title: In Situ Confocal Raman Microscopy of Hydrated Early Stages of Bacterial Biofilm Formation on Various Surfaces in a Flow Cell.
abstract: Bacterial biofilms are precursors to biofouling by other microorganisms. Understanding their initiation may allow us to design better ways to inhibit them, and thus to inhibit subsequent biofouling. In this study, the ability of confocal Raman microscopy to follow the initiation of biofouling by a marine bacterium, Pseudoalteromonas sp. NCIMB 2021 (NCIMB 2021), in a flow cell, using optical and confocal Raman microscopy, was investigated. The base of the flow cell comprised a cover glass. The cell was inoculated and the bacteria attached to, and grew on, the cover glass. Bright field images and Raman spectra were collected directly from the hydrated biofilms over several days. Although macroscopically the laser had no effect on the biofilm, within the first 24 h cells migrated away from the position of the laser beam. In the absence of flow, a buildup of extracellular substances occurred at the base of the biofilm. When different coatings were applied to cover glasses before they were assembled into the flow cells, the growth rate, structure, and composition of the resulting biofilm was affected. In particular, the ratio of Resonance Raman peaks from cytochrome c (CC) in the extracellular polymeric substances, to the Raman phenylalanine (Phe) peak from protein in the bacteria, depended on both the nature of the surface and the age of the biofilm. The ratios were highest for 24 h colonies on a hydrophobic surface. Absorption of a surfactant with an ethyleneoxy chain into the hydrophobic coating created a surface similar to that given with a simple PEG coating, where bacteria grew in colonies away from the surface rather than along the surface, and CC:Phe ratios were initially low but increased at least fivefold in the first 48 h.
authors.affiliation: Department of Chemistry, St. Francis Xavier University, Antigonish, Nova Scotia, Canada tsmithpa@stfx.ca.
authors.forename: Truis
authors.initials: T
authors.lastname: Smith-Palmer
authors.affiliation: Department of Chemistry, St. Francis Xavier University, Antigonish, Nova Scotia, Canada.
authors.forename: Sicheng
authors.initials: S
authors.lastname: Lin
authors.affiliation: Department of Chemistry, St. Francis Xavier University, Antigonish, Nova Scotia, Canada.
authors.forename: Ikenna
authors.initials: I
authors.lastname: Oguejiofor
authors.affiliation: Department of Chemistry, St. Francis Xavier University, Antigonish, Nova Scotia, Canada.
authors.forename: Tianyang
authors.initials: T
authors.lastname: Leng
authors.affiliation: Department of Chemistry, St. Francis Xavier University, Antigonish, Nova Scotia, Canada.
authors.forename: Amanda
authors.initials: A
authors.lastname: Pustam
authors.affiliation: Department of Chemistry, St. Francis Xavier University, Antigonish, Nova Scotia, Canada.
authors.forename: Jin
authors.initials: J
authors.lastname: Yang
authors.affiliation: Department of Biology, St. Francis Xavier University, Antigonish, Nova Scotia, Canada.
authors.forename: Lori L
authors.initials: LL
authors.lastname: Graham
authors.affiliation: Department of Biology, St. Francis Xavier University, Antigonish, Nova Scotia, Canada.
authors.forename: Russell C
authors.initials: RC
authors.lastname: Wyeth
authors.affiliation: Department of Biology, St. Francis Xavier University, Antigonish, Nova Scotia, Canada.
authors.forename: Cory D
authors.initials: CD
authors.lastname: Bishop
authors.affiliation: Department of Biology, St. Francis Xavier University, Antigonish, Nova Scotia, Canada.
authors.forename: M Edwin
authors.initials: ME
authors.lastname: DeMont
authors.affiliation: Department of Physics, St. Francis Xavier University, Antigonish, Nova Scotia, Canada.
authors.forename: David
authors.initials: D
authors.lastname: Pink
mesh_terms: D018441:Biofilms / Q000254:growth & development*; D002465:Cell Movement; D005898:Glass; D018613:Microscopy, Confocal / Q000379:methods*; D011092:Polyethylene Glycols; D031161:Pseudoalteromonas / Q000502:physiology; D013059:Spectrum Analysis, Raman / Q000379:methods*
chemical_list: D011092:Polyethylene Glycols
keywords: Biofilms; Flow cell; Marine; NCIMB 2021; PEG; Polyehtylene glycol; Pseudoalteromonas sp; Raman confocal microscopy