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.
The bacterial biofilm is a microbial community in which bacteria are embedded in the extracellular matrix and can also be used as a solid composite. It was found that internal stresses are generated during pellicle growth, which exists between the air and the liquid. But we do not know if there is the internal stress in the biofilm, which exists between the air and the solid, and how does the internal stress evolve and distribute in the growing biofilm. So, in this paper, we make the growing substrate into the micropillar array to grow biofilms, each micropillar has the deformation due to the growing heterogeneity of the biofilm around the micropillar, and we can get the internal stress by measuring each micropillar’s deformation. First, we find that the direction of the internal stress is approximately along the biofilm expansion at the early time, colonies are formed in the biofilm at the later time, which cause the internal stress locally along the expansion of the colony. Second, the internal stress is proportional to the biofilm thickness. Finally, we find that the matrix producing cells contribute more the internal stress, and the internal stress evolving is closely related to the secretion of the extracellular matrix. Form our work, we obtain the distribution of the internal stress direction, we also can use the biofilm thickness, which is easy to measure, express the internal stress approximately, by doing so, we can further study other phenomena of biofilms, such as self-healing and mechanical resistance.
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