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Selection of microplastics by Nile Red staining increases environmental sample throughput by micro-Raman spectroscopy
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This study showed that pre-selecting particles with Nile Red staining prior to Raman spectroscopy analysis significantly increases sample throughput, enabling faster processing of environmental microplastic samples without sacrificing identification accuracy.
Nile Red staining enables visual identification and quantification of fluorescent particles as a proxy to microplastics at low cost and high throughput, including those of small sizes (≥2 μm), when preceded by proper natural organic matter removal, but providing no chemical characterization. On the other hand, micro-spectroscopy methods allow chemical characterization of particles based on their spectra, essential for polymer identification, but are costly and time-consuming. This work addresses the combination of both Nile Red staining with micro-Raman spectroscopy for the identification of microplastics. Besides being useful for quantification, Nile Red staining can be advantageously used as an objective criterion for pre-selection of particles for micro-Raman spectroscopy, producing little interference. The use of the 442 nm laser in micro-Raman spectroscopy induces Nile Red luminescence thus allowing to target the specific suspected microplastics when using an orange filter, reducing the number of particles subjected to identification and improving sample throughput. Staining dyes could also be used for mapping suspected microplastics before targeted analysis by micro-Raman spectroscopy. Thus, coupling Nile Red with micro-Raman spectroscopy can be useful to improve time efficiency while using this equipment.
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Nile red staining in microplastic analysis—proposal for a reliable and fast identification approach for large microplastics
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Researchers tested Nile red staining with UV light photography as a rapid and reliable method for identifying large microplastics in environmental samples, finding it comparable to fluorescence microscopy-based staining and confirmed by μ-Raman spectroscopy.
Optimization of sample preparation, fluorescence- and Raman techniques for environmental microplastics
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Researchers optimized methods for preparing and analyzing environmental microplastic samples using fluorescence staining with Nile Red dye and Raman spectroscopy. The study found that while fluorescence can broadly categorize plastics as polar or non-polar, Raman spectroscopy with a deep-UV laser was needed to reliably identify all polymer types, including those pigmented with carbon black.
Identification and Quantification of Microplastics Using Nile Red Staining
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This study developed a rapid fluorescent staining method using the dye Nile Red to identify and quantify microplastics in environmental samples, offering an alternative to slower and more expensive spectroscopic techniques. Nile Red staining provides a faster way to differentiate microplastics from natural particles in large sample sets, improving throughput for environmental monitoring.
The assessment of particle selection and blank correction to enhance the analysis of microplastics with Raman microspectroscopy
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Researchers improved the efficiency of Raman microspectroscopy analysis for microplastics by implementing automated particle selection and optimized blank correction methods, significantly reducing analysis time while maintaining accuracy for complex environmental samples.
Fast microplastics identification with stimulated Raman scattering microscopy
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Stimulated Raman scattering microscopy was applied to rapidly identify and image microplastic particles in complex environmental samples at speeds dramatically faster than conventional Raman spectroscopy. The technique has potential to enable high-throughput microplastic analysis that could make large-scale environmental monitoring more feasible.
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