Sepsis is a dysregulated response to infection that can lead to disseminated intravascular coagulation and life-threatening organ damage, with mortality rates as high as 40%. Polymorphonuclear leukocytes release webs (NETs) of decondensed chromatin decorated with histones and antimicrobial proteins that ensnare pathogens, degrade inflammatory cytokines, and reduce inflammation. However, when NETs are cleaved by DNases in the circulation, they release harmful degradation products including cell-free DNA, myeloperoxidase and histones that induce collateral tissue damage. Whether NETs are prothrombotic remains controversial in the literature, and which mechanisms underlie the interaction between NETs and pathogen, platelets, endothelial cells, and coagulation factors remain unclear. This Methods Collection aims to highlight recent advanced methods to investigate the role of NETs in the capture of bacteria, regulation of coagulation activity and platelet activation as well as NET-endothelium interaction in the setting of thrombosis and inflammation. The methods published in this collection will provide means to further our understanding of the contribution of NETs to the pathology of sepsis and other inflammatory diseases, and to assess the impact of various NET-targeted approaches as novel treatment for sepsis.