Zang, YanyiRoberts, Teryn R.Harea, George T.Beely, Brendan M.Olivera Perez, Leonardo J.Ande, SreedeviBatchinsky, MariaLee, Ji H.Thrailkill, Marianne A.Reynolds, Melissa M.Batchinsky, Andriy I.2024-03-152024-03-152023-09-20Zang Y, Roberts TR, Harea GT, et al. Quantitative Analysis of Clot Deposition on Extracorporeal Life Support Membrane Oxygenators Using Digital and Scanning Electron Microscopy Imaging Techniques. Bio Protoc. 2023;13(18):e4814. Published 2023 Sep 20. doi:10.21769/BioProtoc.4814https://hdl.handle.net/1805/39266Device-induced thrombosis remains a major complication of extracorporeal life support (ECLS). To more thoroughly understand how blood components interact with the artificial surfaces of ECLS circuit components, assessment of clot deposition on these surfaces following clinical use is urgently needed. Scanning electron microscopy (SEM), which produces high-resolution images at nanoscale level, allows visualization and characterization of thrombotic deposits on ECLS circuitry. However, methodologies to increase the quantifiability of SEM analysis of ECLS circuit components have yet to be applied clinically. To address these issues, we developed a protocol to quantify clot deposition on ECLS membrane oxygenator gas transfer fiber sheets through digital and SEM imaging techniques. In this study, ECLS membrane oxygenator fiber sheets were obtained, fixed, and imaged after use. Following a standardized process, the percentage of clot deposition on both digital images and SEM images was quantified using ImageJ through blind reviews. The interrater reliability of quantitative analysis among reviewers was evaluated. Although this protocol focused on the analysis of ECLS membrane oxygenators, it is also adaptable to other components of the ECLS circuits such as catheters and tubing. Key features: • Quantitative analysis of clot deposition using digital and scanning electron microscopy (SEM) techniques • High-resolution images at nanoscale level • Extracorporeal life support (ECLS) devices • Membrane oxygenators • Blood-contacting surfacesen-USAttribution-NonCommercial 4.0 InternationalQuantitative analysisScanning electron microscopyMembrane oxygenatorExtracorporeal life support (ECLS)Clot depositionThrombosisArticle