The development and applications of effective analytical techniques for identification and quantification of microplastics in diverse spheres are increasing in the scientific arena. Nile Red (NR) staining has progressed as a low-cost, simple-to-use approach for analyzing the environmental impact of a wide spectrum of microplastics (e.g., ≥ 3 µm - ≤ 5 mm; polyethylene, polypropylene, and polyvinyl chloride etc.). Given the recent surge of research into this methodology, it is critical to examine the findings and present future directions. Herein, we review accomplishments to date of the current protocols describing the sample preparation, staining and fluorescence conditions, contamination measures, and data analysis based on 56 field observations focusing on microplastic pollution and NR staining technique. Additionally, we discuss the challenges in current analyses towards standardization and recommendations related to it. Finally, we conclude that, despite methodological discrepancies, the NR method has emerged as a viable standalone substitute for visual identification; yet not all that fluoresce with NR are microplastics, which necessitates extensive sample preparation or additional spectroscopy techniques for chemical analysis to validate the results. This article informs the reader about how the NR technique is advancing microplastic research and identifies current needs for future advancements.

在科学领域，用于识别和量化不同领域微塑料的有效分析技术的开发和应用正在增加。尼罗红 (NR) 染色已发展成为一种低成本、易于使用的方法，用于分析各种微塑料（例如，≥ 3 µm - ≤ 5 mm；聚乙烯、聚丙烯和聚氯乙烯等）的环境影响.)。鉴于最近对该方法的研究激增，检查结果并提出未来方向至关重要。在此，我们根据 56 次以微塑料污染和 NR 染色技术为重点的现场观察，回顾了描述样品制备、染色和荧光条件、污染措施和数据分析的当前协议迄今为止取得的成就。此外，我们讨论了当前分析中对标准化的挑战以及与之相关的建议。最后，我们得出结论，尽管存在方法上的差异，但 NR 方法已成为视觉识别的可行独立替代品；然而，并非所有与 NR 一起发出荧光的都是微塑料，这需要大量的样品制备或额外的光谱技术进行化学分析以验证结果。本文向读者介绍了 NR 技术如何推进微塑料研究，并确定了当前对未来发展的需求。然而，并非所有与 NR 一起发出荧光的都是微塑料，这需要大量的样品制备或额外的光谱技术进行化学分析以验证结果。本文向读者介绍了 NR 技术如何推进微塑料研究，并确定了当前对未来发展的需求。然而，并非所有与 NR 一起发出荧光的都是微塑料，这需要大量的样品制备或额外的光谱技术进行化学分析以验证结果。本文向读者介绍了 NR 技术如何推进微塑料研究，并确定了当前对未来发展的需求。