Instituto de Hortofruticultura Subtropical y Mediterránea "La Mayora", Universidad de Málaga-Consejo Superior de Investigaciones Científicas (IHSM-UMA-CSIC), Málaga, Spain.
Departamento de Microbiología, Universidad de Málaga, Málaga, Spain.
Microbiol Spectr. 2022 Aug 31;10(4):e0093922. doi: 10.1128/spectrum.00939-22. Epub 2022 Jul 5.
Complex interactions between microbial populations can greatly affect the overall properties of a microbial community, sometimes leading to cooperation and mutually beneficial coexistence, or competition and the death or displacement of organisms or subpopulations. Interactions between different biofilm populations are highly relevant in diverse scientific areas, from antimicrobial resistance to microbial ecology. The utilization of modern microscopic techniques has provided a new and interesting insight into how bacteria interact at the cellular level to form and maintain microbial biofilms. However, our ability to follow complex intraspecies and interspecies interactions at the microscopic level has remained somewhat limited. Here, we detailed BacLive, a novel noninvasive method for tracking bacterial growth and biofilm dynamics using high-resolution fluorescence microscopy and an associated ImageJ processing macro (https://github.com/BacLive) for easier data handling and image analysis. Finally, we provided examples of how BacLive can be used in the analysis of complex bacterial communities. Communication and interactions between single cells are continuously defining the structure and composition of microbial communities temporally and spatially. Methods routinely used to study these communities at the cellular level rely on sample manipulation which makes microscopic time-lapse experiments impossible. BacLive was conceived as a method for the noninvasive study of the formation and development of bacterial communities, such as biofilms, and the formation dynamics of specialized subpopulations in time-lapse experiments at a colony level. In addition, we developed a tool to simplify the processing and analysis of the data generated by this method.
微生物种群之间的复杂相互作用会极大地影响微生物群落的整体特性，有时会导致合作和互利共生，或者竞争和生物体或亚群的死亡或取代。不同生物膜种群之间的相互作用在从抗菌耐药性到微生物生态学的多个科学领域都具有高度相关性。现代微观技术的利用为我们提供了一个新的有趣视角，了解细菌如何在细胞水平上相互作用以形成和维持微生物生物膜。然而，我们在微观水平上跟踪复杂种内和种间相互作用的能力仍然有些有限。在这里，我们详细介绍了 BacLive，这是一种使用高分辨率荧光显微镜跟踪细菌生长和生物膜动态的新型非侵入性方法，以及一个相关的 ImageJ 处理宏（https://github.com/BacLive），用于更轻松地处理数据和图像分析。最后，我们提供了一些示例，说明如何在分析复杂细菌群落时使用 BacLive。单细胞之间的交流和相互作用不断地定义微生物群落的结构和组成。在细胞水平上研究这些群落的常规方法依赖于样本处理，这使得微观时程实验成为不可能。BacLive 的构思是一种非侵入性方法，用于研究细菌群落（如生物膜）的形成和发展，以及在菌落水平的时程实验中专门的亚群形成动态。此外，我们开发了一种工具，用于简化该方法生成的数据的处理和分析。
