Cost-Effective and Versatile Analysis of Archaeal Surface Adhesion Under Shaking and Standing Conditions

Biofilms are aggregates of cells surrounded by an extracellular matrix providing protection from external stresses. While biofilms are commonly studied in bacteria, archaea also form such cell aggregates both in liquid cultures and on solid surfaces. Biofilm architectures vary when in liquid cultures versus on surfaces as well as when incubated under static conditions versus under shear forces of flowing liquid. Moreover, biofilms develop dynamically over time. Here, we describe surface adhesion assays employing a cost-effective, 3D-printed coverslip holder that can be used under a broad range of standing and shaking culture conditions. This multi-panel adhesion (mPAD) mount further allows the same culture to be sampled at multiple time points, ensuring consistency and comparability between samples and enabling analysis of the dynamics of biofilm formation. Additionally, a traditional surface adhesion assay in a 12-well plate under standing conditions is outlined as well. We anticipate the combination of these protocols to be useful for analyzing a wide array of biofilms and answering a multitude of biological questions.