AIMS: To investigate the use of confocal Raman microspectroscopy (CRM) for the analysis of the structure, composition and development of fully hydrated biofilms. METHODS AND RESULTS: Pseudomonas aeruginosa PAO1 biofilms were cultured in a flow cell in minimal nutrient medium (artificial sea water) and their development was followed for up to 3 weeks. The spectroscopic signature of the biofilm cells and extracellular polymeric substances (EPS) were differentiated and their distribution in biofilm colonies and within water channels was mapped in-plane and -depth. The colonies were initially amorphous, mainly composed of cells with no detectable amount of EPS. They developed rapidly to give round colonies composed of a cellular core enclosed in a sheath of EPS. The EPS continued to increase and spread throughout the biofilm to become the dominating feature of aged colonies. Colonies with a liquid core morphology - characteristic of the seeding dispersal process - were also observed. CONCLUSIONS: This study demonstrated that CRM can be used to monitor the distribution of biofilm components in fully hydrated undisturbed biofilms over time. SIGNIFICANCE AND IMPACT OF THE STUDY: Confocal Raman microspectroscopy facilitates the analysis of hydrated, live bacterial biofilms as a function of space and time, thus making it a suitable technique for investigating the effects of various additives and environmental factors on biofilm growth.
AIMS: To investigate the use of confocal Raman microspectroscopy (CRM) for the analysis of the structure, composition and development of fully hydrated biofilms. METHODS AND RESULTS:Pseudomonas aeruginosa PAO1 biofilms were cultured in a flow cell in minimal nutrient medium (artificial sea water) and their development was followed for up to 3 weeks. The spectroscopic signature of the biofilm cells and extracellular polymeric substances (EPS) were differentiated and their distribution in biofilm colonies and within water channels was mapped in-plane and -depth. The colonies were initially amorphous, mainly composed of cells with no detectable amount of EPS. They developed rapidly to give round colonies composed of a cellular core enclosed in a sheath of EPS. The EPS continued to increase and spread throughout the biofilm to become the dominating feature of aged colonies. Colonies with a liquid core morphology - characteristic of the seeding dispersal process - were also observed. CONCLUSIONS: This study demonstrated that CRM can be used to monitor the distribution of biofilm components in fully hydrated undisturbed biofilms over time. SIGNIFICANCE AND IMPACT OF THE STUDY: Confocal Raman microspectroscopy facilitates the analysis of hydrated, live bacterial biofilms as a function of space and time, thus making it a suitable technique for investigating the effects of various additives and environmental factors on biofilm growth.
Authors: Oscar D Ayala; Catherine A Wakeman; Isaac J Pence; Jennifer A Gaddy; James C Slaughter; Eric P Skaar; Anita Mahadevan-Jansen Journal: ACS Infect Dis Date: 2018-06-25 Impact factor: 5.084
Authors: Nameera Baig; Sneha Polisetti; Nydia Morales-Soto; Sage J B Dunham; Jonathan V Sweedler; Joshua D Shrout; Paul W Bohn Journal: Proc SPIE Int Soc Opt Eng Date: 2016-09-27
Authors: Gaddi Blumrosen; Daniela Vecchio; Saiqa I Khan; Alexander Golberg; Michael C McCormack; Martin L Yarmush; Michael R Hamblin; William G Austen Journal: Biotechnol Bioeng Date: 2015-09-09 Impact factor: 4.530
Authors: Ota Samek; Katarina Mlynariková; Silvie Bernatová; Jan Ježek; Vladislav Krzyžánek; Martin Šiler; Pavel Zemánek; Filip Růžička; Veronika Holá; Martina Mahelová Journal: Int J Mol Sci Date: 2014-12-22 Impact factor: 5.923