Dirnagl U, Villringer A, Gebhardt R, Haberl R L, Schmiedek P, Einhäupl K M
Department of Neurology, University of Munich, Germany.
J Cereb Blood Flow Metab. 1991 May;11(3):353-60. doi: 10.1038/jcbfm.1991.74.
We used confocal laser scanning microscopy (CLSM) to investigate the morphology and three-dimensional relationships of the microcirculation of the superficial layers of the rat brain cortex in vivo. In anesthetized rats equipped with a closed cranial window (dura mater removed), after i.v. injection of 3 mg/100 g of body weight of fluorescein in 0.5 ml of saline, serial optical sections of the brain cortex intraparenchymal microcirculation were taken. Excitation was at a wavelength of 488 nm (argon laser), and emission was collected above 515 nm. CLSM provided images of brain vessels with sufficient signal-to-noise ratio for three-dimensional reconstructions down to a depth of 250 microns beneath the surface of the brain. Compared to conventional fluorescence microscopy, CLSM has a much higher axial resolution and higher depth of penetration. Laser light-induced intravascular aggregates, irregularities of erythrocyte flow, or microvascular occlusions ("light and dye injury") were not apparent in the current experimental paradigm. CLSM is a promising new tool for in vivo visualization of the cerebral microcirculation. Future studies have to characterize the potential damage to the tissue dye mechanisms.
我们使用共聚焦激光扫描显微镜（CLSM）在体内研究大鼠大脑皮层表层微循环的形态和三维关系。在配备闭合颅骨窗（硬脑膜已去除）的麻醉大鼠中，静脉注射0.5 ml生理盐水中含3 mg/100 g体重的荧光素后，获取大脑皮层实质内微循环的连续光学切片。激发波长为488 nm（氩激光），发射光在515 nm以上收集。CLSM提供了信噪比足够的脑血管图像，可用于三维重建，深度可达脑表面以下250微米。与传统荧光显微镜相比，CLSM具有更高的轴向分辨率和更深的穿透深度。在当前实验模式下，激光诱导的血管内聚集体、红细胞流动不规则或微血管阻塞（“光和染料损伤”）并不明显。CLSM是一种用于体内可视化脑微循环的有前景的新工具。未来的研究必须表征对组织染料机制的潜在损害。
