期刊
JOVE-JOURNAL OF VISUALIZED EXPERIMENTS
卷 -, 期 152, 页码 -出版社
JOURNAL OF VISUALIZED EXPERIMENTS
DOI: 10.3791/60275
关键词
Immunology and Infection; Issue 152; antibiotics; antimicrobials; co-culture; inhibition assays; microbial interactions; phenotypic screen
资金
- University of Wisconsin-Madison, Office of the Vice Chancellor for Research and Graduate Education
- Wisconsin Alumni Research Foundation
- National Institutes of Health Centers for Excellence for Translational Research [U19-AI109673-01]
- National Library of Medicine training grant [NLM 5T15LM007359]
The study of interactions between microorganisms has led to numerous discoveries, from novel antimicrobials to insights in microbial ecology. Many approaches used for the study of microbial interactions require specialized equipment and are expensive and time intensive. This paper presents a protocol for co-culture interaction assays that are inexpensive, scalable to large sample numbers, and easily adaptable to numerous experimental designs. Microorganisms are cultured together, with each well representing one pairwise combination of microorganisms. A test organism is cultured on one side of each well and first incubated in monoculture. Subsequently, target organisms are simultaneously inoculated onto the opposite side of each well using a 3D-printed inoculation stamp. After co-culture, the completed assays are scored for visual phenotypes, such as growth or inhibition. These assays can be used to confirm phenotypes or identify patterns among isolates of interest. Using this simple and effective method, users can analyze combinations of microorganisms rapidly and efficiently. This co-culture approach is applicable to antibiotic discovery as well as culture-based microbiome research and has already been successfully applied to both applications.
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