Home›Microbiology›High-resolution Patterned Biofilm Deposition Using pDawn-Ag43
MicrobiologyJoVE (Open Access)Citable · DOI
High-resolution Patterned Biofilm Deposition Using pDawn-Ag43
DOI: 10.3791/58625-v
What you'll learn
✓Prepare pDawn-Ag43 E. coli strains and optical stimulation setup for biofilm patterning
✓Culture spatially patterned bacterial biofilms using light-directed surface adhesion
✓Image and verify high-resolution biofilm deposition patterns
✓Apply optogenetic control to bacterial biofilm engineering and synthetic biology
Protocol
We demonstrate a method for depositing Escherichia coli bacterial biofilms in arbitrary spatial patterns with a high resolution using optical stimulation of a genetically encoded surface-adhesion construct.
Difficulty
advanced
Total time
~3–5 days (strain preparation 1 day, biofilm culture and imaging 2–4 days)
Culture engineered E. coli expressing pDawn-Ag43 adhesin construct and configure projector-based optical illumination system for bacterial pattern stimulation.
▶ 00:57
2
Culture patterned biofilms under light control
Grow bacteria on substrate under programmed spatial light patterns to induce optogenetically controlled biofilm deposition in defined geometries.
▶ 03:44
3
Image deposited biofilm patterns
Acquire microscopy images of patterned biofilms to visualize spatial resolution and fidelity of light-directed deposition.