Lee Daniel M W, Zhang Min, Snyder Valerie C, Rossi Ethan A
University of Pittsburgh | Swanson School of Engineering, Department of Bioengineering, Pittsburgh, 15213, USA.
University of Pittsburgh | School of Medicine, Department of Ophthalmology, Pittsburgh, 15213, USA.
Sci Rep. 2024 Dec 3;14(1):30012. doi: 10.1038/s41598-024-81433-8.
The retinal pigment epithelium (RPE) is vital for the healthy function of the retina. Cellular level changes in the RPE are not visualized with current clinical techniques due to a lack of spatial resolution. Fluorescence adaptive optics scanning light ophthalmoscopy (AOSLO) can image RPE cells by utilizing their intrinsic autofluorescence (AF). The RPE AF has been imaged with only a few discrete excitation and emission bands and the multi-spectral AF has not been interrogated systematically at the level of single cells. In this study, we imaged 16 healthy eyes (ages 20-75) with AOSLO to investigate the multi-spectral AF as a function of age and wavelength with excitation from 650 - 805 nm. Quantitative analysis showed that 720 nm light produced images with the highest SNR (65.0 dB). Spatial AF variability showed a trend to increase with aging, suggesting increased heterogeneity in RPE AF with age. Spatial variability in the multi-spectral fluorescence of RPE cells with age may be a consequence of normal age-related loss of RPE cells. Multi-spectral fluorescence AOSLO provides new insight into aging related changes to RPE cells and may be a useful tool for studying diseases that affect the RPE, such as age-related macular degeneration (AMD).
视网膜色素上皮（RPE）对于视网膜的健康功能至关重要。由于缺乏空间分辨率，目前的临床技术无法观察到RPE细胞水平的变化。荧光自适应光学扫描激光检眼镜（AOSLO）可以利用RPE细胞的固有自发荧光（AF）对其进行成像。以往仅在少数离散的激发和发射波段对RPE的AF进行成像，尚未在单细胞水平系统地研究多光谱AF。在本研究中，我们使用AOSLO对16只健康眼睛（年龄20 - 75岁）进行成像，以研究激发波长在650 - 805 nm范围内多光谱AF随年龄和波长的变化情况。定量分析表明，720 nm光产生的图像具有最高的信噪比（65.0 dB）。空间AF变异性显示出随年龄增长而增加的趋势，表明RPE的AF异质性随年龄增加。RPE细胞多光谱荧光的空间变异性随年龄增加，可能是RPE细胞正常年龄相关丢失的结果。多光谱荧光AOSLO为RPE细胞与衰老相关的变化提供了新的见解，可能是研究影响RPE的疾病（如年龄相关性黄斑变性（AMD））的有用工具。
