Journal article
Kinetic Visualization of Single-Cell Interspecies Bacterial Interactions
JOVE-JOURNAL OF VISUALIZED EXPERIMENTS, Vol.2020(162), pp.1-18
08/01/2020
DOI: 10.3791/61376
PMID: 32831301
Abstract
Polymicrobial communities are ubiquitous in nature, yet studying their interactions at the single-cell level is difficult. Thus, a microscopy-based method has been developed for observing interspecies interactions between two bacterial pathogens. The use of this method to interrogate interactions between a motile Gram-negative pathogen, Pseudomonas aeruginosa and a non-motile Gram-positive pathogen, Staphylococcus aureus is demonstrated here. This protocol consists of co-inoculating each species between a coverslip and an agarose pad, which maintains the cells in a single plane and allows for visualization of bacterial behaviors in both space and time.
Furthermore, the time-lapse microscopy demonstrated here is ideal for visualizing the early interactions that take place between two or more bacterial species, including changes in bacterial species motility in monoculture and in coculture with other species. Due to the nature of the limited sample space in the microscopy setup, this protocol is less applicable for studying later interactions between bacterial species once cell populations are too high. However, there are several different applications of the protocol which include the use of staining for imaging live and dead bacterial cells, quantification of gene or protein expression through fluorescent reporters, and tracking bacterial cell movement in both single species and multispecies experiments.
Details
- Title: Subtitle
- Kinetic Visualization of Single-Cell Interspecies Bacterial Interactions
- Creators
- Kaitlin D. Yarrington - Roy J. and Lucille A. Carver College of MedicineAndrea Sánchez Peña - Roy J. and Lucille A. Carver College of MedicineDominique H. Limoli - Roy J. and Lucille A. Carver College of Medicine
- Resource Type
- Journal article
- Publication Details
- JOVE-JOURNAL OF VISUALIZED EXPERIMENTS, Vol.2020(162), pp.1-18
- Publisher
- Journal Of Visualized Experiments
- DOI
- 10.3791/61376
- PMID
- 32831301
- ISSN
- 1940-087X
- eISSN
- 1940-087X
- Number of pages
- 18
- Grant note
- 5T32HL007638-34 / NIH T32 Training Grant; United States Department of Health & Human Services; National Institutes of Health (NIH) - USA LIMOLI19R3 / Cystic Fibrosis Foundation Junior Faculty Recruitment Award LIMOLI18F5 / Cystic Fibrosis Foundation Postdoc-to-Faculty Transition Award
- Language
- English
- Date published
- 08/01/2020
- Academic Unit
- Microbiology and Immunology
- Record Identifier
- 9984297327702771
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