Soft X-ray microscopy (SXM) using synchrotron radiation is a newly-available technology for high resolution imaging of hydrated specimens, making it potentially valuable for the study of biofilms. In this research, SXM was used to investigate bacterial spatial distribution and biofilm structure during the early stages of colonization of an exposed surface. Pseudomonas putida DMP-1 formed a closely-packed monolayer on a silicon nitride substratum after 24 hours of growth. The initial development of a multilayer, interlocking structure was observed. The soft X-ray images were taken with the XM-1 transmission X-ray microscope, located at the Center for X-ray Optics, Lawrence Berkeley National Laboratory. The results indicate the potential of SXM to provide new insights for the study of biofilms.

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