Cideciyan Artur V, Swider Malgorzata, Jacobson Samuel G
Invest Ophthalmol Vis Sci. 2015 May;56(5):3393-406. doi: 10.1167/iovs.15-16726.
We previously developed reduced-illuminance autofluorescence imaging (RAFI) methods involving near-infrared (NIR) excitation to image melanin-based fluorophores and short-wavelength (SW) excitation to image lipofuscin-based flurophores. Here, we propose to normalize NIR-RAFI in order to increase the relative contribution of retinal pigment epithelium (RPE) fluorophores.
Retinal imaging was performed with a standard protocol holding system parameters invariant in healthy subjects and in patients. Normalized NIR-RAFI was derived by dividing NIR-RAFI signal by NIR reflectance point-by-point after image registration.
Regions of RPE atrophy in Stargardt disease, AMD, retinitis pigmentosa, choroideremia, and Leber congenital amaurosis as defined by low signal on SW-RAFI could correspond to a wide range of signal on NIR-RAFI depending on the contribution from the choroidal component. Retinal pigment epithelium atrophy tended to always correspond to high signal on NIR reflectance. Normalizing NIR-RAFI reduced the choroidal component of the signal in regions of atrophy. Quantitative evaluation of RPE atrophy area showed no significant differences between SW-RAFI and normalized NIR-RAFI.
Imaging of RPE atrophy using lipofuscin-based AF imaging has become the gold standard. However, this technique involves bright SW lights that are uncomfortable and may accelerate the rate of disease progression in vulnerable retinas. The NIR-RAFI method developed here is a melanin-based alternative that is not absorbed by opsins and bisretinoid moieties, and is comfortable to view. Further development of this method may result in a nonmydriatic and comfortable imaging method to quantify RPE atrophy extent and its expansion rate.
我们之前开发了低照度自发荧光成像(RAFI)方法,包括近红外(NIR)激发以成像基于黑色素的荧光团,以及短波(SW)激发以成像基于脂褐素的荧光团。在此,我们提议对NIR-RAFI进行归一化处理,以增加视网膜色素上皮(RPE)荧光团的相对贡献。
采用标准方案进行视网膜成像,保持健康受试者和患者的系统参数不变。在图像配准后,通过逐点将NIR-RAFI信号除以NIR反射率来获得归一化的NIR-RAFI。
根据脉络膜成分的贡献,由SW-RAFI上的低信号定义的Stargardt病、年龄相关性黄斑变性、色素性视网膜炎、脉络膜视网膜萎缩和Leber先天性黑蒙中的RPE萎缩区域,在NIR-RAFI上可能对应于广泛的信号范围。视网膜色素上皮萎缩往往总是对应于NIR反射率上的高信号。对NIR-RAFI进行归一化处理可减少萎缩区域信号的脉络膜成分。RPE萎缩面积的定量评估显示,SW-RAFI和归一化的NIR-RAFI之间无显著差异。
使用基于脂褐素的自发荧光成像对RPE萎缩进行成像已成为金标准。然而,该技术涉及明亮的SW光,令人不适,并且可能会加速脆弱视网膜疾病的进展速度。此处开发的NIR-RAFI方法是一种基于黑色素的替代方法,不会被视蛋白和双视黄醛部分吸收,观看舒适。该方法的进一步发展可能会产生一种无需散瞳且舒适的成像方法,用于量化RPE萎缩程度及其扩展速率。