Exploiting advances in imaging in microbiology

The funding enabled us to combine the expertise and experience of the two supervisors and groups optimally. This has been invaluable to identify both sample and instrumental limitations and artifacts, and to resolve problems, and to develop robust protocols to image (lab based) bacterial biofilms. The process has enhanced the expertise of both groups. The project has also inspired the named technician to pursue a career in bioimaging. Furthermore, we acquired of a range of custom made hardware enabling us to perform time-lapse live MP-LSCM imaging of biofilms in a flow cell with the full range of objectives. We identified optimal use and settings of the microscope, software and accessories and identified necessary compromises between biological and technical constraints, such as using a high enough laser power to allow visualization of all cells, whilst keeping the biological damage to the minimum. We applied these to obtain high-resolution images of biofilms, and bleach and ablate regions of interest. With this knowledge and newly acquired expertise we are now poised to address questions of significant interest in the biofilm research field, including but not limited to: can a biofilm regenerate after localized internal damage, is resistance to antimicrobials related to microstructure and are cells motile within a biofilm?