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Evaluating bacterial spore preparation methods for scanning electron microscopy
Umeå University, Faculty of Science and Technology, Department of Physics.ORCID iD: 0000-0002-0496-6692
Umeå University.ORCID iD: 0000-0002-5773-2793
Umeå University, Faculty of Science and Technology, Department of Physics. Umeå University, Faculty of Medicine, Umeå Centre for Microbial Research (UCMR).ORCID iD: 0000-0002-9835-3263
2024 (English)In: Microscopy and Microanalysis, ISSN 1431-9276, E-ISSN 1435-8115, Vol. 30, no 3, p. 564-573Article in journal (Refereed) Published
Abstract [en]

Scanning electron microscopy (SEM) can reveal the ultrastructure of bacterial spores, including morphology, surface features, texture, spore damage, germination, and appendages. Understanding these features can provide a basis for adherence, how physical and environmental stressors affect spore viability, integrity, and functionality, as well as the distribution and function of surface appendages. However, the spore sample preparation method can significantly impact the SEM images' appearance, resolution, and overall quality. In this study, we compare different spore preparation methods to identify optimal approaches for preparation time, spore appearance and resolved features, including the exosporium and spore pili, for SEM imaging. We use Bacillus paranthracis as model species and evaluate the efficacy of preparation protocols using different fixation and drying methods, as well as imaging under room- and cryogenic temperatures. We compare and assess method complexity to the visibility of the spore exosporium and spore appendages across different methods. Additionally, we use Haralick texture features to quantify the differences in spore surface appearance and determine the most suitable method for preserving spore structures and surface features during SEM evaluation. The findings from this study will help establish protocols for preparing bacterial spores for SEM and facilitating accurate and reliable analysis of spores' characteristics.

Place, publisher, year, edition, pages
Oxford University Press, 2024. Vol. 30, no 3, p. 564-573
Keywords [en]
bacillus, fixative, Haralick, SEM, texture
National Category
Microbiology Other Physics Topics
Identifiers
URN: urn:nbn:se:umu:diva-227941DOI: 10.1093/mam/ozae037ISI: 001217162300001Scopus ID: 2-s2.0-85198180107OAI: oai:DiVA.org:umu-227941DiVA, id: diva2:1884944
Funder
Swedish Research Council, 2019-04016Available from: 2024-07-19 Created: 2024-07-19 Last updated: 2024-07-19Bibliographically approved

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Malyshev, DmitryLee, Cheng ChooAndersson, Magnus

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