眼底视网膜色素上皮层细胞脂褐素及氧化黑色素自体荧光寿命成像研究

Autofluorescence lifetime imaging of lipofuscin and oxidizedmelanin in retinal pigment epithelium cells

利用一种基于时间相关单光子计数器的双光子激发荧光寿命显微成像技术,对猪眼底视网膜色素上皮层细胞内的脂褐素和氧化黑色素颗粒的空间分布及其荧光寿命特性进行了研究,尤其对于这些色素颗粒在光致氧化环境中的荧光寿命差异进行了分析.结果表明,利用荧光寿命测量能有效区分视网膜色素上皮层细胞中的多组分荧光团,利用荧光寿命的衰减参数可分辨正常及异常的荧光现象.该方法有望发展成为一种用于眼科临床诊断及病理学研究的高灵敏度的工具,对眼底细胞随年龄增长的衰老机理的研究具有重要的意义.

We report on our investigation of the spatial distribution and autofluorescence lifetime characterization of lipofuscin and oxidized melanin in the retinal pigment epithelium cells of the pig eye using a two-photon excitation fluorescence lifetime imaging microscopy （TPE-FLIM） system, which that is based on a time-correlated single photon counting technique. In particular, we analyzed the difference of autofluorescence lifetimes of these pigment granules in light-induced oxidizing environment. The experimental results showed that the fluorescence lifetime imaging can provide an effective differentiation of multi-component fluorophores, and fluorescence decay can be used to distinguish normal from abnormal fluorescence. TPE-FLIM has the potential to provide a high sensitive imaging instrument for the clinical diagnosis and pathological studies in ophthalmology, and is also of significance to the study of aging mechanism of cells in the fundus.