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Detection and imaging of quorum sensing in Pseudomonas aeruginosa biofilm communities by surface-enhanced resonance Raman scattering

Nature Materials volume 15, pages 1203–1211 (2016)Cite this article

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Abstract

Most bacteria in nature exist as biofilms, which support intercellular signalling processes such as quorum sensing (QS), a cell-to-cell communication mechanism that allows bacteria to monitor and respond to cell density and changes in the environment. As QS and biofilms are involved in the ability of bacteria to cause disease, there is a need for the development of methods for the non-invasive analysis of QS in natural bacterial populations. Here, by using surface-enhanced resonance Raman scattering spectroscopy, we report rationally designed nanostructured plasmonic substrates for the in situ, label-free detection of a QS signalling metabolite in growing Pseudomonas aeruginosa biofilms and microcolonies. The in situ, non-invasive plasmonic imaging of QS in biofilms provides a powerful analytical approach for studying intercellular communication on the basis of secreted molecules as signals.

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Figure 1: Schematic representation of nanostructured porous substrates for in situ plasmonic detection and imaging of QS in Pseudomonas aeruginosa biofilms.
Figure 2: Raman and SERRS spectra of pyocyanin.
Figure 3: SERRS analysis of phenazine production by P. aeruginosa PA14 strains grown in planktonic cultures, recorded with Au@pNIPAM hydrogels.
Figure 4: In situ detection of pyocyanin secreted by P. aeruginosa PA14 colony-biofilms grown on Au@pNIPAM hydrogels.
Figure 5: In situ detection and imaging of pyocyanin produced by P. aeruginosa PA14 biofilms grown on mesostructured Au@TiO2 thin films.
Figure 6: In situ detection and imaging of pyocyanin produced by P. aeruginosa PA14 grown on micropatterned Au@SiO2 substrates.

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Acknowledgements

This work has been funded by the European Research Council (ERC Advanced Grant no. 267867 Plasmaquo). We thank D. K. Newman, Department of Biology, California Institute of Technology and Howard Hughes Medical Institute for providing us with P. aeruginosa PA14 and PA14 Δphz1/2 strains. We thank F. M. Ausubel, Department of Genetics, Harvard Medical School for providing P. aeruginosa PA14 phzH, phzM and phzS strains. E. Modin and A. Chuvilin are thanked for their work on FIB/SEM of supercrystals. E.H.H. gratefully acknowledges the Spanish Ministry of Economy and Competitiveness for funding a Juan de la Cierva Fellowship. V.M.-G. acknowledges an FPU scholarship from the Spanish MINECO.

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Authors and Affiliations

  1. Departamento de Química Física, Universidade de Vigo, 36310 Vigo, Spain

    Gustavo Bodelón, Verónica Montes-García, Vanesa López-Puente, Sergio Rodal-Cedeira, Celina Costas, Sirin Celiksoy, Ignacio Pérez-Juste, Jorge Pérez-Juste, Isabel Pastoriza-Santos & Luis M. Liz-Marzán

  2. Bionanoplasmonics Laboratory, CIC biomaGUNE, Paseo de Miramón 182, 20009 Donostia-San Sebastián, Spain

    Eric H. Hill, Cyrille Hamon, Marta N. Sanz-Ortiz, Leonardo Scarabelli, Andrea La Porta & Luis M. Liz-Marzán

  3. Ikerbasque, Basque Foundation for Science, 48013 Bilbao, Spain

    Luis M. Liz-Marzán

  4. Biomedical Research Networking Center in Bioengineering, Biomaterials, and Nanomedicine (CIBER-BBN), 20009 Donostia - San Sebastián, Spain

    Luis M. Liz-Marzán

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  1. Gustavo Bodelón
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Contributions

G.B. designed and executed bacterial growth experiments, V.M.-G. performed SERS experiments with Au@pNIPAM substrates, V.L.-P. fabricated Au@TiO2 substrates and measured SERS, C.C. executed bacterial growth experiments, S.R.-C. and S.C. participated in the fabrication of Au@pNIPAM substrates, I.P.-J. carried out DFT calculations, E.H.H. executed bacterial growth experiments and measured SERS on Au@SiO2 substrates, C.H., M.N.S.-O. and L.S. participated in the fabrication of Au@SiO2 substrates, A.L.P. carried out SEM imaging, I.P.-S. and J.P.-J. contributed to substrate design and discussion, L.M.L.-M. designed and supervised the project and the discussions. All authors participated in writing of the manuscript.

Corresponding authors

Correspondence to Gustavo Bodelón or Luis M. Liz-Marzán.

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Bodelón, G., Montes-García, V., López-Puente, V. et al. Detection and imaging of quorum sensing in Pseudomonas aeruginosa biofilm communities by surface-enhanced resonance Raman scattering. Nature Mater 15, 1203–1211 (2016). https://doi.org/10.1038/nmat4720

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