Université Paris-Saclay, INRAE, AgroParisTech, Micalis Institute, Jouy-en-Josas, France.
Sorbonne Université, CNRS, IBPS Institute, Paris, France.
Methods Mol Biol. 2025;2852:159-170. doi: 10.1007/978-1-0716-4100-2_11.
The functional properties of biofilms are intimately related to their spatial architecture. Structural data are therefore of prime importance to dissect the complex social and survival strategies of biofilms and ultimately to improve their control. Confocal laser scanning microscopy (CLSM) is the most widespread microscopic tool to decipher biofilm structure, enabling noninvasive three-dimensional investigation of their dynamics down to the single-cell scale. The emergence of fully automated high content screening (HCS) systems, associated with large-scale image analysis, has radically amplified the flow of available biofilm structural data. In this contribution, we present a HCS-CLSM protocol used to analyze biofilm four-dimensional structural dynamics at high throughput. Meta-analysis of the quantitative variables extracted from HCS-CLSM will contribute to a better biological understanding of biofilm traits.
生物膜的功能特性与其空间结构密切相关。因此，结构数据对于剖析生物膜复杂的社会和生存策略至关重要，最终有助于提高对生物膜的控制。共聚焦激光扫描显微镜 (CLSM) 是解析生物膜结构最广泛使用的显微镜工具，能够对其动态进行非侵入式的三维研究，深入到单细胞尺度。全自动高通量筛选 (HCS) 系统的出现，结合大规模图像分析，极大地增加了可用生物膜结构数据的数量。在本研究中，我们提出了一种 HCS-CLSM 方案，用于高通量分析生物膜的四维结构动力学。对 HCS-CLSM 提取的定量变量进行的荟萃分析将有助于更好地理解生物膜特性的生物学基础。
