Visualizing Chemoattraction of Planktonic Cells to a Biofilm

  • Sneha Jani
Part of the Methods in Molecular Biology book series (MIMB, volume 1729)


Bacterial chemotaxis in response to continuous chemical gradients has been extensively studied at the individual cell and population levels using a variety of well-established in vitro methods (Englert et al., Microfluidic techniques for the analysis of bacterial chemotaxis. Methods Mol Biol 571:1–23, 2009). In nature, bacteria are surrounded by heterogeneous chemical gradients; hence, it is essential to understand chemotaxis behavior under such conditions. Here, we describe a setup that allows visualization of the chemotaxis response of motile cells to the complex microenvironment of a biofilm maintained under static conditions. The biofilm is separated from the motile cells by a semi-permeable membrane. Cells swimming toward the biofilm are captured on the membrane and imaged using confocal laser scanning microscopy (CLSM). Chemotaxis toward specific molecules produced by the biofilm, such as autoinducer-2 (AI-2), can be studied using this setup. This system can be adapted to study chemotaxis toward poly-species biofilms, or even mammalian cells.


Biofilm Chemotaxis Polycarbonate membrane Insert-in-well Autoinducer-2 Confocal laser scanning microscopy 


  1. 1.
    Costerton JW (1995) Overview of microbial biofilms. J Ind Microbiol 15:137–140CrossRefGoogle Scholar
  2. 2.
    Donlan RM (2002) Biofilms: microbial life on surfaces. Emerg Infect Dis 8:881–890CrossRefPubMedPubMedCentralGoogle Scholar
  3. 3.
    Davey ME, O'Toole GA (2000) Microbial biofilms: from ecology to molecular genetics. Microbiol Mol Biol Rev 64:847–867CrossRefPubMedPubMedCentralGoogle Scholar
  4. 4.
    Hall-Stoodley L, Costerton JW, Stoodley P (2004) Bacterial biofilms: from the natural environment to infectious diseases. Nat Rev Microbiol 2:95–108CrossRefPubMedGoogle Scholar
  5. 5.
    Parkinson JS, Houts SE (1982) Isolation and behavior of Escherichia coli deletion mutants lacking chemotaxis functions. J Bacteriol 151:106–113PubMedPubMedCentralGoogle Scholar
  6. 6.
    Hansen MC, Palmer RJ Jr, Udsen C, White DC, Molin S (2001) Assessment of GFP fluorescence in cells of Streptococcus gordonii under conditions of low pH and low oxygen concentration. Microbiology 147(Pt 5):1383–1391CrossRefPubMedPubMedCentralGoogle Scholar
  7. 7.
    Schneider CA, Rasband WS, Eliceiri KW (2012) NIH image to imageJ: 25 years of image analysis. Nat Methods 9:671–675CrossRefPubMedPubMedCentralGoogle Scholar
  8. 8.
    Reisner A, Krogfelt KA, Klein BM, Zechner EL, Molin S (2006) In vitro biofilm formation of commensal and pathogenic Escherichia coli strains: impact of environmental and genetic factors. J Bacteriol 188:3572–3581CrossRefPubMedPubMedCentralGoogle Scholar

Copyright information

© Springer Science+Business Media, LLC 2018

Authors and Affiliations

  1. 1.Molecular FoundryLawrence Berkeley National LaboratoryBerkeleyUSA

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