Abstract
ABSTRACTCapturing microbial growth on a macroscopic scale is of great importance to further our understanding of microbial life. However, methods for imaging microbial life on a scale of millimeters to centimeters are often limited by designs that have poor environmental control, resulting in dehydration of the agar plate within just a few days. Here, we created MOCHA (microbial chamber), a simple but effective chamber that allows users to study microbial growth for extended periods (weeks) in a stable environment. Agar hydration is maintained with a double-decker design, in which two glass petri dishes are connected by a wick, allowing the lower plate to keep the upper plate hydrated. This flexible chamber allows the observation of a variety of microbiological phenomena, such as the growth and development of single bacterial and fungal colonies, interspecies interactions, swarming motility, and pellicle formation.IMPORTANCE Detailed study of microbial life on the colony scale of millimeters to centimeters has been lagging considerably behind microscopic inspection of microbes. One major reason for this is the lack of inexpensive instrumentation that can reproducibly capture images in a controlled environment. In this study, we present the design and use of a unique chamber that was used to produce several time-lapse movies that aimed to capture the diversity of microbial colony phenotypes over long periods.
Highlights
Capturing microbial growth on a macroscopic scale is of great importance to further our understanding of microbial life
Microbes display a staggering variety of growth phenotypes on the macroscopic scale, including sectoring, swarming, biofilm formation, pellicle formation, and changes in morphology due to chemical or environmental signaling
In order to demonstrate the improvements gained with the use of MOCHA, two separate time-lapse movies were obtained
Summary
Capturing microbial growth on a macroscopic scale is of great importance to further our understanding of microbial life. Great progress has been achieved in the microscopic inspection of single-celled life, increasing our capacity to visualize cells to 20-nm resolution using superresolution microscopy [3] Despite this progress, capturing with time-lapse photography the diversity and dynamic biology of microbial interactions at the macroscale of millimeters to centimeters has lagged considerably. One example is the large microbial evolution and growth arena (MEGA) plate, which is used to monitor emerging bacterial mutants in response to increasing levels of antibiotics on a large agar plate [5] This method captures microbial growth over time, it is not temperature controlled, it can support colonies for only 12 days, and it has limited resolution for discerning details of colony morphologies, due to the large size of the agar plate (120 cm long). We hope that the dissemination of such a design will facilitate the evolution of improved apparatuses for capturing novel microbial phenotypes on the macroscopic scale
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