Three-Dimensional Imaging and Quantification of Biomass and Biofilms in Porous Media

A new method to resolve biofilms in three dimensions in porous media using high-resolution synchrotron-based x-ray computed microtomography (CMT) has been developed. Imaging biofilms in porous media without disturbing the natural spatial arrangement of the porous media and associated biofilm has been a challenging task, primarily because porous media generally precludes conventional imaging via optical microscopy; x-ray tomography offers a potential alternative. One challenge for using this method is that most conventional x-ray contrast agents are water-soluble and easily diffuse into biofilms. To overcome this problem, silver-coated microspheres were added to the fluid phase to create an x-ray contrast that does not diffuse into the biofilm mass. Using this approach, biofilm imaging in porous media was accomplished with sufficient contrast to differentiate between the biomass- and fluid-filled pore spaces. The method was validated by using a two-dimensional micro-model flow cell where both light microscopy and CMT imaging were used to im age the biofilm. The results of this work has been published in Water Resources Research (Iltis et al., 2010). Additional work needs to be done to optimize this imaging approach, specifically, we find that the quality of the images are highly dependent on the coverage of the biofilm with Ag particles, - which means that we may have issues in dead-end pore space and for very low density (fluffy) biofilms. What we can image for certain with this technique is the biofilm surface that is well-connected to flow paths and thus well-supplied with nutrients etc