TY - JOUR
T1 - A Versatile Strategy for Characterization and Imaging of Drip Flow Microbial Biofilms
AU - Li, Bin
AU - Dunham, Sage J.B.
AU - Ellis, Joseph F.
AU - Lange, Justin D.
AU - Smith, Justin R.
AU - Yang, Ning
AU - King, Travis L.
AU - Amaya, Kensey R.
AU - Arnett, Clint M.
AU - Sweedler, Jonathan V.
N1 - Funding Information:
The authors would like to acknowledge intellectual contributions from Troy Comi and Elizabeth Neumann. SEM support was provided by Catherine Wallace and Scott Robinson from the Beckman Institute Imaging Technology Group, which is partially supported by the National Science Foundation Award No. DBI-9871103. Funding for this study was provided through a cooperative agreement with the Engineer Research and Development Center−Construction Engineering Research Laboratory under agreement number W9132T-15-2-0006. J.F.E. and S.J.B.D. gratefully acknowledge support from the Springborn Endowment, and S.J.B.D. received partial financial support through the NSF-GRFP. Although this research was sponsored by the Department of Defense, the content of the information does not necessary reflect the position or policy of the government, and no official endorsement should be inferred.
PY - 2018/6/5
Y1 - 2018/6/5
N2 - The inherent architectural and chemical complexities of microbial biofilms mask our understanding of how these communities form, survive, propagate, and influence their surrounding environment. Here we describe a simple and versatile workflow for the cultivation and characterization of model flow-cell-based microbial ecosystems. A customized low-shear drip flow reactor was designed and employed to cultivate single and coculture flow-cell biofilms at the air-liquid interface of several metal surfaces. Pseudomonas putida F1 and Shewanella oneidensis MR-1 were selected as model organisms for this study. The utility and versatility of this platform was demonstrated via the application of several chemical and morphological imaging techniques - including matrix-assisted laser desorption/ionization mass spectrometry imaging, secondary ion mass spectrometry imaging, and scanning electron microscopy - and through the examination of model systems grown on iron substrates of varying compositions. Implementation of these techniques in combination with tandem mass spectrometry and a two-step imaging principal component analysis strategy resulted in the identification and characterization of 23 lipids and 3 oligosaccharides in P. putida F1 biofilms, the discovery of interaction-specific analytes, and the observation of several variations in cell and substrate morphology present during microbially influenced corrosion. The presented workflow is well-suited for examination of both single and multispecies drip flow biofilms and offers a platform for fundamental inquiries into biofilm formation, microbe-microbe interactions, and microbially influenced corrosion.
AB - The inherent architectural and chemical complexities of microbial biofilms mask our understanding of how these communities form, survive, propagate, and influence their surrounding environment. Here we describe a simple and versatile workflow for the cultivation and characterization of model flow-cell-based microbial ecosystems. A customized low-shear drip flow reactor was designed and employed to cultivate single and coculture flow-cell biofilms at the air-liquid interface of several metal surfaces. Pseudomonas putida F1 and Shewanella oneidensis MR-1 were selected as model organisms for this study. The utility and versatility of this platform was demonstrated via the application of several chemical and morphological imaging techniques - including matrix-assisted laser desorption/ionization mass spectrometry imaging, secondary ion mass spectrometry imaging, and scanning electron microscopy - and through the examination of model systems grown on iron substrates of varying compositions. Implementation of these techniques in combination with tandem mass spectrometry and a two-step imaging principal component analysis strategy resulted in the identification and characterization of 23 lipids and 3 oligosaccharides in P. putida F1 biofilms, the discovery of interaction-specific analytes, and the observation of several variations in cell and substrate morphology present during microbially influenced corrosion. The presented workflow is well-suited for examination of both single and multispecies drip flow biofilms and offers a platform for fundamental inquiries into biofilm formation, microbe-microbe interactions, and microbially influenced corrosion.
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U2 - 10.1021/acs.analchem.8b00560
DO - 10.1021/acs.analchem.8b00560
M3 - Article
C2 - 29723465
AN - SCOPUS:85046676705
VL - 90
SP - 6725
EP - 6734
JO - Analytical Chemistry
JF - Analytical Chemistry
SN - 0003-2700
IS - 11
ER -