As one kind of emerging contaminants, high abundance of microplastics have been identified in ecosystems due to their small size, increasing plastic production, extensive usage, poor waste management, and minimal biological degradation. Therefore, microplastic pollution in the marine and freshwater environment has become a global hot issue in recent years. The most important problem while conducting research on microplastics is to distinguish pollutants from complex environmental media. At present, there are abundant strategies for detection and analysis of microplastics, however, in order to make results comparable, it is urgent to seek unified and standardized separation, extraction, identification and quantification methods for microplastics. Based on a systematic review of common and reliable separation and extraction procedures including density separation, screening, filtration and digestion, elaborating qualitative identification methods such as thermal analysis, spectral analysis and presenting statistical quantification methods such as visual inspection of statistical, fluorescence analysis and thermal analysis, the research prospects of microplastics detection and analysis methods are put forward. Staining fluorescence combined with spectral analysis has broad application prospects in quickly and accurately assessing microplastic contamination in the environment.