McLeod D S, Lutty G A
Johns Hopkins University Applied Physics Laboratory, Laurel, Maryland.
Invest Ophthalmol Vis Sci. 1994 Oct;35(11):3799-811.
To develop a high-resolution histologic technique to study the postmortem human choroidal vasculature in dual perspective: vascular pattern in the flat perspective and structure in cross sections.
Fresh whole human choroids were denuded of retinal pigment epithelium, fixed, incubated for enzyme histochemical demonstration of alkaline phosphatase (APase) activity, bleached with hydrogen peroxide, and flat-embedded in glycol methacrylate. Vascular patterns were examined and documented en bloc, and subsequent serial sectioning was performed through specific sites of interest.
APase staining provided excellent visualization of the entire choroidal vasculature en bloc. Reaction product was generally more prominent in the choriocapillaris and collecting venules than in veins or arterioles, whereas arteries had the least activity. Diminished activity within focal regions of the choriocapillaris was observed in the far periphery of most aged subjects and was related to loss of endothelium and capillary atrophy. Hard drusen were generally observed in clusters located near collecting venules and appeared unrelated to any underlying angiopathy, whereas basal linear and laminar deposits were most often associated with regions of capillary dropout. Choroids from patients with diabetes demonstrated angiopathic changes consisting of extensive capillary dropout, beaded capillaries, neovascularization, and Bruch's membrane degeneration.
The benefits afforded by this method of analysis are that choroidal vasculature can be visualized in the native state and that nonvascular structures are retained for simultaneous analysis with vascular pattern and blood vessel structure.
开发一种高分辨率组织学技术,从两个视角研究死后人类脉络膜血管系统:平面视角下的血管模式和横截面结构。
将新鲜的完整人类脉络膜去除视网膜色素上皮,固定,进行酶组织化学以显示碱性磷酸酶(APase)活性,用过氧化氢漂白,然后平嵌入甲基丙烯酸乙二醇酯中。整体检查并记录血管模式,随后通过特定感兴趣部位进行连续切片。
APase染色能很好地整体显示整个脉络膜血管系统。反应产物在脉络膜毛细血管和集合小静脉中通常比在静脉或小动脉中更明显,而动脉的活性最低。在大多数老年受试者的最外周区域观察到脉络膜毛细血管局部区域的活性降低,这与内皮细胞丢失和毛细血管萎缩有关。硬性玻璃疣通常成簇出现在集合小静脉附近,似乎与任何潜在的血管病变无关,而基底线性和层状沉积物最常与毛细血管缺失区域相关。糖尿病患者的脉络膜表现出血管病变,包括广泛的毛细血管缺失、串珠状毛细血管、新生血管形成和布鲁赫膜变性。
这种分析方法的优点是可以在自然状态下观察脉络膜血管系统,并且保留非血管结构以便与血管模式和血管结构同时进行分析。