Modification of a Nile Red Staining Method for Microplastics Analysis: A Nile Red Plate Method

Recently, environmental pollution from microplastics (MPs) has become a significant reason for increasing the number of studies to develop analysis methods. The Nile red staining method (NR-S), which is staining polymer particles with Nile red (NR) dye, has been widely used for the analysis of MPs in environmental samples. However, this method has several limitations, as it is difficult to stain MPs covered with organic matter residues. In this study, we modified the NR-S method into an NR plate method (NR-P), where the plate is coated with NR instead of staining MPs directly. The optimum concentration of NR solution was obtained (1000 mg/L), and the effectiveness of the NR-P method for the analysis of MPs was assessed using different types (polypropylene, polyethylene, polyethylene terephthalate, and polystyrene), sizes (100–1000 µm), and shapes (sphere, fiber, film, and flake) of plastic materials. The NR-P method demonstrated improved resolution in the overall types, shapes, and sizes of MPs and was better than the control (without NR plate method) and NR-S method. In particular, the NR-P method can effectively observe MPs covered with organic matter, which was a major limitation of the NR-S method. Finally, MPs in sewage field samples were analyzed by the NR-P method with an accuracy of 78% confirmed by FT-IR. We demonstrated that this method is a convenient and efficient alternative for identifying MPs, even for field samples.