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Real-time Imaging and Quantification of Fungal Biofilm Development Using a Two-Phase Recirculating Flow System
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Linking biofilm spatial structure to real-time microscopic oxygen decay imaging
S Rubol1, A Freixa2, X Sanchez-Vila3
1a Department of Energy Resources Engineering , Stanford University , Stanford , CA , USA.
Biofouling
|February 7, 2018
Summary
Researchers combined confocal laser scanning microscopy (CLSM) and planar optode imaging to link aquatic biofilm structure to oxygen levels. This integration reveals how biofilm architecture influences oxygen decay at the microscale.
Area of Science:
- Aquatic microbiology
- Biofilm research
- Microbial ecology
Background:
- Biofilms are complex microbial communities.
- Understanding biofilm structure-function relationships is crucial.
- Oxygen dynamics are key metabolic indicators in biofilms.
Purpose of the Study:
- To integrate confocal laser scanning microscopy (CLSM) and planar optode imaging.
- To correlate fine-scale biofilm structure with real-time oxygen distribution.
- To investigate the impact of light and temperature on biofilm architecture and metabolism.
Main Methods:
- Non-destructive imaging techniques: CLSM and VisiSens planar optodes.
- Biofilm cultivation under contrasting light and temperature conditions (10/20°C) for seven days.
- Geo-statistical analysis of CLSM images to quantify aggregate sizes and distribution.
Main Results:
- Biofilm structures comprised small (~10^0 μm) and medium (~10^1 μm) irregular aggregates.
- Cyanobacteria and extracellular polymeric substances (EPS) formed larger aggregates in dark-grown biofilms.
- Algal aggregates were larger in light-exposed, 20°C biofilms.
- Light-exposed, 20°C biofilms exhibited higher density and sparser structure with lower respiration rates.
- A positive correlation was observed between occupied pixel count and oxygen decay rate.
Conclusions:
- The combined CLSM and optode approach, utilizing geo-statistics, effectively links biofilm architecture to metabolism at the micrometric scale.
- Environmental factors like light and temperature significantly influence biofilm structural organization and oxygen dynamics.
- This integrated methodology offers a promising tool for detailed microscale biofilm analysis.
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