Šmerdová Lenka, Füzik Tibor, Valentová Lucie, Bárdy Pavol, Procházková Michaela, Pařenicová Martina, Plevka Pavel
Central European Institute of Technology, Masaryk University, Brno, Czech Republic.
Veterinary Research Institute, Brno, Czech Republic.
Biochem Biophys Rep. 2025 Aug 16;43:102127. doi: 10.1016/j.bbrep.2025.102127. eCollection 2025 Sep.
Biofilm formation exacerbates bacterial infections and interferes with industrial processes. However, the dynamics of biofilm development, especially if formed by a combination of more than one species, is not entirely understood. Here, we present a microfluidic cultivation system that enables continuous imaging of biofilm growth using light sheet fluorescence microscopy (LSFM). We studied the development of biofilms of the human pathogens and . Multidirectional LSFM imaging enables the calculation of a three-dimensional reconstruction of the biofilm structure with isotropic resolution. Whereas forms 50-70-μm-thick mushroom-like structures, a biofilm is 10-15 μm thick with cell clusters 25 μm in diameter. A combined biofilm resulted in the formation of large mushroom-like clusters of cells that were subsequently dispersed by invading A higher inoculation ratio favoring resulted in the formation of small and stable clusters overgrown with cells. Applying conditioned media from and coculture to a single-species biofilm induced its dispersion. Integrating a microfluidic system into LSFM enables the visualization of biofilm formation dynamics and the effects of compounds on biofilm development.
生物膜的形成会加剧细菌感染并干扰工业生产过程。然而，生物膜形成的动态过程，尤其是由多种细菌共同形成的生物膜的动态过程，尚未完全明晰。在此，我们展示了一种微流控培养系统，该系统能够使用光片荧光显微镜（LSFM）对生物膜生长进行连续成像。我们研究了人类病原体 和 的生物膜形成过程。多方向LSFM成像能够以各向同性分辨率计算生物膜结构的三维重建。 形成50 - 70微米厚的蘑菇状结构，而 生物膜厚度为10 - 15微米，有直径为25微米的细胞簇。混合生物膜导致形成大的 细胞蘑菇状簇，随后这些簇被入侵的 分散。有利于 的更高接种比例导致形成小而稳定的 簇，并被 细胞过度生长。将 和 共培养的条件培养基应用于单物种 生物膜会诱导其分散。将微流控系统与LSFM相结合能够可视化生物膜形成动态以及化合物对生物膜发育的影响。
