Protocol abstract

Nature Protocols 5, 1236 - 1254 (2010)
Published online: 10 June 2010 | doi:10.1038/nprot.2010.71

Subject Categories: Cell and tissue culture | Microbiology and virology | Model organisms | Pharmacology and toxicology

Microtiter susceptibility testing of microbes growing on peg lids: a miniaturized biofilm model for high-throughput screening

Joe J Harrison1, Carol A Stremick2, Raymond J Turner2, Nick D Allan3, Merle E Olson3 & Howard Ceri2

Batch culture of biofilms on peg lids is a versatile method that can be used for microtiter determinations of biofilm antimicrobial susceptibility. In this paper, we describe a core protocol and a set of parameters (surface composition, the rate of rocking or orbital motion, temperature, cultivation time, inoculum size, atmospheric gases and nutritional medium) that can be adjusted to grow single- or multispecies biofilms on peg surfaces. Mature biofilms formed on peg lids can then be fitted into microtiter plates containing test agents. After a suitable exposure time, biofilm cells are disrupted into a recovery medium using sonication. Microbicidal endpoints can be determined qualitatively using optical density measurements or quantitatively using viable cell counting. Once equipment is calibrated and growth conditions are at an optimum, the procedure requires ~5 h of work over 4–6 d. This efficient method allows antimicrobial agents and exposure conditions to be tested against biofilms on a high-throughput scale.

  1. Department of Microbiology, University of Washington School of Medicine, Seattle, Washington, USA.
  2. Biofilm Research Group and Department of Biological Sciences, University of Calgary, Calgary, Alberta, Canada.
  3. Innovotech Incorporated, Edmonton, Alberta, Canada.

Correspondence to: Joe J Harrison1 e-mail:

Correspondence to: Howard Ceri2 e-mail: