CLSM images of dual-species C. albicans SC5314-S. gordonii biofilms formed on saliva-coated glass cover slips in YPT-G medium. Biofilms were grown as described in Methods for 6 h at 37°C and fluorescently stained with FITC. For each of the panels (A–D) a series of x-y sections were collected to generate 3D reconstructions of representative CLSM images with Volocity® software. Heights were measured across ten CLSM image stacks from two independent experiments ± SD. Panels: (a), C. albicans-S.gordonii DL1, height 81.4 ± 7.50 μm; (b), C. albcians-S. gordonii ΔfruRBA, height 113 ± 10.4 μm*; (c), C. albicans-S. gordonii ΔfruRBA/pfruRBA, height 70.6 ± 6.40 μm; (d), C. albicans, height 86.8 ± 11.6. *P<0.001 when heights were compared using the t test. Note that in (b) there is more substratum vacant and more hyphae are visible compared with the other dual-species biofilm panels (a) and (c).