Integrated SEM, TEM and CLSM workflow for tegumental analysis of adult Schistosoma mansoni.

The tegument of Schistosoma mansoni is a key host-parasite interface and a major target for drug and vaccine development. Although ultrastructural studies of the tegument date back decades, methodological inconsistencies often complicate comparisons across studies. Here, we present a standardized, easy-to-use framework for investigating tegumental architecture using three complementary techniques: scanning electron microscopy (SEM), transmission electron microscopy (TEM), and confocal laser scanning microscopy (CLSM). We provide step-by-step protocols, highlight strengths and limitations of each method, and illustrate their outputs using adult male worms. We also outline a scheme for unbiased quantitative analysis of the resulting images and indicate how the protocols can be adapted to female worms, to other developmental stages and schistosome species, and to immunolocalization at the ultrastructural level. This integrative pipeline enables reproducible morphological characterization and facilitates downstream applications in pharmacology, immunology, and parasite biology.