Rhodamine B dye staining for visualizing microplastics in laboratory-based studies
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RESEARCH ARTICLE
Rhodamine B dye staining for visualizing microplastics in laboratory-based studies Huiyan Tong 1
&
Qianyi Jiang 1 & Xiaocong Zhong 1 & Xingshuai Hu 1
Received: 28 May 2020 / Accepted: 9 September 2020 # Springer-Verlag GmbH Germany, part of Springer Nature 2020
Abstract In the recent years, microplastics have attracted much attention as new emerging environmental pollutants. Previously, several studies were performed to understand the source and fate of microplastics in the environment, organisms, and food webs. To track microplastics and improve their legibility, labeling them is a very effective method during laboratory experiments. This study presents an effective Rhodamine B dye (RhB) staining method for microplastics. The method is crucial for the visual observation of white or transparent plastics by dyeing them in purple or pink, as well as makes the microplastics to fluoresce under common microscope fluorescence filter ranges. Five types of microplastic polymers, namely polyethylene, polypropylene, polystyrene, polyvinyl chloride, and polyurethane were used as the test materials. The efficiencies of ethanol, acetone, and distilled water as possible solvents for dissolving RhB were investigated. Next, the fluorescence stability in various conditions was assessed. The results indicated that ethanol was the most appropriate solvent in dissolving RhB used in staining the microplastics. RhB was fluorescently stable under varying conditions (light and gut fluid) or different solutions (KOH, nitric acid, and saturated NaCl). Additionally, RhB staining exhibited an insignificant effect on the Raman spectra of the microplastics. Our proposed method is simple and robust and helps to visualize the different types of microplastic polymers tested in laboratory experiments, particularly the transparent, white, and small size microplastics. Keywords Rhodamine B dye . Microplastics . Laboratory-based . Staining . Fluorescence
Introduction Plastics are widely used in different sectors, including in the industry, agriculture, and daily life purposes. This is primarily due to their high manufacturing efficiency, enhanced material durability and safety, and low cost of raw materials (Huppertsberg and Knepper 2018). To date, plastics have been used for hundreds of years, and the emergence of plastic products has brought great convenience to human life. However, because of poor management and many other reasons, plastic wastes get their way into the environment, and the external Responsible editor: Philippe Garrigues * Huiyan Tong [email protected] 1
School of Ocean Science and Technology, Dalian University of Technology, No. 2 Dagong Road, New District of Liaodong Bay, Panjin City 124221, Liaoning Province, China
factors like wind, rivers, and ocean currents move the wastes over long distances, of which the plastic waste usually decomposes gradually into either smaller plastic fragments or particles after exposure to long-term physical and chemical effects. At present, microplastics are plastic par
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