An Applied Framework of CLSM and Rheology for Controlling Foodborne Biofilms and Their Cleaning Agents and Disinfectants Resistance: A Review

Biofilms formed by foodborne pathogens pose a significant threat to food safety, as they enhance microbial resistance to disinfectants and environmental stress. Understanding the relationship between their intricate structure and macroscopic mechanical properties is crucial for developing effective control strategies. This review systematically investigates how the combined application of confocal laser scanning microscopy (CLSM) and rheology provides a powerful solution to this problem. CLSM technology reveals biofilm structure and extracellular polymeric substance (EPS) distribution through high-resolution 3D imaging and multi-component fluorescence labeling. Rheology quantitatively analyzes key mechanical parameters such as viscoelasticity and yield stress. Their integration establishes a multidimensional "structure-function-mechanics" research framework, enabling cross-scale correlations from microscopic features to macroscopic mechanical responses. Additionally, this review analyzes current biofilm removal strategies, fluorescent labeling methods for EPS components, and the composition and functions of EPS in typical bacterial biofilms. We further explore optimized technical approaches to overcome food matrix interference, applications in cleaning process optimization, and future standardization directions integrated with artificial intelligence. This comprehensive methodology provides a systematic and efficient paradigm for understanding biofilm mechanisms and developing control strategies, holding significant implications for biofilm prevention in the food industry.