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<p>The establishment of microbial biofilms, communities embedded in self-produced extracellular matrices, poses growing challenges for health and antimicrobial management. Understanding biofilm formation is crucial for developing control and eradication strategies. In response to environmental cues, planktonic bacteria adopt a sessile lifestyle, coordinating growth with matrix production. We monitored cellulose biofilm formation by <i>Pseudomonas sp</i>. IsoF in real time using single-step fluorescent stains. Live-tracking of polysaccharide synthesis revealed dynamic matrix arrangements shaping final biofilm structure. Cellulose determined substratum adherence, cell contacts, and colony patterning in IsoF. Biofilms formed in flow-cells and at air-liquid interfaces were remarkably similar in composition, progression, and architecture. Artificial elevation of intracellular c-di-GMP levels produced cellulose-dependent biofilms distinct from the wild type and induced a secondary exopolysaccharide. Our fluorescent probes provide real-time visualization of matrix development, enabling detailed analysis of biofilm architecture and regulation in standard laboratory conditions.</p>

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High-resolution visualization of biofilm matrix development in space and time using fluorescent stains for cellulose

  • 期刊论文

  • 出版模式：

    开放获取

  • 发表日期：

    2026年1月9日

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  • Zaira Heredia-Ponce,
  • Aurélien Bailly,
  • Leo Eberl

摘要

The establishment of microbial biofilms, communities embedded in self-produced extracellular matrices, poses growing challenges for health and antimicrobial management. Understanding biofilm formation is crucial for developing control and eradication strategies. In response to environmental cues, planktonic bacteria adopt a sessile lifestyle, coordinating growth with matrix production. We monitored cellulose biofilm formation by Pseudomonas sp. IsoF in real time using single-step fluorescent stains. Live-tracking of polysaccharide synthesis revealed dynamic matrix arrangements shaping final biofilm structure. Cellulose determined substratum adherence, cell contacts, and colony patterning in IsoF. Biofilms formed in flow-cells and at air-liquid interfaces were remarkably similar in composition, progression, and architecture. Artificial elevation of intracellular c-di-GMP levels produced cellulose-dependent biofilms distinct from the wild type and induced a secondary exopolysaccharide. Our fluorescent probes provide real-time visualization of matrix development, enabling detailed analysis of biofilm architecture and regulation in standard laboratory conditions.