Harvesting and Disaggregation: An Overlooked Step in Biofilm Methods Research
Miller, Lindsey A.
Parker, Albert E.
Walker, Diane K.
Goeres, Darla M.
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Biofilm methods consist of four distinct steps: growing the biofilm in a relevant model, treating the mature biofilm, harvesting the biofilm from the surface and disaggregating the clumps, and analyzing the sample. Of the four steps, harvesting and disaggregation are the least studied but nonetheless critical when considering the potential for test bias. This article demonstrates commonly used harvesting and disaggregation techniques for biofilm grown on three different surfaces. The three biofilm harvesting and disaggregation techniques, gleaned from an extensive literature review, include vortexing and sonication, scraping and homogenization, and scraping, vortexing and sonication. Two surface types are considered: hard non-porous (polycarbonate and borosilicate glass) and porous (silicone). Additionally, we provide recommendations for the minimum information that should be included when reporting the harvesting technique followed and an accompanying method to check for bias.
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Buckingham-Meyer, K., Miller, L. A., Parker, A. E., Walker, D. K., Sturman, P., Novak, I., Goeres, D. M. Harvesting and Disaggregation: An Overlooked Step in Biofilm Methods Research. J. Vis. Exp. (182), e62390, doi:10.3791/62390 (2022).