荧光显微镜观察封闭式脊髓窗内的大鼠脊髓微循环  

Observation of the spinal microcirculation in rat through a closed spinal window by fluorescence microscopy

作　　者：苑晓晨[1] 李炳蔚[1] 武清斌[1] 荆瀛黎[1] 盛有明[1] 李宏伟[1] 刘淑英[1] 修瑞娟[1] 

机构地区：[1]中国医学科学院北京协和医学院微循环研究所,北京100005

出　　处：《基础医学与临床》2013年第8期1019-1023,共5页Basic and Clinical Medicine

基　　金：2011年中国医学科学院微循环研究所公益性科研院所基本科研业务费专项基金

摘　　要：目的探讨应用荧光显微镜通过改进的封闭式脊髓窗,观察大鼠脊髓背侧软脊膜微循环状态的可行性及效果。方法用8周龄雄性SD大鼠,暴露T9至T11棘突,咬除T10棘突和椎板,暴露硬脊膜;在T9和T11棘突上安装固定器辅助物。用牙科丙烯酸树脂和玻璃片为原料在T10节段安装改进的封闭脊髓窗。采血并离心(2 000×g)收集红细胞,将100μL压积的红细胞用Dil荧光染料(5 mg/L)体外孵育,静脉注射入体内;用罗丹明6G(0.3 g/L)静脉注射,体内标记白细胞。大鼠脊柱用微循环观测椎体固定器进行固定。用动物活体荧光成像显微系统进行观测和记录。结果通过该系统可以观测大鼠软脊膜上微血管的形态和分布特点、血管密度;可以观测到Dil标记的红细胞在血管内的黏附和流速,观察和记录罗丹明6G标记的白细胞在血管壁的黏附滚动。结论改进式的封闭脊髓窗和微循环观测椎体固定器,可用于对大鼠软脊膜上微血管进行观察和检测。Objective To investigate the feasibility and efficacy of closed spinal window applied to assess the rat dorsal pial mierocirculation by fluorescence microscopy. Methods On the 8-week-old SD rats, we made a precise improved closed spinal window with a laminectomy at the TIO level using a dental acrylic resin and a cover glass. The window was plaeed over the back of the rat with intact dura. Pasted the dental acrylic resin onto the spinous processes at the T9 and T11 as fixed aids. Collect 100 μL hematocrit red blood cells （BRC） by eentrifugation （2 000 ×g）. The RBC labelled by Dil （5 mg/L） in vitro were injected in jugular vein. Leukocytes labelled with rhodamine 6G were injected intravenously （0.3 g/L）. The spine was fixed with vertebral fixation device. Local mi- crocirculation is recorded in real time through the window. Results Mierovascular morphology, distribution and blood vessel density can be observed. Dil-labeled RBC velocity and adhesion were observed. Leukocyte adherence to pial vessels and assessment of perfusion were observed with the help of rhodamine 6G. Conclusions By im- proved closed spinal window and vertebral fixation device, we observed the microcirculation in the rat pia.

关 键 词：大鼠 脊髓 微循环 活体荧光 脊髓窗 

分 类 号：Q954[生物学—动物学]