Department of Ophthalmology, Fukushima Medical University School of Medicine, Fukushima City, Fukushima, Japan.
Invest Ophthalmol Vis Sci. 2011 Oct 31;52(11):8534-41. doi: 10.1167/iovs.11-8012.
The origin of autofluorescence in the subretinal space and the autofluorescence properties of the cells were investigated in surgically collected subretinal fluid.
Subretinal fluid was surgically collected from four eyes of patients with rhegmatogenous retinal detachment (three eyes) and Coats' disease (one eye). After cytocentrifuge preparation of the cells in the fluid and immunofluorescence staining, a cytologic examination was conducted by using confocal scanning laser microscopy. The autofluorescence of the cells was elucidated by measuring the fluorescence spectra with spectroscopy, to obtain different excitation laser light emission fingerprints.
The cells from the subretinal fluid were classified into three types: CD68-negative cells containing numerous pigmented granules, CD68-positive cells containing few pigments, and CD68-negative cells with no pigmented granules. Autofluorescence was observed in the inclusions of the cells classified into the former two types. When the cells were excited by a 458- or 488-nm laser light, emission spectra in autofluorescence showed little difference between CD68-positive and -negative cells. Peak analysis confirmed that the two types of cells showed the same emission peaks within this range of excitation light.
Autofluorescent inclusions appeared in the CD68-positive and -negative cells in the subretinal fluid. The macrophages in the subretinal fluid possess autofluorescence that is spectroscopically similar to lipofuscin. Autofluorescence of macrophages can be attributed to degenerated outer segments and debris from apoptotic photoreceptors. Clinicians should consider migration of macrophages, in addition to retinal pigment epithelium, as the possible source when abnormal fundus autofluorescence is observed using an ordinary set of fluorescence filters.
研究手术采集的视网膜下液中视网膜下空间自发荧光的起源和细胞的自发荧光特性。
从 4 只接受手术治疗的视网膜脱离（3 只眼）和 Coats 病（1 只眼）患者的眼内采集视网膜下液。对液体中的细胞进行细胞离心涂片和免疫荧光染色后，使用共聚焦扫描激光显微镜进行细胞学检查。通过光谱测量法对细胞的自发荧光进行解析，以获得不同激发激光的发射指纹。
视网膜下液中的细胞分为 3 种类型：CD68 阴性细胞，含有大量色素颗粒；CD68 阳性细胞，含少量色素；CD68 阴性细胞，无色素颗粒。在前两种类型的细胞中观察到内含物的自发荧光。当用 458nm 或 488nm 激光激发细胞时，自发荧光的发射光谱在 CD68 阳性和阴性细胞之间几乎没有差异。峰分析证实，这两种类型的细胞在该范围内的激发光下显示出相同的发射峰。
CD68 阳性和阴性的视网膜下液细胞中出现了自发荧光的内含物。视网膜下液中的巨噬细胞具有与脂褐素相似的光谱自发荧光。巨噬细胞的自发荧光归因于变性的外节和凋亡光感受器的碎片。当使用普通的荧光滤光片观察到异常的眼底自发荧光时，临床医生应考虑除了视网膜色素上皮细胞之外，巨噬细胞的迁移也是可能的来源。
