主动脉结扎灌注法在大鼠脑组织在体心脏灌流固定术中的应用

APPLICATION OF AORTIC LIGATION AND PERFUSION IN PERFUSION FIXATION OF LIVE RAT BRAIN TISSUES

目的探讨主动脉结扎灌注法在大鼠脑组织在体心脏灌流固定术中的应用效果。方法大鼠麻醉后,采用主动脉结扎灌注法依次灌注生理盐水和多聚甲醛固定脑组织。切取大鼠脑组织,进行尼氏和免疫组织染色。与传统的左心室进针灌注固定法比较,评价改进的主动脉结扎灌注法的灌注固定效果。结果主动脉结扎灌注法灌注固定成功率为(95.00±5.10)%,高于左心室进针灌注固定法的(80.83±6.29)%,差异有统计学意义(t=4.949,P<0.05)。两种方法灌注固定的脑组织均结构完整,尼氏染色和免疫荧光染色均显示,尼氏小体在核周围颗粒大,呈块状或颗粒状分布,清晰可辨,近边缘渐小而细,NeuN阳性细胞在组织片上清晰可见。结论主动脉结扎灌注法灌注固定成功率高,且可以取得与传统方法固定脑组织相同的良好效果。

Objective To investigate the effect of aortic ligation and perfusion in the perfusion-fixation of live rat brain tissues. Methods Rats were anesthetized and treated with aortic ligation.Rat brains were fixed by perfusion with normal saline solution,followed by paraformaldehyde.The fixed brains were then dissected and sliced for Nissl and immunohistochemical staining.The perfusion-fixation effect of aortic ligation and perfusion,as a modified perfusion-fixation method,was evaluated compared with the traditional left ventricular perfusion. Results The modified perfusion-fixation method had a significantly higher success rate than the traditional control method（（95.00±5.10）% vs（80.83±6.29）%;t=4.949,P0.05）.The brain structure was intact by both perfusion-fixation methods.Nissl and immunofluorescence staining showed that large massive or granular Nissl bodies were distributed surrounding the nucleus and clearly visible,as compared with those small and tiny bodies nearing the edge.Moreover,NeuN-positive cells were clearly observed in tissue slices. Conclusion The aortic ligation and perfusion can have a higher success rate and achieves the same brain fixation effect as the traditional method.