Influence of the Novel Rapid Preparation Method on Sample Imaging Evaluation for Pathogenic Microorganisms Scanning Electron Microscopy

This study provides a novel rapid and effective sample processing method for scanning electron microscopy observation of bacterium and fungi. We investigated the relationships between image quality and instrument operating parameters. The procedure consists of seven stages, including fixing, cleaning, dehydration, replacement, drying, spraying and observation. The incorporation of tertiary butyl alcohol (TBA), which played important roles in both dehydration and cleaning, was optimized using six distinct tertiary butyl alcohol concentrations (30%, 50%, 70%, 85%, 95% and 100%, v/v). Electrically conductive adhesive transfer tape was chosen as the carrier on which samples were mounted for SEM observation. Saved the time of preparation and cut down the use of expensive toxic reagents. The methodology established in this study helps to improve the efficiency for identifying the ultra microstructures of pathogenic microorganisms. The method describes the complete method development and validation process and is well suitable for pathogenic microorganisms scanning electron microscopy sample imaging evaluation.