Dirnagl U, Villringer A, Einhäupl K M
Department of Neurology, University of Munich, Germany.
J Microsc. 1992 Jan;165(Pt 1):147-57. doi: 10.1111/j.1365-2818.1992.tb04312.x.
Confocal scanning laser microscopy (CSLM) was used to study the microcirculation of the brain neocortex in anaesthetized rats. After removal of the dura mater, implantation of a closed cranial window, and intravenous injection of fluorescein, three-dimensional reconstructions of cortical capillaries were performed down to a depth of 250 microns below the pial surface. Using a one-dimensional approach (single line scanning), erythrocyte (negative contrast in fluorescently labelled plasma) and leucocyte (labelled with rhodamine 6 G) velocity and supply rate in cortical capillaries were measured. The effect of CO2-inhalation on capillary blood flow dynamics was studied. Capillaries were imaged continuously for up to 1 h without changes in flow or fluorescence pattern. However, by increasing the laser power 10-100-fold, aggregate formation was induced and capillaries were occluded, possibly due to damage to vascular endothelium. We conclude that CSLM can be used to study morphological and dynamic aspects of fluorescently labelled subsurface structures in organs of experimental animals.
共聚焦扫描激光显微镜（CSLM）用于研究麻醉大鼠大脑新皮质的微循环。去除硬脑膜、植入封闭的颅骨视窗并静脉注射荧光素后，对软脑膜表面以下250微米深度的皮质毛细血管进行三维重建。采用一维方法（单线扫描），测量皮质毛细血管中红细胞（荧光标记血浆中的负性对比）和白细胞（用罗丹明6G标记）的流速和供应率。研究了吸入二氧化碳对毛细血管血流动力学的影响。在血流或荧光模式无变化的情况下，对毛细血管连续成像长达1小时。然而，将激光功率提高10 - 100倍会诱导聚集体形成并阻塞毛细血管，这可能是由于血管内皮损伤所致。我们得出结论，CSLM可用于研究实验动物器官中荧光标记的地下结构的形态和动态方面。
