Microbial single-cell applications under anoxic conditions

Keating, Ciara; Fiege, Kerstin; Diender, Martijn; Sousa, Diana Z.; Villanueva, Laura

Abstract

The field of microbiology traditionally focuses on studying microorganisms at the population level. Nevertheless, the application of single-cell level methods, including microfluidics and imaging techniques, has revealed heterogeneity within populations, making these methods essential to understand cellular activities and interactions at a higher resolution. Moreover, single-cell sorting has opened new avenues for isolating cells of interest from microbial populations or complex microbial communities. These isolated cells can be further interrogated in downstream single-cell “omics” analyses, providing physiological and functional information. However, applying these methods to study anaerobic microorganisms under in situ conditions remains challenging due to their sensitivity to oxygen. Here, we review the existing methodologies for the analysis of viable anaerobic microorganisms at the single-cell level, including live-imaging, cell sorting, and microfluidics (lab-on-chip) applications, and we address the challenges involved in their anoxic operation. Additionally, we discuss the development of non-destructive imaging techniques tailored for anaerobes, such as oxygen-independent fluorescent probes and alternative approaches.

Citation

Keating, C., Fiege, K., Diender, M., Sousa, D. Z., & Villanueva, L. (2024). Microbial single-cell applications under anoxic conditions. Applied and Environmental Microbiology, 90(11), Article e01321-24. https://doi.org/10.1128/aem.01321-24

• Journal Article Type: Article
• Acceptance Date: Aug 29, 2024
• Online Publication Date: Sep 30, 2024
• Publication Date: 2024-11
• Deposit Date: Oct 16, 2024
• Publicly Available Date: Oct 17, 2024
• Journal: Applied and Environmental Microbiology
• Print ISSN: 0099-2240
• Electronic ISSN: 1098-5336
• Publisher: American Society for Microbiology
• Peer Reviewed: Peer Reviewed
• Volume: 90
• Issue: 11
• Article Number: e01321-24
• DOI: https://doi.org/10.1128/aem.01321-24
• Public URL: https://durham-repository.worktribe.com/output/2960887