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.
KeywordsBiofilm Pathogenic microorganisms Sample preparation SEM
We are grateful to Canquan Mao Professor from Laboratory of Molecular Evolution and Applied Biology, School of Life Science and Engineering, Southwest Jiaotong University, Chengdu, P. R. China for helping us with language editing and research platform. This work was financially supported by Sichuan Provincial Department of Education 2017 Natural Science Key Project (No. 17ZA0095); Sichuan Provincial Key Laboratory of Medicinal and Edible Plant Resources Development Fund Project (No. 10Y201705).
Conflict of Interest.
Mingzhu Song and Xirui Wang contributed equally to this work, as co-author of the first author. All authors have no conflicts of interest.
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