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. 2017 Dec 19;7:17864. doi: 10.1038/s41598-017-17721-3

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© The Author(s) 2017

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.

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Films of bacteria at interfaces. (a) Low hexadecane toxicity for PAO1 and PA14 cells. Cells were re-suspended in saline solution (with or without a minimal media supplement, MMS) and exposed to hexadecane for 2 or 40 days and analyzed for viability by counting colony forming units (CFU) on an agar plate after incubation for 24 h. Cells remained viable for 40 days. (b) A PDMS platform with 50 micron diameter pores was fabricated to observe PAO1 and PA14 cells confined at the oil-water interface in the absence of nutrient. The cells display a differential response to the interface. PAO1 cells aggregate to form the “chef hat” structure. PA14 cells form an active layer characterized by a highly motile phase (scale bar: 20 μm). (c) FBI (Film of Bacteria at Interfaces) formation for P. aeruginosa PAO1 cells including the interaction of the bacteria with the oil droplet, adhesion, accumulation of the cells on the oil droplet, and maturation of the film. Single cells adhere to and cover the fluid interface over time. Interfaces are completely covered by a complex structure after 10 days (scale bar: 20 μm, 20 μm, and 1 mm, respectively) as cells adhere and accumulate as they secrete structures that form a film. (d) Scanning electron microscopy images of interfaces aged for 10 days reveal an asymmetric structure of PAO1 cells within a matrix of extracellular material (scale bar: 10 μm and 2 μm for low- and high-magnification images).