Ed For FISH And Live Dead Staining. The 3D Images Were

BMC Microbiology 2010, vtr.org.vn 10:98 website 9 MedChemExpress Amiloride hydrochloride dihydrate ofCOMSTAT analysis of biofilms5.
The thickness function is the maximum thickness over a given location which does not take into account any pores or tour malaysia giá rẻ voids within the biofilm. The thickness distribution is then used to calculate the biofilm roughness and tour malaysia giá rẻ mean biofilm thickness. Roughness coefficient provides an indication of how the thickness of the biofilm varies and Tour singapore malaysia giá rẻ từ hà nội also provides an indication of biofilm heterogeneity [48].Additional file 1: CLSM top view cropped image of S.

oneidensis biofilm (Figure 2) (63? providing a close-up of the nonviable cells using Live/Dead (Baclight) stain. Additional File 1 is a more detailed confocal image of the S. oneidensis biofilm. Its purpose is to show the difference between live and dead cells after using the Live/Dead stain. Additional file 2: Observations of Pure culture continuous time course biofilm study.
A table describing the development of the pure culture biofilms during the continuous experiment. Additional file 3: Observations of Co-culture continuous time course biofilm study. A table describing the development of the coculture biofilms during the continuous experiment.7.8.9.10.11.12.13.14.15.

Acknowledgements This study was supported by the Australian Research Council (grants DP0879245) and The University of Queensland Early Career Researcher Scheme (UQ2006001877). SR is also supported by the Queensland Government (Smart State Award funding), The University of Queensland (Confirmation Scholarship).
P. aeruginosa PAO1, S. oneidensis MR-1 and E.Ed for FISH and Live/Dead staining. The 3D images were created from 1 m z-stacks slices of varying heights (depending on the height of the biofilm) and were constructed using Zeiss 3D imaging software.SEM analysisDuring the continuous experiments one anodic graphite block from each reactor was regularly collected for FISH analysis.

When blocks were initially taken from the reactors, they were washed with basic media that did not include electron donor or acceptor to remove any particulates that may auto fluoresce. FISH sample fixation, hybridization and washing was performed as described previously [41].
Blocks were visualized using the CLSMDuring co-culture experiments blocks (2 mm wide) were removed from the reactors at 72 and 144 hour time points and fixed immediately for SEM analysis. SEM fixation involves the use of 3 solutions. Solution 1 contains 0.043 g lysine (L-lysine free base Sigma L-5501) dissolved in 2 ml of 0.1 M cacodylate buffer.

Solution 2 contains 0.4 ml 25 glutaraldehyde, 1.0 ml 0.2 M cacodylate buffer and 0.6 ml distilled water. Solutions 1 and 2 were mixed together thoroughly immediately before use. Samples were left in this for 10 minutes then transferred to solution 3 which is 2.5 glutaraldehyde in 0.1 M cacodylate buffer for further sample processing as described in Jacques Graham [47].
Samples for SEM were visualized using JEOL JSM- 6400F microscope (10 kV, 3000 V) and EIKO IB-5 sputter coater using platinum.Read et al. BMC Microbiology 2010, 10:98 website 9 ofCOMSTAT analysis of biofilms5. 6.Z-stacks generated using the CLSM were further analysed using COMSTAT to determine roughness coefficient and mean biofilm thickness.

Through COMSTAT a fixed threshold was applied to the images to provide a 0 or 1 value to image pixels.