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Proceedings Paper

Label-free molecular imaging of bacterial communities of the opportunistic pathogen Pseudomonas aeruginosa
Author(s): Nameera Baig; Sneha Polisetti; Nydia Morales-Soto; Sage J. B. Dunham; Jonathan V. Sweedler; Joshua D. Shrout; Paul W. Bohn
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Paper Abstract

Biofilms, such as those formed by the opportunistic human pathogen Pseudomonas aeruginosa are complex, matrix enclosed, and surface-associated communities of cells. Bacteria that are part of a biofilm community are much more resistant to antibiotics and the host immune response than their free-floating counterparts. P. aeruginosa biofilms are associated with persistent and chronic infections in diseases such as cystic fibrosis and HIV-AIDS. P. aeruginosa synthesizes and secretes signaling molecules such as the Pseudomonas quinolone signal (PQS) which are implicated in quorum sensing (QS), where bacteria regulate gene expression based on population density. Processes such as biofilms formation and virulence are regulated by QS. This manuscript describes the powerful molecular imaging capabilities of confocal Raman microscopy (CRM) and surface enhanced Raman spectroscopy (SERS) in conjunction with multivariate statistical tools such as principal component analysis (PCA) for studying the spatiotemporal distribution of signaling molecules, secondary metabolites and virulence factors in biofilm communities of P. aeruginosa. Our observations reveal that the laboratory strain PAO1C synthesizes and secretes 2-alkyl-4-hydroxyquinoline N-oxides and 2-alkyl-4-hydroxyquinolones in high abundance, while the isogenic acyl homoserine lactone QS-deficient mutant (ΔlasIΔrhlI) strain produces predominantly 2-alkyl-quinolones during biofilm formation. This study underscores the use of CRM, along with traditional biological tools such as genetics, for studying the behavior of microbial communities at the molecular level.

Paper Details

Date Published: 27 September 2016
PDF: 8 pages
Proc. SPIE 9930, Biosensing and Nanomedicine IX, 993004 (27 September 2016); doi: 10.1117/12.2236695
Show Author Affiliations
Nameera Baig, Univ. of Notre Dame (United States)
Sneha Polisetti, Univ. of Notre Dame (United States)
Nydia Morales-Soto, Univ. of Notre Dame (United States)
Sage J. B. Dunham, Univ. of Illinois at Urbana-Champaign (United States)
Jonathan V. Sweedler, Univ. of Illinois at Urbana-Champaign (United States)
Joshua D. Shrout, Univ. of Notre Dame (United States)
Paul W. Bohn, Univ. of Notre Dame (United States)

Published in SPIE Proceedings Vol. 9930:
Biosensing and Nanomedicine IX
Hooman Mohseni; Massoud H. Agahi; Manijeh Razeghi, Editor(s)

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