The pervasive use of plastics has transformed modern life, but plastic pollution poses significant environmental challenges, mainly through microplastics (MPs). This chapter discusses advanced techniques for collecting, separating, and identifying microplastics in environmental samples, which is essential for understanding their ecological impacts. With global plastic production projected to reach 600 million tons by 2025, the accumulation of microplastics in freshwater and marine ecosystems is alarming. MPs arise from both primary sources, such as industrial pellets, and secondary sources, resulting from the degradation of larger plastics. Their environmental persistence leads to bioaccumulation in aquatic organisms, causing various physiological and ecological harm. This chapter outlines innovative methods for isolating microplastics from environmental matrices, including density-based separation for sediments, size-based filtration separation, and enzymatic digestion for biological samples. Additionally, microscopic analysis techniques are highlighted for characterizing microplastics, allowing researchers to identify their shape, size, and polymer composition. Advanced characterization techniques, including Fourier-transform infrared spectroscopy (FTIR) and Raman spectroscopy, are presented to identify polymeric composition and particle size. Future perspectives emphasize the need for standardized sampling protocols and the integration of automated monitoring systems to achieve reliable data on microplastic concentrations. Innovations in remote sensing and analytical technologies promise enhanced detection and quantification capabilities. By addressing the complexities of microplastic pollution, this chapter aims to provide a comprehensive framework for researchers and policymakers to understand better and mitigate the environmental impacts of microplastics.