File:Oscillations-by-Minimal-Bacterial-Suicide-Circuits-Reveal-Hidden-Facets-of-Host-Circuit-Physiology-pone.0011909.s003.ogv

English: Solid phase growth monitoring of BW25113 cells carrying the ePop plasmid. Microscope slides were prepared by allowing warm LB agarose (1%) containing the appropriate antibiotics and inducers (1mM IPTG, 100ng/uL aTc) to be drawn under a suspended cover slip. After cooling, the coverslip was removed and the agarose pad cut to size with a clean razor blade. 1 µL low density starter culture (BW25113, ePop) was placed on the agarose and a fresh coverslip was applied. Slides were sealed with mineral oil and nailpolish to prevent evaporation. Cells were placed on a Leica inverted microscope in an environmentally controlled enclosure set to 37°C. After 1–2 hours, a region for monitoring was selected on the basis of exhibiting cell division. Images were captured using a Hammamatsu 1384 ORCA-ERA camera at 4 frames per minute overnight using autofocus feature to maintain the focal plane.