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使用数字和扫描电子显微镜成像技术对体外膜肺氧合器上的血栓沉积进行定量分析。

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

作者信息

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

机构信息

Autonomous Reanimation and Evacuation Research Program, The Geneva Foundation, San Antonio, TX, USA.

Department of Translational Medicine, University of the Incarnate Word School of Osteopathic Medicine, San Antonio, TX, USA.

出版信息

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

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.

摘要

设备诱导的血栓形成仍然是体外生命支持(ECLS)的主要并发症。为了更全面地了解血液成分如何与ECLS回路组件的人工表面相互作用,迫切需要评估临床使用后这些表面上的凝块沉积情况。扫描电子显微镜(SEM)能够在纳米尺度上产生高分辨率图像,可用于观察和表征ECLS回路上的血栓沉积物。然而,提高ECLS回路组件SEM分析可量化性的方法尚未在临床上应用。为了解决这些问题,我们开发了一种通过数字和SEM成像技术对ECLS膜式氧合器气体传输纤维片上的凝块沉积进行量化的方案。在本研究中,获取使用后的ECLS膜式氧合器纤维片,进行固定和成像。按照标准化流程,通过ImageJ软件对数字图像和SEM图像上的凝块沉积百分比进行盲法评估。评估了审阅者之间定量分析的评分者间信度。尽管该方案侧重于ECLS膜式氧合器的分析,但它也适用于ECLS回路的其他组件,如导管和管道。关键特性 • 使用数字和扫描电子显微镜(SEM)技术对凝块沉积进行定量分析 • 纳米尺度的高分辨率图像 • 体外生命支持(ECLS)设备 • 膜式氧合器 • 血液接触表面 图形概述。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/cbd5/10518701/c45ac98eac4c/BioProtoc-13-18-4814-g001.jpg

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