荧光显微镜观察封闭式脊髓窗内的大鼠脊髓微循环

基础医学与临床 ›› 2013, Vol. 33 ›› Issue (8): 1019-1023.

荧光显微镜观察封闭式脊髓窗内的大鼠脊髓微循环

苑晓晨¹,²,李炳蔚²,武清斌³,荆瀛黎⁴,盛有明³,李宏伟²,刘淑英²,修瑞娟²

1.
2. 中国医学科学院北京协和医学院微循环研究所
3. 中国医学科学院&北京协和医学院微循环研究所
4. 中国医学科学院&北京协和医学院微循环研究所

收稿日期:2012-06-20 修回日期:2012-09-25 出版日期:2013-08-05 发布日期:2013-07-18
通讯作者: 修瑞娟 E-mail:xiurj@yahoo.com.cn
基金资助:
2011年公益性科研院所基本科研业务费专项基金

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

Received:2012-06-20 Revised:2012-09-25 Online:2013-08-05 Published:2013-07-18
Contact: Rui-juan XIU E-mail:xiurj@yahoo.com.cn

摘要/Abstract

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（5mg/L）体外孵育，静脉注射入体内；用罗丹明6G（0.3g/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粘附和流速，观察和记录罗丹明6G标记的白细胞在血管壁的粘附滚动。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。

关键词: 大鼠, 脊髓, 微循环, 活体荧光, 脊髓窗

Abstract: Objective To investigate the feasibility and efficacy of improved closed spinal window applied to assess the rat dorsal pial microcirculation by fluorescence microscopy. Methods On the 8-week-old SD rats, we made a precise improved closed spinal window with a laminectomy at the T10 level using a dental acrylic resin and a cover glass. The window was placed 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 100μL hematocrit red blood cells (BRC) by centrifugation (2000×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 microcirculation is recorded in real time through the window. Results Microvascular 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. Conclusion By improved closed spinal window and vertebral fixation device, we observed the microcirculation in the rat pia.

Key words: rat, spinal, microcirculation, intravital fluorescence, spinal window

中图分类号:

R744.9
R-332

苑晓晨李炳蔚武清斌荆瀛黎盛有明李宏伟刘淑英修瑞娟. 荧光显微镜观察封闭式脊髓窗内的大鼠脊髓微循环[J]. 基础医学与临床, 2013, 33(8): 1019-1023.

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