Abstract
Bacterial coaggregation is the specific recognition and adhesion of genetically distinct bacteria, and it is central to multispecies biofilm formation. Here, we present a protocol integrating visual, turbidimetry, epifluorescence microscopy, and thermodynamic analysis to study coaggregation. We describe procedures for performing coaggregation assays and analyzing complementary data from different assays. This integrated workflow provides a robust and multidimensional framework for investigating bacterial coaggregation in different contexts. For complete details on the use and execution of this protocol, please refer to Afonso et al.(1)(,)(2).