한빛사논문
- 조회479
Jeongmoo Han 1*, Sunwon Kim 2,3*, Hyun Jung Kim 3, Hyeong Soo Nam 1, Min Woo Lee 4, Joon Woo Song 3, Jin Won Kim 3†, Hongki Yoo 1†
1Department of Mechanical Engineering, Korea Advanced Institute of Science and Technology, Daejeon, South Korea (J.H., H.S.N., H.Y.).
2Department of Cardiology, Korea University Ansan Hospital, South Korea (S.K.).
3Multimodal Imaging and Theranostic Lab, Cardiovascular Center, Korea University Guro Hospital, Seoul, South Korea (S.K., H.J.K., J.W.S., J.W.K.).
4Center for Intelligent and Interactive Robotics, Korea Institute of Science and Technology, Seoul, South Korea (M.W.L.).
*J. Han and S. Kim contributed equally.
†J.W. Kim and H. Yoo shared senior authorship.
Correspondence to: Hongki Yoo, PhD, Jin Won Kim, MD, PhD
Abstract
Background: Autofluorescence lifetime (AFL) imaging, a robust technique that enables label-free molecular investigation of biological tissues, is being introduced into the field of cardiovascular diagnostics. However, detailed AFL characteristics of coronary arteries remain elusive and there is a lack of methodology enabling such characterization.
Methods: We developed multispectral fluorescence lifetime imaging microscopy (FLIM) based on analog-mean-delay. Freshly sectioned coronary arteries and atheromas, harvested from 5 swine models, were imaged using FLIM and stained to label lipids, macrophages, collagen, and smooth muscle cells. The components were quantitated from digitized histological images and compared with the corresponding FLIM. Multispectral AFL parameters derived from 2 different spectral bands (390 nm and 450 nm) were analyzed.
Results: FLIM provided a wide field-of-view, high-resolution AFL imaging of frozen sections. Principal compositions of coronary arteries, such as tunica media, tunica adventitia, elastic laminas, smooth muscle cell-enriched fibrous plaque, lipid-rich core, and foamy macrophages, were well visualized in FLIM images and were found to have each different AFL spectra. In particular, proatherogenic components including lipids and foamy macrophages exhibited significantly different AFL values compared with plaque-stabilizing collagen- or smooth muscle cell-enriched tissues (P<0.0001). Pairwise comparisons showed that each composition was distinguishable from another by the difference in multispectral AFL parameters. Pixel-level analysis based on coregistered FLIM-histology dataset showed that each component of atherosclerosis (lipids, macrophages, collagen, and smooth muscle cells) had distinct correlation pattern with AFL parameters. Random forest regressors trained with the dataset allowed automated, simultaneous visualization of the key atherosclerotic components with high accuracy (r>0.87).
Conclusions: FLIM provided detailed pixel-level AFL investigation of the complex composition of coronary artery and atheroma. Our FLIM strategy enabling an automated, comprehensive visualization of multiple plaque components from unlabeled sections will be highly useful to efficiently evaluate ex vivo samples without the need for histological staining and analysis.
논문정보
- Research article
- 2023년 05월 (BRIC 등록일 2023-06-13)
- 국내 연구진
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