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DropSOAC: Stabilizing Microfluidic Drops for Time-Lapse Quantification of Single-Cell Bacterial Physiology
Shawna L Pratt1,2, Geoffrey K Zath1,2, Tatsuya Akiyama1,3
1Center for Biofilm Engineering, Montana State University, Bozeman, MT, United States.
Frontiers in Microbiology
|October 15, 2019
Summary
We developed DropSOAC, a microfluidic method to stabilize single bacterial cells in drops for over 20 hours. This allows high-throughput study of microbial physiology and growth heterogeneity in populations.
Area of Science:
- Microbiology
- Biotechnology
- Biophysics
Background:
- Microbial populations exhibit physiological heterogeneity, contributing to resilience against stressors like antibiotics.
- Single-cell analysis is crucial for understanding subpopulations responsible for stress survival and community regeneration.
- Existing microfluidic methods face challenges with drop stability during long-term imaging.
Purpose of the Study:
- To develop and validate a novel microfluidic technique for stabilizing single-cell bacterial cultures in drops for extended periods.
- To enable high-throughput, time-lapse imaging and analysis of bacterial growth heterogeneity at the single-cell level.
- To characterize physiological differences in bacterial subpopulations, such as growth rates and lag times.
Main Methods:
- Utilized drop-based microfluidics to encapsulate individual bacterial cells in growth medium droplets.
- Developed a device preparation strategy (DropSOAC) involving soaking in a water-oil reservoir to achieve thermodynamic equilibrium.
- Employed confocal laser scanning microscopy (CLSM) for time-lapse imaging of bacterial growth within stabilized microfluidic drops.
- Studied *Pseudomonas aeruginosa* and its Δ*hpf* mutant during resuscitation after starvation.
Main Results:
- Achieved stable drop position and volume for over 20 hours using the DropSOAC method.
- Successfully characterized growth rate and lag time heterogeneity of hundreds of individual bacterial cells.
- Demonstrated the ability to analyze physiological differences between single cells within a population starting from isolated cells.
Conclusions:
- The DropSOAC method significantly enhances the stability of microfluidic drops for long-term single-cell imaging.
- This approach provides a high-throughput platform for investigating microbial physiology and heterogeneity at the single-cell level.
- DropSOAC facilitates the characterization of physiological variations within bacterial populations, crucial for understanding resilience and adaptation.
