Label-free molecular imaging of bacterial communities of the opportunistic pathogen Pseudomonas aeruginosa

Baig, Nameera; Polisetti, Sneha; Morales-Soto, Nydia; Dunham, Sage J. B.; Sweedler, Jonathan V.; Shrout, Joshua D.; Bohn, Paul W.

doi:10.1117/12.2236695

Label-free molecular imaging of bacterial communities of the opportunistic pathogen Pseudomonas aeruginosa

Journal Article · Tue Sep 27 04:00:00 EDT 2016 · Proceedings of SPIE - The International Society for Optical Engineering

DOI:https://doi.org/10.1117/12.2236695· OSTI ID:1807684

Baig, Nameera ^[1]; Polisetti, Sneha ^[2]; Morales-Soto, Nydia ^[2]; Dunham, Sage J. B. ^[3]; Sweedler, Jonathan V. ^[3]; Shrout, Joshua D. ^[2]; Bohn, Paul W. ^[2]

Univ. of Notre Dame (United States); University of Notre Dame
Univ. of Notre Dame (United States)
Univ. of Illinois at Urbana-Champaign (United States)

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. Furthermore, 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.

View Accepted Manuscript (DOE)

Research Organization:: Univ. of Notre Dame, IN (United States)

Sponsoring Organization:: USDOE; National Institutes of Health (NIH)

Grant/Contract Number:: AC05-00OR22725

OSTI ID:: 1807684

Journal Information:: Proceedings of SPIE - The International Society for Optical Engineering, Journal Name: Proceedings of SPIE - The International Society for Optical Engineering Vol. 9930; ISSN 0277-786X

Publisher:: SPIECopyright Statement

Country of Publication:: United States

Language:: English

References (26)

Smith, Richard J. Biopolymers https://doi.org/10.1002/(ISSN)1097-0282	journal
Cover Picture Petry, Renate; Schmitt, Michael; Popp, Jurgen ChemPhysChem, Vol. 4, Issue 1 https://doi.org/10.1002/cphc.v4:1	journal	January 2003
Dueling quorum sensing systems in Pseudomonas aeruginosa control the production of the Pseudomonas quinolone signal (PQS) McGrath, Stephen; Wade, Dana S.; Pesci, Everett C. FEMS Microbiology Letters, Vol. 230, Issue 1 https://doi.org/10.1016/S0378-1097(03)00849-8	journal	January 2004
4-Quinolone signalling in Pseudomonas aeruginosa: Old molecules, new perspectives Diggle, Stephen P.; Cornelis, Pierre; Williams, Paul International Journal of Medical Microbiology, Vol. 296, Issue 2-3 https://doi.org/10.1016/j.ijmm.2006.01.038	journal	April 2006
Culture-free diagnostics of Pseudomonas aeruginosa infection by silver nanorod array based SERS from clinical sputum samples Wu, Xiaomeng; Chen, Jing; Li, Xibo Nanomedicine: Nanotechnology, Biology and Medicine, Vol. 10, Issue 8 https://doi.org/10.1016/j.nano.2014.04.010	journal	November 2014
Vibrational spectroscopic characterization of fluoroquinolones Neugebauer, U.; Szeghalmi, A.; Schmitt, M. Spectrochimica Acta Part A: Molecular and Biomolecular Spectroscopy, Vol. 61, Issue 7 https://doi.org/10.1016/j.saa.2004.11.014	journal	May 2005
Vibrational spectra study on quinolones antibiotics Wang, Yu; Yu, Ke; Wang, Sihuan Spectrochimica Acta Part A: Molecular and Biomolecular Spectroscopy, Vol. 65, Issue 1 https://doi.org/10.1016/j.saa.2005.09.043	journal	September 2006
Pyocyanin effects on respiratory epithelium: relevance in Pseudomonas aeruginosa airway infections Rada, Balázs; Leto, Thomas L. Trends in Microbiology, Vol. 21, Issue 2 https://doi.org/10.1016/j.tim.2012.10.004	journal	February 2013
MALDI-guided SIMS: Multiscale Imaging of Metabolites in Bacterial Biofilms Lanni, Eric J.; Masyuko, Rachel N.; Driscoll, Callan M. Analytical Chemistry, Vol. 86, Issue 18 https://doi.org/10.1021/ac5020222	journal	August 2014
Adsorption and surface-enhanced Raman of dyes on silver and gold sols Lee, P. C.; Meisel, D. The Journal of Physical Chemistry, Vol. 86, Issue 17, p. 3391-3395 https://doi.org/10.1021/j100214a025	journal	August 1982
Label-Free in Situ SERS Imaging of Biofilms Ivleva, Natalia P.; Wagner, Michael; Szkola, Agathe The Journal of Physical Chemistry B, Vol. 114, Issue 31 https://doi.org/10.1021/jp102466c	journal	July 2010
Three-Dimensional Hierarchical Plasmonic Nano-Architecture Enhanced Surface-Enhanced Raman Scattering Immunosensor for Cancer Biomarker Detection in Blood Plasma Li, Ming; Cushing, Scott K.; Zhang, Jianming ACS Nano, Vol. 7, Issue 6 https://doi.org/10.1021/nn4018284	journal	May 2013
Correlated mass spectrometry imaging and confocal Raman microscopy for studies of three-dimensional cell culture sections Ahlf, Dorothy R.; Masyuko, Rachel N.; Hummon, Amanda B. The Analyst, Vol. 139, Issue 18 https://doi.org/10.1039/C4AN00826J	journal	January 2014
Multimodal chemical imaging of molecular messengers in emerging Pseudomonas aeruginosa bacterial communities Baig, Nameera F.; Dunham, Sage J. B.; Morales-Soto, Nydia The Analyst, Vol. 140, Issue 19 https://doi.org/10.1039/C5AN01149C	journal	January 2015
Illuminating disease and enlightening biomedicine: Raman spectroscopy as a diagnostic tool Ellis, David I.; Cowcher, David P.; Ashton, Lorna The Analyst, Vol. 138, Issue 14 https://doi.org/10.1039/c3an00698k	journal	January 2013
Correlated imaging – a grand challenge in chemical analysis Masyuko, Rachel; Lanni, Eric J.; Sweedler, Jonathan V. The Analyst, Vol. 138, Issue 7 https://doi.org/10.1039/c3an36416j	journal	January 2013
Biofilm formation at the air-liquid interface by the Pseudomonas fluorescens SBW25 wrinkly spreader requires an acetylated form of cellulose: Biofilm formation by P. fluorescens SBW25 Spiers, Andrew J.; Bohannon, John; Gehrig, Stefanie M. Molecular Microbiology, Vol. 50, Issue 1 https://doi.org/10.1046/j.1365-2958.2003.03670.x	journal	August 2003
Confocal Raman microspectroscopy as a tool for studying the chemical heterogeneities of biofilms in situ: Biofilm Raman microspectroscopy Sandt, C.; Smith-Palmer, T.; Pink, J. Journal of Applied Microbiology, Vol. 103, Issue 5 https://doi.org/10.1111/j.1365-2672.2007.03413.x	journal	June 2007
The Pseudomonas quinolone signal (PQS), and its precursor HHQ, modulate interspecies and interkingdom behaviour: Quinolone signal molecules modulate interkingdom behaviour Reen, F. Jerry; Mooij, Marlies J.; Holcombe, Lucy J. FEMS Microbiology Ecology, Vol. 77, Issue 2 https://doi.org/10.1111/j.1574-6941.2011.01121.x	journal	May 2011
Quinolones: from antibiotics to autoinducers Heeb, Stephan; Fletcher, Matthew P.; Chhabra, Siri Ram FEMS Microbiology Reviews, Vol. 35, Issue 2 https://doi.org/10.1111/j.1574-6976.2010.00247.x	journal	March 2011
MicroCommentary Tian, Miaoying; Day, Brad Molecular Microbiology, Vol. 61, Issue 5 https://doi.org/10.1111/mmi.2006.61.issue-5	journal	September 2006
MicroCommentary Husson, Robert N. Molecular Microbiology, Vol. 62, Issue 5 https://doi.org/10.1111/mmi.2006.62.issue-5	journal	December 2006
Raman microscopy for dynamic molecular imaging of living cells Hamada, Keisaku; Fujita, Katsumasa; Smith, Nicholas Isaac Journal of Biomedical Optics, Vol. 13, Issue 4 https://doi.org/10.1117/1.2952192	journal	January 2008
Bacterial Biofilms: A Common Cause of Persistent Infections Costerton, J. W. Science, Vol. 284, Issue 5418, p. 1318-1322 https://doi.org/10.1126/science.284.5418.1318	journal	May 1999
Role of Pseudomonas aeruginosa Peptidoglycan-Associated Outer Membrane Proteins in Vesicle Formation Wessel, Aimee K.; Liew, Jean; Kwon, Taejoon Journal of Bacteriology, Vol. 195, Issue 2 https://doi.org/10.1128/JB.01253-12	journal	January 2013
Pseudomonas aeruginosa Exotoxin Pyocyanin Causes Cystic Fibrosis Airway Pathogenesis Caldwell, Charles C.; Chen, Yi; Goetzmann, Holly S. The American Journal of Pathology, Vol. 175, Issue 6 https://doi.org/10.2353/ajpath.2009.090166	journal	December 2009

Similar Records

Pseudomonas aeruginosa forms Biofilms in Acute InfectionIndependent of Cell-to-Cell Signaling

Journal Article · Wed Sep 20 00:00:00 EDT 2006 · Infection and Immunity · OSTI ID:923645

Spatial Mapping of Pyocyanin in Pseudomonas Aeruginosa Bacterial Communities Using Surface Enhanced Raman Scattering

Journal Article · Tue Jun 28 00:00:00 EDT 2016 · Applied Spectroscopy · OSTI ID:1811273

Electrochemical Surface-Enhanced Raman Spectroscopy of Pyocyanin Secreted by Pseudomonas aeruginosa Communities

Journal Article · Wed May 01 00:00:00 EDT 2019 · Langmuir · OSTI ID:1765171

Related Subjects

59 BASIC BIOLOGICAL SCIENCES
Pseudomonas aeruginosa
Raman spectroscopy
biofilms
phenazines
quinolones

Label-free molecular imaging of bacterial communities of the opportunistic pathogen Pseudomonas aeruginosa

Citation Formats

References (26)

Similar Records

Related Subjects