Examination of Staphylococcus epidermidis Biofilms Using Flow-Cell Technology

  • Derek E. Moormeier
  • Kenneth W. Bayles
Part of the Methods in Molecular Biology book series (MIMB, volume 1106)


A common in vitro method to study Staphylococcus epidermidis biofilm development is to allow the bacteria to attach and grow on a solid surface in the presence of a continuous flow of nutrients. Under these conditions, the bacteria progress through a series of developmental steps, ultimately forming a multicellular structure containing differentiated cell populations. The observation of the biofilm at various time-points throughout this process provides a glimpse of the temporal changes that occur. Furthermore, use of metabolic stains and fluorescent reporters provides insight into the physiologic and transcriptional changes that occur within a developing biofilm. Currently, there are multiple systems available to assess biofilm development, each with advantages and disadvantages depending on the questions being asked. In this chapter, we describe the use of two separate flow-cell systems used to evaluate the developmental characteristics of staphylococcal biofilms: the FC270 flow-cell system from BioSurface Technologies, Corp. and the BioFlux1000 microfluidic flow-cell system from Fluxion Bioscience, Inc.

Key words

Biofilm BioFlux Flow-cell Epifluorescence microscopy Confocal laser scanning microscopy 


  1. 1.
    Costerton JW, Stewart PS, Greenberg EP (1999) Bacterial biofilms: a common cause of persistent infections. Science 284:1318–1322CrossRefPubMedGoogle Scholar
  2. 2.
    Clutterbuck AL, Woods EJ, Knottenbelt DC et al (2007) Biofilms and their relevance to veterinary medicine. Vet Microbiol 121:1–17CrossRefPubMedGoogle Scholar
  3. 3.
    Lewis K (2001) Riddle of biofilm resistance. Antimicrob Agents Chemother 45:999–1007CrossRefPubMedGoogle Scholar
  4. 4.
    Hornef MW, Wick MJ, Rhen M et al (2002) Bacterial strategies for overcoming host innate and adaptive immune responses. Nat Immunol 3:1033–1040CrossRefPubMedGoogle Scholar
  5. 5.
    Rogers KL, Fey PD, Rupp ME (2009) Coagulase-negative staphylococcal infections. Inf Dis Clin North Am 23:73–98CrossRefGoogle Scholar
  6. 6.
    Mann EE, Rice KC, Boles BR et al (2009) Modulation of eDNA release and degradation affects Staphylococcus aureus biofilm maturation. PLoS One 4:e5822CrossRefPubMedGoogle Scholar
  7. 7.
    Heydorn A, Nielsen AT, Hentzer M et al (2000) Quantification of biofilm structures by the novel computer program COMSTAT. Microbiology 146(Pt 10):2395–2407PubMedGoogle Scholar
  8. 8.
    Benoit MR, Conant CG, Ionescu-Zanetti C et al (2010) New device for high-throughput viability screening of flow biofilms. Appl Environ Microbiol 76:4136–4142CrossRefPubMedGoogle Scholar
  9. 9.
    Moormeier DE, Endres JL, Mann EE et al (2013) Use of microfluidic technology to analyze gene expression during Staphylococcus aureus biofilm formation reveals distinct physiological niches. Appl Environ Microbiol 79:3413–3424CrossRefPubMedGoogle Scholar

Copyright information

© Springer Science+Business Media, LLC 2014

Authors and Affiliations

  • Derek E. Moormeier
    • 1
  • Kenneth W. Bayles
    • 1
  1. 1.Department of Pathology and Microbiology, Center for Staphylococcal ResearchUniversity of Nebraska Medical CenterOmahaUSA

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