Interstitial Biofilm Formation in P. Aeruginosa (data for Supplementary 3, Figure 3G in related publication): time series images

This folder contains 1000 tiff images. These tif images comprise a time series captured at a rate of 1 frame every 2 seconds. These images were captured using an inverted microscope with a 100x phase oil-immersion objective (Olympus IX-71) fitted with an FViewII (Soft Imaging Systems) monochromatic camera.

Selected frames from this movie were used in the creation of Figure 3G in the related publication, showing phase-contrast images of the assembly of a small bulldozer aggregate breaking away from an established trail in an interstitial biofilm to form a new trail that intersects with another newly formed trail. (Scale bar, 5 μm.)

Interstitial biofilms of the bacteria Pseudomonas aeruginosa were cultured on 0.4xLB-Lennox solidified with 8 g/L gellan gum (twitching motility gellan gum, TMGG). Molten TMGG was poured over sterile slides and solidified at room temperature. Slides were inoculated, covered with a sterile coverslip, and incubated in humid conditions at 37 °C. Media was supplemented with 100 Kunitz units per mL DNaseI (D5025, Sigma Aldrich) or the enzyme storage buffer [50% vol/vol glycerol, 10 mM MgCl2, 10 mM CaCl2, 10 mM Tris•HCl].