Editorial on the Research Topic Methods in biofilms: 2022 Biofilms, complex communities of microorganisms adhering to surfaces, play a significant role in various fields, including healthcare, industry, and the environment (Flemming et al., 2016). As our understanding of biofilms expands, so does the need for advanced research methods and techniques to study their formation, composition, and physiological characteristics (Azeredo et al., 2017). This Research Topic aims to present a captivating collection of articles that highlight novel methodologies in biofilm research. Biofilms present researchers with a range of challenges and opportunities. The articles in this Research Topic address these challenges by focusing on the development and adaptation of cutting-edge methods for studying biofilms. By bridging the gap between different scientific disciplines, these contributions provide a comprehensive view of biofilm research. From investigating the dynamics of antibiotic resistance selection using microfluidic chips to the specific detection of multiple pathogens in co-infection and mixed biofilms, these articles offer valuable insights into the complex nature of biofilm formation and interactions. The Research Topic features articles that showcase innovative methodologies to study various aspects of biofilms. For instance, an article introduces a microfluidic chip for realtime monitoring of antibiotic resistance selection in bacterial biofilms (Tang et al.). This technology provides a valuable tool for understanding the dynamics of antibiotic resistance emergence and selection. Another article describes the development of a TaqMan duplex real-time PCR method for simultaneous detection of Streptococcus suis and Actinobacillus pleuropneumoniae in co-infection and mixed biofilms (Yi et al.). This method offers enhanced specificity and sensitivity for accurate quantification of these pathogens, facilitating disease prevention and control. Additionally, the Research Topic includes an upgraded repository of antimicrobial peptides (AMPs) for biofilm studies. This resource, B-AMP v2.0, provides a comprehensive collection of biofilm protein targets and AMPs, facilitating research into specific biofilm targets and anti-biofilm strategies (Ravichandran et al.). Furthermore, the utilization of fluorescence in situ hybridization (FISH) techniques is explored in another article, showcasing its applications for visualizing and quantifying microorganisms, genes, and metabolites within biofilms (Barbosa et al.). These advancements in FISH-based techniques offer valuable insights into biofilm structure Frontiers in Cellular and Infection Microbiology frontiersin.org 01