Dependence of the Bacillus subtilis biofilm expansion rate on phenotypes and the morphology under different growing conditions

Biofilms are communities of tightly associated bacteria encased in an extracellular matrix and attached to surfaces of various objects, such as liquid or solid surfaces. Here we use the multi-channel wide field stereo fluorescence microscope to characterize growth of the Bacillus subtilis biofilm, in which the bacterial strain was triple fluorescence labeled for three main phenotypes: motile, matrix producing and sporulating cells. We used the feature point matching approach analyzing time lapse experimental movies to study the biofilm expansion rate. We found that the matrix producing cells dominate the biofilm expansion, at the biofilm edge, the expansion rate of matrix producing cells was almost the same as the velocity of the whole biofilm; however, the motile and sporulating cells were nearly rest. We also found that the biofilm expansion rate evolution relates to cell differentiation and biofilm morphology, and other micro-environments can influence the biofilm growth, such as nutrient, substrate hardness and colony competition. From our work, we get a deeper understanding of the biofilm growth, which can help us to control and to further disperse the biofilm.