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Fluorophore-based Genetically Encoded Biosensors for Ratiometric Fluorescence Imaging in Microbes
Erdem Sunal1, Vanessa Castro-Rodriguez2, Mayuri Sadoine3
1Department of Biology, Heinrich-Heine Universität Düsseldorf.
Journal of Visualized Experiments : Jove
|August 18, 2025
Summary
This study introduces a protocol for using genetically encoded biosensors to track small-molecule dynamics in microbes. This method enhances understanding of microbial function, symbiosis, and disease through quantitative imaging.
Area of Science:
- Microbiology
- Molecular Biology
- Biochemistry
Background:
- Small-molecule dynamics are crucial for microbial physiology, symbiosis, and disease.
- Existing techniques for monitoring these dynamics in vivo are limited.
- Genetically encoded biosensors offer a powerful approach for tracking small molecules.
Purpose of the Study:
- To present a protocol for deploying engineered biosensors into microbes.
- To enable quantitative ratiometric fluorescence imaging of small-molecule dynamics.
- To facilitate broader adoption of biosensor technology in microbial research.
Main Methods:
- Development and implementation of a protocol for genetically encoded biosensor deployment.
- Utilizing quantitative ratiometric fluorescence imaging.
- Employing perfusion devices for controlled environmental conditions.
Main Results:
- A detailed protocol for effective biosensor implementation in microbes.
- Demonstration of high-resolution, spatiotemporal data acquisition.
- Enabling insights into single-cell microbial responses.
Conclusions:
- The presented protocol simplifies the implementation of biosensors for microbial research.
- This approach allows for detailed investigation of small-molecule dynamics.
- It holds significant potential for advancing studies in microbial physiology, symbiosis, and disease.
