24] Spatial organization of oral bacteria in biofilms

Abstract

This chapter describes the formation of a monospecies oral streptococcal biofilm that is established with only saliva as both the conditioning film and the growth medium. The chapter discusses confocal laser scanning microscopic images obtained of streptococcal biofilms formed in flowcells. The flowcell described in this chapter is especially useful for studying bacterial biofilms because it is easy to manipulate, is camera ready, and is disposable. It allows real time examination of biofilms by placing the flowcell directly on the microscope stage without interrupting the flow dynamics. Because the flow rate is slow, maintenance of the temperature in the flowcell while on the microscope stage is problematic but can be accomplished by passing the fluid through a heating/cooling device just prior to entering the flow cell. Flowcell is used to study the characteristics of colonization of several streptococci. The chapter investigates the accretion of various other oral bacteria onto the initial colonizing streptococci. The combination of fluorescent probes and the flowcell provides an excellent approach for investigating the spatial organization of oral bacterial biofilms.