Methods for biofilm constituents and turnover

Abstract : Sensitive, rapid and specific methods for monitoring development and turnover are extremely important for a comprehensive understanding of biofilms. The first part of this chapter describes methods of Multimodal Laser Scanning Microscopy (ML-SM) for in situ identification of biofilm components and monitoring of biofilm development, pattern of gene expression, and visualization of dynamic molecular processes in biofilms. The second part gives an overview of designing and characterizing luminescent reporter systems for high-throughput screening of bioactive molecules. This part specifically addresses the GacS/GacA two-component regulatory system, which is central to biofilm formation in all γ-proteobacteria, and any well understood regulatory cascade can be targeted with a similar approach by targeting important promoters in the pathway. Methods described in this chapter are useful in screening compounds and their libraries for antibacterial, quorum sensing and biofilm inhibitory compounds.