明胶墨汁与荧光微球显示大鼠视网膜微血管灌流效果的对比研究

Comparative study between the improved ink perfusion and fluorescent microspheres in the detection of the rat retina microvessel perfusion

目的:比较明胶墨汁灌注与荧光微球灌注对大鼠视网膜微血管灌流情况的显示效果。方法:12只成年健康SD大鼠随机分成荧光微球组(6只)和明胶墨汁组(6只)。明胶墨汁组分两步经左心室灌注明胶墨汁40ml。荧光微球组经股静脉注射荧光微球0.5ml。视网膜行铺片和切片,观察荧光微球和改良墨汁灌注显示的微血管灌流情况;两组视网膜切片还进行NeuN、Parvalbumin和GFAP的免疫组化染色。结果:荧光微球零散分布于视网膜铺片周边部和中央部视的网膜血管中;不能显示铺片的血管轮廓和切片中的血管截面。荧光微球与NeuN、Parvalbumin或GFAP免疫组化双标不能在同一组织中准确显示视网膜血管与神经细胞的紧密空间关系。改良墨汁灌注能清晰显示大鼠视网膜铺片中周边部和中央部的血管网,以及切片中的血管截面,而且此方法与NeuN、Parvalbumin或GFAP免疫组化双标能在同一组织中准确显示视网膜血管与神经细胞的紧密空间关系。结论:改良明胶墨汁灌注在显示大鼠视网膜微血管灌流情况方面优于荧光微球,适宜于在视网膜疾病研究中广泛使用。

Objective： To compare the improved ink perfusion with fluorescent microspheres in the detection of the rat retina microvessel perfusion. Methods： 12 adult SD rats were randomly divided into fluorescent microspheres group （n=6） and modified ink perfused group （n=6）. The rats of the modified ink perfused group were infused with 40 ml of ink - gelatin. The rats of the latter group were injected with 0.5ml of fluorescent microspheres. Fluorescent microspheres and improved ink perfusion were used to show perfusion level of microvessel of whole-mount retina and retinal section. NeuN, parvalbumin and GFAP immunohistochemistry were made in the retinal section at the same time. Results： The fluorescent microspheres scattered in the vessels of central and peripheral retina in the whole mount. They can＇t show the outline of vessels in the whole-mount and the vessel sections. Double labeling between fluorescent microspheres and NeuN, parvalbumin or GFAP immunohistochemistry couldn＇t show precisely the close spatial relationship between the retinal vessels and retinal neurons or glial cells. In contrast, improved ink perfusion could clearly display the vascular net of central and peripheral retina in the whole mount and the vessel in the retinal sections. Double labeling between improved ink perfusion and NeuN, parvalbumin or GFAP immunohistochemistry could show precisely the close spatial relationship between the retinal vessels and retinal neurons or glial cells in the same retina area. Conclusion： Improved ink infusion is better than the fluorescent microspheres in the detection of rat retinal microvessel perfusion, which should be widely applied for the study of eye diseases.