Imaging and quantitative analysis of atherosclerotic lesions by CARS-based multimodal nonlinear optical microscopy.
Imaging and quantitative analysis of atherosclerotic lesions by CARS-based multimodal nonlinear optical microscopy.
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DOI：
10.1161/atvbaha.109.189316
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发表时间：
2009-09
期刊：
影响因子：
--
通讯作者：
Cheng JX
中科院分区：
文献类型：
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作者：
Wang HW;Langohr IM;Sturek M;Cheng JX
根据AHA委员会建议的原始方案，评估无标记多模非线性光学(NLO)显微镜表征不同动脉粥样硬化病变类型的能力，从而实现对不同动脉粥样硬化病变类型的原位定量分析。取自24头雄性Ossabaw猪，分为瘦肉型对照组和代谢综合征组，用多模非线性光学显微镜成像，并在相干反斯托克斯拉曼散射(CARS)显微镜平台上集成和频产生(SFG)和双光子激发荧光(TPEF)。泡沫细胞、脂肪沉积、基质和纤维帽以亚微米三维分辨率显示。从早期的适应性内膜增厚到晚期的纤维性动脉粥样硬化或矿化，病变在无标记的情况下可见。每一个病变的组织学染色证实了病变的分期。根据CARS和SFG信号对脂质和胶原含量进行定量分析。增厚的内膜中脂质堆积在IV型病变中达到顶峰，而胶原沉积在V型病变中最高。腔内CARS成像显示有能力观察表面泡沫细胞的位置，这表明病变动脉中相对活跃的部位。我们已经证明了基于CARS的多模式NLO显微镜能够用亚细胞细节询问病变发展的不同阶段，从而能够对脂质和胶原含量进行定量分析。
Assess the ability of label-free multimodal nonlinear optical (NLO) microscopy to characterize, and thus enable quantitative in situ analyses of, different atherosclerotic lesion types, according to the original scheme suggested by the AHA Committee. Iliac arteries were taken from 24 male Ossabaw pigs divided into lean control and metabolic syndrome groups and were imaged by multimodal NLO microscopy where sum-frequency generation (SFG) and two-photon excitation fluorescence (TPEF) were integrated on a coherent anti-Stokes Raman scattering (CARS) microscope platform. Foam cells, lipid deposits, matrices, and fibrous caps were visualized with submicron 3-D resolution. Starting from the adaptive intimal thickening in the initial stage to the fibrous atheroma or mineralization in the advanced stages, lesions were visualized without labels. Histological staining of each lesion confirmed the lesion stages. Lipid and collagen contents were quantitatively analyzed based on the CARS and SFG signals. Lipid accumulation in thickened intima culminated in type IV while the highest collagen deposition was found in Type V lesions. Luminal CARS imaging showed the capability of viewing the location of superficial foam cells that indicate relatively active locus in a lesion artery. We have demonstrated the capability of CARS-based multimodal NLO microscopy to interrogate different stages of lesion development with subcellular detail to permit quantitative analysis of lipid and collagen contents.