Quantitative Analysis of Clot Deposition on Extracorporeal Life Support Membrane Oxygenators Using Digital and Scanning Electron Microscopy Imaging Techniques

Yanyi Zang; Teryn R Roberts; George T Harea; Brendan M Beely; Leonardo Olivera J Perez; Sreedevi Ande; Maria Batchinsky; Ji H Lee; Marianne A Thrailkill; Melissa M Reynolds; Andriy I Batchinsky

doi:10.21769/BioProtoc.4814

Quantitative Analysis of Clot Deposition on Extracorporeal Life Support Membrane Oxygenators Using Digital and Scanning Electron Microscopy Imaging Techniques

Bio Protoc. 2023 Sep 20;13(18):e4814. doi: 10.21769/BioProtoc.4814.

Authors

Yanyi Zang^{1

2}, Teryn R Roberts^{1

2}, George T Harea^{1

2}, Brendan M Beely^{1

2}, Leonardo Olivera J Perez³, Sreedevi Ande⁴, Maria Batchinsky⁵, Ji H Lee⁶, Marianne A Thrailkill⁷, Melissa M Reynolds^{8

9}, Andriy I Batchinsky^{1

2}

Affiliations

¹ Autonomous Reanimation and Evacuation Research Program, The Geneva Foundation, San Antonio, TX, USA.
² Department of Translational Medicine, University of the Incarnate Word School of Osteopathic Medicine, San Antonio, TX, USA.
³ Indiana University School of Medicine, Indianapolis, IN, USA.
⁴ School of Mathematics, Science and Engineering, University of the Incarnate Word, San Antonio, TX, USA.
⁵ Texas Tech University Health Sciences Center School of Medicine, Lubbock, TX, USA.
⁶ Alaka'ina Foundation Family of Companies, Honolulu, HI, USA.
⁷ McGovern Medical School at The University of Texas Health Science Center at Houston, Houston, TX, USA.
⁸ Department of Chemistry and School of Biomedical Engineering, Colorado State University, Fort Collins, CO, USA.
⁹ School of Biomedical Engineering, Colorado State University, Fort Collins, CO, USA.

Abstract

Device-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 surfaces Graphical overview.

Keywords: Clot deposition; Extracorporeal life support (ECLS); ImageJ; Membrane oxygenator; Quantitative analysis; Scanning electron microscopy; Thrombosis.