Rovainen C M, Woolsey T A, Blocher N C, Wang D B, Robinson O F
Department of Cell Biology, Washington University School of Medicine, St. Louis, MO 63110.
J Cereb Blood Flow Metab. 1993 May;13(3):359-71. doi: 10.1038/jcbfm.1993.49.
Cortical surface vessels were monitored through closed cranial windows with an epifluorescence microscope and SIT or ICCD cameras. Fluorescent dextrans or 1.3 microns latex beads were injected into the contralateral jugular vein for plasma labeling and for vascular transits. For close arterial transits, these tracers or physiological saline were injected into the ipsilateral external carotid artery. AVTTs were calculated from intensity differences of tracers between a branch of the MCA and a vein draining the same cortical region over time. AVTTs for saline dilutions of RBCs were significantly shorter (0.73 times) than for dextrans. Both dextrans and beads distributed with plasma. With FITC-dextran, inner diameters of arterioles and venules averaged 6 microns larger than hemoglobin under green light. This difference was likely due to the segregation of red blood cells and plasma during flow. Velocities of individual fluorescent beads were measured in pial vessels by strobe epi-illumination. Plots of bead velocities against radial position in arterioles were blunted parabolas. Peak shear rates in the marginal layer next to the vessel walls were determined directly from bead tracks in arterioles (D = 21-71 microns) and were 1.32 times the Poiseuille estimate. The calculated peak wall shear stress was 39 +/- 14 dyn/cm2 (mean +/- SD) for these arterioles but was probably severalfold greater in the smallest terminal pial arterioles. Vmax near the axes of arterioles increased with D+0.5. The calculated peak wall shear rate was highest in small arterioles and decreased with D-0.5. The calculated flow Q increased with D+2.5. These methods permit direct, simultaneous, dynamic measurements on multiple identified cerebral microvessels.
通过闭合颅骨窗,使用落射荧光显微镜以及慢扫描电荷耦合器件(SIT)或增强型电荷耦合器件(ICCD)相机监测皮质表面血管。将荧光葡聚糖或1.3微米的乳胶珠注入对侧颈静脉进行血浆标记和血管转运研究。对于近动脉转运,将这些示踪剂或生理盐水注入同侧颈外动脉。通过测量随时间变化的大脑中动脉(MCA)分支与引流相同皮质区域的静脉之间示踪剂的强度差异来计算动静脉转运时间(AVTT)。红细胞生理盐水稀释液的AVTT明显短于葡聚糖(为葡聚糖的0.73倍)。葡聚糖和乳胶珠均随血浆分布。使用异硫氰酸荧光素(FITC)标记的葡聚糖时,在绿光下小动脉和小静脉的内径平均比血红蛋白大6微米。这种差异可能是由于血流过程中红细胞和血浆的分离所致。通过频闪落射照明测量软脑膜血管中单个荧光珠的速度。小动脉中荧光珠速度相对于径向位置的曲线为钝抛物线。紧邻血管壁的边缘层中的峰值剪切速率直接从小动脉(直径D = 21 - 71微米)中的珠轨迹确定,是泊肃叶估计值的1.32倍。这些小动脉计算得出的峰值壁面剪切应力为39 +/- 14达因/平方厘米(平均值 +/- 标准差),但在最小的终末软脑膜小动脉中可能要大几倍。小动脉轴附近的最大速度(Vmax)随D + 0.5增加。计算得出的峰值壁面剪切速率在小动脉中最高,并随D - 0.5降低。计算得出的流量Q随D + 2.5增加。这些方法允许对多个已识别的脑微血管进行直接、同步、动态测量。