Understanding how bacterial biofilms form and how to disrupt them requires effective methods for visualizing and modeling these complex communities of microorganisms. This collection aims to share significant advancements in imaging biofilm populations, tools or models for studying biofilm attachment to biologically relevant surfaces, and methods for demonstrating both in vitro and in vivo disruption of biofilms by new therapeutics. These advancements facilitate a better understanding of the intricate multicellular communities contained within biofilm matrices. Additionally, when exploring novel therapeutics that target biofilms, it is crucial to ensure that the biofilm model used is appropriate for accurately assessing biofilm eradication in vitro and in vivo. This collection seeks to promote transparency in the methodologies used in biofilm research and to expand the tools available for more rigorous biofilm studies.