[Abstract]  [Full Text]  [PDF]

Joe J. Harrison², Howard Ceri², Jerome Yerly³, Carol A. Stremick², Yaoping Hu³, Robert Martinuzzi⁴ and Raymond J. Turner^1*

¹ Department of Biological Sciences, University of Calgary. T2N 1N4. Canada.
² Department of Biological Sciences and Biofilm Research Group, University of Calgary. T2N 1N4. Canada.
³ Electrical and Computer Engineering, Schulich School of Engineering, University of Calgary. T2N 1N4. Canada.
⁴ Mechanical and Manufacturing Engineering, Schulich School of Engineering, University of Calgary. 2500 University Drive N.W., Calgary, AB T2N 1N4. Canada.

* To whom correspondence should be addressed: Raymond J. Turner, Department of Biological Sciences, University of Calgary. T2N 1N4. Canada. Phone: 403-220-4308. Email: turnerr@ucalgary.ca

Biol. Proced. Online 2006;8:194-215. doi:10.1251/bpo127
Submitted: September 06, 2006; Accepted: October 28, 2006; Published: December 19, 2006.

Indexing terms: Biofilms; Imaging, Three-Dimensional.

Microbes frequently live within multicellular, solid surface-attached assemblages termed biofilms. These microbial communities have architectural features that contribute to population heterogeneity and consequently to emergent cell functions. Therefore, three-dimensional (3D) features of biofilm structure are important for understanding the physiology and ecology of these microbial systems. This paper details several protocols for scanning electron microscopy and confocal laser scanning microscopy (CLSM) of biofilms grown on polystyrene pegs in the Calgary Biofilm Device (CBD). Furthermore, a procedure is described for image processing of CLSM data stacks using amira^™, a virtual reality tool, to create surface and/or volume rendered 3D visualizations of biofilm microorganisms. The combination of microscopy with microbial cultivation in the CBD – an apparatus that was designed for high-throughput susceptibility testing – allows for structure-function analysis of biofilms under multivariate growth and exposure conditions.