Department of Biomedical Engineering, Texas A&M University, College Station, TX, USA.
Photochem Photobiol. 2010 May-Jun;86(3):727-31. doi: 10.1111/j.1751-1097.2010.00707.x. Epub 2010 Mar 10.
A prototype angioscopy system with fluorescence lifetime imaging microscopy (FLIM) capabilities was built and applied for biochemical imaging of human coronary atherosclerotic plaques. The FLIM angioscopy prototype consisted of a thin flexible angioscope suitable for UV-excited autofluorescence imaging, and a FLIM detection system based on a pulse sampling approach. The angioscope was composed of an imaging bundle attached to a gradient index objective lens and surrounded by a ring of illumination fibers (2 mm outer diameter, 50 microm spatial resolution). For FLIM detection based on the pulse sampling approach, a gated-intensified charge-couple device camera (200 ps temporal resolution) was used. Autofluorescence was excited with a pulsed UV laser (337 nm) and FLIM images were acquired at three emission bands (390/40 nm, 450/40 nm, 550/88 nm). The system was characterized on standard fluorophores and then used to image postmortem human coronary arteries. The FLIM angioscope allowed us to distinguish elastin-dominant plaques (peak emission at 450 nm, approximately 1.5 ns lifetimes) from collagen-dominant plaques (peak emission at 390 n, approximately 2-3 ns lifetimes) based on their intrinsic fluorescence spectral and lifetime differences. This study demonstrates the potential of FLIM angioscopy for biochemical imaging of human coronary atherosclerotic plaques.
一个具有荧光寿命成像显微镜(FLIM)功能的血管镜原型系统已经建立,并应用于人类冠状动脉粥样硬化斑块的生化成像。FLIM 血管镜原型系统由一个适合紫外激发自发荧光成像的薄型柔性血管镜和一个基于脉冲采样方法的 FLIM 检测系统组成。血管镜由一个成像束组成,连接到梯度折射率物镜,并被一圈照明光纤(外径 2 毫米,空间分辨率 50 微米)包围。对于基于脉冲采样方法的 FLIM 检测,使用了门控增强电荷耦合器件相机(200 皮秒时间分辨率)。自发荧光用脉冲紫外激光(337nm)激发,在三个发射带(390/40nm、450/40nm、550/88nm)获取 FLIM 图像。该系统在标准荧光团上进行了表征,然后用于成像死后的人类冠状动脉。FLIM 血管镜使我们能够根据其固有荧光光谱和寿命差异,区分富含弹性蛋白的斑块(发射峰在 450nm 左右,寿命约为 1.5ns)和富含胶原蛋白的斑块(发射峰在 390nm 左右,寿命约为 2-3ns)。这项研究表明了 FLIM 血管镜在人类冠状动脉粥样硬化斑块生化成像中的潜力。