Crayfish is one of the most important freshwater aquaculture species in China. The potential risks of crayfish consumption caused by environmental microplastic pollution have attracted much attention. In this study, a total of 72 crayfish samples were exposed to the microplastic concentrations of 1 mg/L, 3 mg/L, and 9 mg/L for 7, 14, and 28 days, and microplastic contamination levels in crayfish were then explored by laser confocal micro-Raman (LCM-Raman) imaging and scanning electron microscope (SEM). LCM-Raman imaging showed better performance in microplastics identification. Besides, the average percentage of the contaminated area in visualized LCM-Raman images was used to quantitatively assess contamination levels. Following 28 days of exposure to 9 mg/L microplastics, microplastic accumulation reached about 13,000 particles per crayfish. The results confirmed that LCM-Raman imaging combined with image processing technology could be used to construct a high-performance analytical strategy for the assessment of microplastic contamination in crayfish.