A Modified Slide-Embedded Scanning Electron Microscopy Preparation Method to Visualize Antagonistic Interactions Between Trichoderma viride and Fusarium sp.
Year: 2026
Type: article
Abstract: . Fungal interactions occurring directly on the glass surface are subsequently subjected to fixation with 2.5% glutaraldehyde, graded ethanol dehydration, sputter coating, and SEM observation. Compared with conventional methods, the present approach provides improved handling stability, better preservation of native hyphal architecture, reduced deformation during processing, and easier localization of interaction zones during microscopy. The protocol also enables clear visualization of early antagonistic events such as hyphal coiling, penetration, and surface colonization. Due to its simplicity, reproducibility, and minimal technical complexity, this method serves as a practical and efficient approach for SEM-based investigation of fungal-fungal interactions and can be readily adapted for studying diverse mycoparasitic systems. Key features • A simple, cost-effective method to visualize filamentous fungus-fungus interactions, particularly mycoparasitism, using minimal reagents and basic laboratory equipment. • A standardized inoculation strategy with pathogen inoculation 48 h before the mycoparasite ensures reproducible fungal interaction and mycoparasitic development. • Blunt-cut square glass slide pieces embedded in PDA enable easy sample handling, storage, transport, and direct preparation for scanning electron microscopy. • SEM-compatible fixation and ethanol dehydration preserve fungal ultrastructure, enabling high-resolution visualization of hyphal coiling, distortion, and other mycoparasitic interactions.
Source: BIO-PROTOCOL
Authors: Sandra Tomichen, Shweta Panchal
Institutions: Vellore Institute of Technology University, RV University
Language: English
Subfield: Plant Science
Domain: Life Sciences
Open Access status: diamond
Funder: VIT University
OpenAlex ID: W7203444949