Figure 1

Growth and development of Haloferax volcanii static liquid biofilms. (A) Cells within typical shaken culture of H. volcanii DS2 under transmitted light. Scale bar equals 10 μm. (B) Cross section of cryo-processed H. volcanii colony biofilm grown on CA medium for 5days and stained with CellMask Orange (CMO). Scale bar equals 30 μm. (C) Photographs of SL-biofilms routinely grown and analyzed within chamber slides (top left; scale bar equals 2 mm) and on glass coupons within six-well plates (bottom left; scale bar equals 1 cm). A macroscopic photograph of biofilm growth on a 12.7-mm glass coupon is shown in the center (scale bar equals 2 mm) with an area magnified on the right (shown in white box). (D-H) CLSM of biofilms grown on glass coupons (within a six-well plate in Hv-Ca medium; scale bars equal 30 μm). Biofilms stained with FM 1-43 were imaged directly in bulk Hv-starve medium using a 63× water-immersion objective after 2 days (D), 5 days (E) and 7 days (F-H). (I) Biofilm development shown through orthogonal views of SL-biofilms on glass coupons stained with CMO. CLSM, confocal laser scanning microscopy; CMO, CellMask Orange; SL-biofilm, static liquid biofilm.