Many bacterial species readily develop biofilms that act as a protective matrix against external challenge, e.g., from antimicrobial treatment. Therefore, biofilms are often responsible for persistent and recurring infections. Established methods for studying biofilms are either destructive or focus on the biofilm’s surface. A non-destructive method that is sensitive to the underside of the biofilm is highly desirable, as it allows studying the penetration of antibiotics through the film. Here, we demonstrate that the high surface sensitivity of resonant hyperspectral imaging provides this capability. The method allows us to monitor the early stages of Escherichia coli biofilm formation, cell attachment and microcolony formation, in-situ and in real-time. We study the response of the biofilm to a number of different antibiotics and verify our observations using confocal microscopy. Based on this ability to closely monitor the surface-bound cells, resonant hyperspectral imaging gives new insights into the antimicrobial resistance of biofilms.

许多细菌物种很容易形成生物膜，以作为抵抗外部挑战（例如，抗微生物处理）的保护性基质。因此，生物膜通常是造成持续和反复感染的原因。研究生物膜的既定方法可能是破坏性的，或者着眼于生物膜的表面。非常需要一种对生物膜底面敏感的非破坏性方法，因为它可以研究抗生素穿过膜的渗透性。在这里，我们证明了共振高光谱成像的高表面灵敏度可提供此功能。该方法使我们可以监测大肠杆菌的早期阶段生物膜形成，细胞附着和微菌落形成，就地和实时。我们研究了生物膜对多种不同抗生素的反应，并使用共聚焦显微镜验证了我们的观察结果。基于这种密切监视表面结合细胞的能力，共振高光谱成像为生物膜的抗微生物性提供了新的见解。