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Three-dimensional Confocal Raman Imaging of Secreted Chemical Signals in Pseudomonas Aeruginosa Biofilms
Thursday, March 11, 2021: 8:30 AM - 8:50 AM
Speaker(s)
Description
The use of confocal Raman imaging is becoming increasingly popular with its versality to perform both two-dimensional (2D) scan and z-depth profiling on microbiological samples to gather abundant molecular and structural information. Pseudomonas aeruginosa is an opportunistic human pathogen related to acute and chronic diseases, and the complex biofilm matrix it forms has evidenced to be closely related to the concerning antibiotic resistance that makes the treatment difficult. Within P. aeruginosa quorum sensing (QS) signal system, a family of signaling molecules, alkyl quinolones (AQs), is secreted and used to coordinate various physiological behaviors including biofilm formation. In this study, we built up a three-dimensional (3D) confocal Raman imaging platform to analyze the molecular and spatial distribution of AQ molecules in P. aeruginosa biofilms. We started with a model polymer system to characterize the robustness of the technique. As we applied the technique to bacteria biofilms, we found that early-stage biofilms (8 h) displayed the of 2-Heptyl-4-hydroxyquinoline N-oxide (HQNO) and 2-Heptyl-3-hydroxy-4-quinolone (PQS), and they were detected in different locations and depths. Depth profiling images also showed that AQs can be consistently detected vertically across 3-5 μm within biofilms. Furthermore, mature biofilms (24 h) tends to form larger aggregations that can stretch over 10 μm. Bright-field optical images of the biofilms are used to confirmed the continuous growth of biofilms at different stages at a macroscopic level. From the 2D and 3D Raman images of P. aeruginosa biofilms, we are able to visualize and analyze the spatiotemporal distributions of different AQs that is critical to the QS-regulated biofilm formation, and demonstrated how these factors come in play regarding the complex biofilm formation of P. aeruginosa communities that will be useful for tacking the treatment of biofilm-related infections and diseases.
Additional Info
Keywords: Please select up to 4 keywords ONLY:
Data Analysis and Manipulation,Microscopy,Raman/Surface Enhanced Raman Spectroscopy,Spectroscopy
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