Trantum Joshua R, Jayagopal Ashwath
Vanderbilt University, Nashville, TN, USA.
Methods Mol Biol. 2013;1026:35-44. doi: 10.1007/978-1-62703-468-5_3.
Atherosclerosis, a leading cause of morbidity and mortality worldwide, is characterized by the accumulation of lipid deposits inside arterial walls, leading to narrowing of the arterial lumen. A significant challenge in the development of diagnostic and therapeutic strategies is to elucidate the contribution of the various cellular participants, including macrophages, endothelial cells, and smooth muscle cells, in the initiation and progression of the atheroma. This protocol details a strategy using quantum dot nanocrystals to monitor homing and distribution of cell populations within atherosclerotic lesions with high signal to noise ratios over prolonged periods of analysis. This fluorescence-based approach enables the loading of quantum dots into cells such as macrophages without perturbing native cell functions in vivo, and has been used for the multiplexed imaging of quantum dot-labeled cells with biomarkers of atherosclerotic disease using conventional immunofluorescence techniques.
动脉粥样硬化是全球发病和死亡的主要原因,其特征是动脉壁内脂质沉积的积累,导致动脉管腔狭窄。在诊断和治疗策略的开发中,一个重大挑战是阐明包括巨噬细胞、内皮细胞和平滑肌细胞在内的各种细胞成分在动脉粥样瘤发生和发展中的作用。本方案详细介绍了一种使用量子点纳米晶体的策略,用于在长时间分析过程中以高信噪比监测动脉粥样硬化病变内细胞群体的归巢和分布。这种基于荧光的方法能够将量子点加载到巨噬细胞等细胞中,而不会在体内干扰天然细胞功能,并且已被用于使用传统免疫荧光技术对量子点标记细胞与动脉粥样硬化疾病生物标志物进行多重成像。