The morphotype M of S. marcescens is a derivative of F. It was obtained after many repeated attempts to grow the F morphotype in suspensions in the minimal medium MM. E. coli strain 281 was obtained from the collection of the Department of Genetics and Microbiology, Faculty of Sciences, Charles University. Cultivation If not specified otherwise, bacteria were grown at NAG at 27°C in sealed boxes with controlled humidity. Stabilates were kept at −80°C [20]. New colonies were initiated as follows: (1) as clones from single cells, by classical sowing of bacterial suspension (in phosphate buffer); (2)

planted by dropping dense suspension (108/ml) on a defined place (diameter about 2 mm); (3) planted by dotting from material taken by a sterile needle from an older Nutlin-3a research buy body; (4) by smearing (to grow maculae): 30 μl of bacterial suspension (approx. 108 cells) was applied to a line of approx. 5 cm. For conditioned agar see [3]. Documentation Plates were photographed in situ using Olympus

C-5050ZOOM digital camera under VX-680 price ambient or penetrating light (Fomei, LP-400 light panel, cold cathode light) or under magnification using a binocular magnifier [3]. Colony margins were observed with fully motorized microscope stand IX81 (Olympus) equipped with objectives LUCPLFLN 20 (NA 0.45) and LUCPLFLN 40 (NA 0.60) and documented with the camera HAMMATSU Orca, with differential interference contrast. Digital images were further elaborated by the software Olympus CELL^R SYSTEM. STK38 Figures shown were selected from an extensive collection of primary photos from several repetitions Selleck ATM Kinase Inhibitor (5 and more) of each experiment. Photoshop software was used to assemble the plates as they

appear in Figures. No image doctoring was performed except automatic adjustment of brightness and contrast in some cases. Acknowledgements Supported by the Grant Agency of Czech Republic 408/08/0796 (JČ, IP, AB, AM, ZN), and by the Czech Ministry of education MSM 0021620845 (AM, AB, ZN). The authors thank Josef Lhotsky for invaluable comments, Alexander Nemec for strain determination, and Ondřej Šebesta for assistance with microscopy. References 1. Aguilar C, Vlamakis H, Losick R, Kolter R: Thinking about Bacillus subtilis as a multicellular organism. Curr Opin Microbiol 2007, 10:638–43.PubMedCrossRef 2. Ben-Jacob E, Levine H: Self-engineering capabilities of bacteria. J R Soc Interface 2005, 3:197–214.CrossRef 3. Čepl JJ, Pátková I, Blahůšková A, Cvrčková F, Markos A: Patterning of mutually interacting bacterial bodies: close contacts and airborne signals. BMC Microbiol 2010, 10:139.PubMedCrossRef 4. Shapiro JA: Bacteria are small but not stupid: cognition, natural genetic engineering and socio-bacteriology. Stud Hist Phil Biol Biomed Sci 2007, 38:807–819. 5. Shapiro JA: Bacteria as multicellular organism. In Multicellularity: The rule, not the exception. Lessons fromE.colicolonies. Edited by: Dworkin M, Shapiro JA. University Press, Oxford; 1997:14–49. 6.