Label-free identification of human coronary atherosclerotic plaque based on a three-dimensional quantitative assessment of multiphoton microscopy images.

The rupture of coronary atherosclerotic plaque (CAP) and the resulting intracoronary thrombosis account for most acute coronary syndromes. Thus, the early identification and risk assessment of CAP is crucial for timely medical intervention. In this study, we propose a quantitative and label-free method for human CAP identification using multiphoton microscopy (MPM) and three-dimensional (3D) image analysis techniques. By detecting the intrinsic MPM signals, the microstructures of collagen and elastin fibers within normal and CAP-lesioned human coronary artery walls were imaged. Using a 3D gray level co-occurrence matrix method and 3D weighted vector summation algorithm, quantitative indicators that characterize the spatial texture and orientation features of the fibers were extracted. We demonstrate that these indicators show superior accuracy and repeatability over 2D texture features in CAP discrimination. Furthermore, by combining the 3D microstructural indicators, a support vector machine model that classifies CAP from the normal arterial wall with an accuracy of >97% was established. In conjunction with advances in multiphoton endoscopy, the proposed method shows great potential in providing a quantitative, label-free, and real-time tool for the early identification and risk assessment of CAP in the future.