Controlling spatial structure in minimal microbial communities by sequential capillary assembly

Area of Science:

• Microbiology
• Bioengineering
• Materials Science

Background:

• Microbial communities exhibit complex social interactions influenced by spatial structure.
• Existing experimental platforms lack the microscale precision needed to control bacterial spatial arrangement and study community development.

Purpose of the Study:

• To develop a high-throughput, micron-scale precision workflow for patterning and growing two bacterial species.
• To enable detailed investigation into the role of initial spatial structure on microbial interactions at low cell densities.

Main Methods:

• Directional sequential capillary assembly of colloidal particles.
• Nanobody-functionalized particles for specific, bio-orthogonal binding reactions between bacteria and surface particles.
• Patterning of *Staphylococcus aureus* and *Escherichia coli* in defined spatial configurations.

Main Results:

• Achieved high-throughput patterning (∼10⁵ cells per template) with micron-scale precision.
• Demonstrated the ability to grow patterned bacterial communities and monitor their development microscopically.
• Established a method for precise spatial control over bacterial interactions.

Conclusions:

• The developed workflow provides a powerful tool for investigating the impact of spatial structure on microbial community dynamics.
• This technique is crucial for understanding and manipulating microbial community development, especially at low cell densities.