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In vivo near-infrared autofluorescence imaging of retinal pigment epithelial cells with 757 nm excitation

机译:757 nm激发的视网膜色素上皮细胞的体内近红外自发荧光成像

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摘要

We demonstrate near-infrared autofluorescence (NIRAF) imaging of retinal pigment epithelial (RPE) cells in vivo in healthy volunteers and patients using a 757 nm excitation source in adaptive optics scanning laser ophthalmoscopy (AOSLO). NIRAF excited at 757 nm and collected in an emission band from 778 to 810 nm produced a robust NIRAF signal, presumably arising from melanin, and revealed the typical hexagonal mosaic of RPE cells at most eccentricities imaged within the macula of normal eyes. Several patterns of altered NIRAF structure were seen in patients, including disruption of the NIRAF over a drusen, diffuse hyper NIRAF signal with loss of individual cell delineation in a case of non-neovascular age-related macular degeneration (AMD), and increased visibility of the RPE mosaic under an area showing loss of photoreceptors. In some participants, a superposed cone mosaic was clearly visible in the fluorescence channel at eccentricities between 2 and 6° from the fovea. This was reproducible in these participants and existed despite the use of emission filters with an optical attenuation density of 12 at the excitation wavelength, minimizing the possibility that this was due to bleed through of the excitation light. This cone signal may be a consequence of cone waveguiding on either the ingoing excitation light and/or the outgoing NIRAF emitted by fluorophores within the RPE and/or choroid and warrants further investigation. NIRAF imaging at 757 nm offers efficient signal excitation and detection, revealing structural alterations in retinal disease with good contrast and shows promise as a tool for monitoring future therapies at the level of single RPE cells.
机译:我们展示了健康志愿者和患者在自适应光学扫描激光检眼镜(AOSLO)中使用757 nm激发源的体内视网膜色素上皮(RPE)细胞的近红外自发荧光(NIRAF)成像。 NIRAF在757 nm处激发,并聚集在778至810 nm的发射带中,产生了很强的NIRAF信号,大概是黑色素产生的,并揭示了正常眼黄斑中成像的大多数偏心率下RPE细胞的典型六边形马赛克。在患者中观察到几种NIRAF结构改变的模式,包括在玻璃疣上的NIRAF破坏,在非血管性年龄相关性黄斑变性(AMD)情况下弥散性高NIRAF信号丧失单个细胞的轮廓,以及RPE马赛克在显示感光细胞丢失的区域下。在一些参与者中,在距中央凹2至6°的偏心距处,在荧光通道中清晰可见重叠的锥形镶嵌。这在这些参与者中是可重现的,尽管使用了在激发波长处光衰减密度为12的发射滤光片,但仍然存在,从而将这归因于激发光的泄漏的可能性降至最低。该圆锥信号可能是圆锥波在RPE和/或脉络膜内的荧光团发出的入射激发光和/或发出的NIRAF上导波的结果,需要进一步研究。 757 nm的NIRAF成像可提供有效的信号激发和检测,揭示视网膜疾病中的结构改变,具有良好的对比度,并有望作为监测单个RPE细胞水平未来疗法的工具。

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