvents and Factors Influencing Application Reliability

Tamminga, Matthias

doi:10.15436/jeses.2.2.1

Cited by 65 publications

(58 citation statements)

References 17 publications

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“…32,48 Other biogenic materials, such as chitin, wood lignin, and natural waxes can stain strongly and may lead to false positive counts when such compounds are abundant. 32,49 Considering that 42-47% of the fibers examined under mFT-IR were cellulose (Table III) and would likely not stain (or stain weakly), a decrease in fiber counts is consistent with our understanding of and experience with Nile red methodology and efficacy. Outdoor fragments likely comprise a variety of plant matter fragments, some of which will stain, so more data are needed to investigate the observed trend in terms of selectivity.…”

Section: Nile Red Quantificationsupporting

confidence: 67%

“…Beyond the ability to selectively stain plastics, we have found Nile red improves counts of smaller-sized fragments by creating a clearer fluorescing signal on a dark background that is more easily visible and counted, something observed by other researchers as well. 32,34,49 Maes et al 32 predict that fragments down to 5 mm may potentially be visualized, depending upon the optics of the microscope in use. Further, Nile red will illuminate clear and white fragments that are obscured by the GF (or any other light colored) filter background.…”

Section: Nile Red Quantificationmentioning

confidence: 99%

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Microplastics Differ Between Indoor and Outdoor Air Masses: Insights from Multiple Microscopy Methodologies

et al. 2020

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The abundance and distribution of microplastic (<5 mm) has become a growing concern, particularly over the past decade. Research to date has focused on water, soil, and organism matrices but generally disregarded air. We explored airborne microplastic inside and outside of buildings in coastal California by filtering known volumes of air through glass fiber filters, which were then subsequently characterized with a variety of microscopy techniques: gross traditional microscopy, fluorescent microscopy following staining with Nile red, micro-Raman spectroscopy, and micro-Fourier transform infrared (µFT-IR) spectroscopy. Microplastics permeated the air, with indoor (3.3 ± 2.9 fibers and 12.6 ± 8.0 fragments m–3; mean ± 1 SD) harboring twice as much as outdoor air (0.6 ± 0.6 fibers and 5.6 ± 3.2 fragments m–3). Microplastic fiber length did not differ significantly between indoor and outdoor air, but indoor microplastic fragments (58.6 ± 55 µm) were half the size of outdoor fragments (104.8 ± 64.9 µm). Micro-Raman and FT-IR painted slightly different pictures of airborne plastic compounds, with micro-Raman suggesting polyvinyl chloride dominates indoor air, followed by polyethylene (PE) and µFT-IR showing polystyrene dominates followed by PE and polyethylene terephthalate. The ubiquity of airborne microplastic points to significant new potential sources of plastic inputs to terrestrial and marine ecosystems and raises significant concerns about inhalation exposure to humans both indoors and outdoors.

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Section: Nile Red Quantificationsupporting

confidence: 67%

Section: Nile Red Quantificationmentioning

confidence: 99%

Microplastics Differ Between Indoor and Outdoor Air Masses: Insights from Multiple Microscopy Methodologies

et al. 2020

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“…has been suggested in the literature; e.g., acetone, chloroform, n-hexane (Tamminga, 2017;Wiggin and Holland, 2019) toluene, cyclohexane, ethanol, ethylacetate, acetonitrile, dichloromethane (Shim et al, 2016); methanol (Shim et al, 2016;Erni-Cassola et al, 2017). Furthermore, solvents such as dimethyl sulfoxide (DMSO), glycerol, and ethylene diaminotetraacetic acid (EDTA) have been used to dissolve NR while staining neutral lipids (Satpati and Pal, 2015;Alemán-Nava et al, 2016).…”

Section: Discussionmentioning

confidence: 99%

Preliminary Results From Detection of Microplastics in Liquid Samples Using Flow Cytometry

et al. 2020

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Microplastics are globally recognized as contaminants in freshwater and marine aquatic systems. To date there is no universally accepted protocol for isolation and quantification of microplastics from aqueous media. Various methodologies exist, many of which are time consuming and have the potential to introduce contaminants into samples, thereby obscuring characterization of the environmental microplastic load. Here, we present first steps in the detection of microplastics in liquid samples, based on their fluorescent staining followed by high throughput analysis and quantification using Flow Cytometry. Using controlled laboratory settings nine polymer types [polystyrene (PS); polyethylene (PE); polyethylene terephthalate (PET/PETE); high density polyethylene (HDPE); low density polyethylene (LDPE); polyvinyl chloride (PVC); polypropylene (PP); nylon (PA); polycarbonate (PC)] were tested for identification and quantification in freshwater. All nine plastic types were stained with 10 µg/mL Nile Red in 10% dimethyl sulfoxide with a 10 min incubation time. The lowest spatial detectable limit for plastic particles was 200 nm. Out of the nine polymer types chosen for the study PS, PE, PET, and PC were well-identified; however, results for other plastic types (PVC, PP, PA, LDPE, and HDPE) were masked to certain extent by Nile Red aggregation and precipitation. The methodology presented here permits identification of a range of particle sizes and types. It represents a significant step in the quantification of microplastics by replacing visual data interpretation with a sensitive and automated method.

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“…A frequently applied technique is staining with Nile Red. This choice for a specific dye has been based on testing a range of alternatives, combined with several solvents [17,82,83]. Nile Red is solvatochromic, resulting in a red-shift of its fluorescence emission spectrum related to an increasing polarity of the material surrounding the Nile Red molecules.…”

Section: Detection Techniquesmentioning

confidence: 99%

Current Insights into Monitoring, Bioaccumulation, and Potential Health Effects of Microplastics Present in the Food Chain

Raamsdonk

Zande

Koelmans

et al. 2020

Foods

169

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Microplastics (MPs) are considered an emerging issue as environmental pollutants and a potential health threat. This review will focus on recently published data on concentrations in food, possible effects, and monitoring methods. Some data are available on concentrations in seafood (fish, bivalves, and shrimps), water, sugar, salt, and honey, but are lacking for other foods. Bottled water is a considerable source with numbers varying between 2600 and 6300 MPs per liter. Particle size distributions have revealed an abundance of particles smaller than 25 µm, which are considered to have the highest probability to pass the intestinal border and to enter the systemic circulation of mammals. Some studies with mice and zebrafish with short- or medium-term exposure (up to 42 days) have revealed diverse results with respect to both the type and extent of effects. Most notable modifications have been observed in gut microbiota, lipid metabolism, and oxidative stress. The principal elements of MP monitoring in food are sample preparation, detection, and identification. Identified data gaps include a lack of occurrence data in plant- and animal-derived food, a need for more data on possible effects of different types of microplastics, a lack of in silico models, a lack of harmonized monitoring methods, and a further development of quality assurance.

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Nile Red Staining as a Subsidiary Method for Microplastic Quantifica-tion: A Comparison of Three Solvents and Factors Influencing Application Reliability

Cited by 65 publications

References 17 publications

Microplastics Differ Between Indoor and Outdoor Air Masses: Insights from Multiple Microscopy Methodologies

Microplastics Differ Between Indoor and Outdoor Air Masses: Insights from Multiple Microscopy Methodologies

Preliminary Results From Detection of Microplastics in Liquid Samples Using Flow Cytometry

Current Insights into Monitoring, Bioaccumulation, and Potential Health Effects of Microplastics Present in the Food Chain

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