Improved leukocyte tracking in mouse retinal and choroidal circulation
Improved leukocyte tracking in mouse retinal and choroidal circulation
复制标题
DOI：
10.1006/exer.2001.1134
复制
发表时间：
2002-03-01
影响因子：
3.4
通讯作者：
Crane, IJ
中科院分区：
文献类型：
--
作者：
Xu, HP;Manivannan, A;Crane, IJ
本研究的目的是发展一种新的方法，用它来显示白细胞在小鼠脉络膜和视网膜循环中的动力学。本研究中使用了有色（B10.RIII）和非有色（BALB/c）小鼠。通过小鼠尾静脉注射100穆尔0.05%荧光素钠以勾画血管轮廓，然后注射150穆尔（10（7）个细胞）C-AM标记的白细胞。用共焦激光扫描检眼镜获得眼底图像。动态图像序列同时记录在录像带（S-VIS）上，并以每秒25帧的速度进行数字化。这些数字图像随后用定制的基于个人计算机的图像分析系统进行分析。在荧光素血管造影术的最初几秒钟内，非色素小鼠的脉络膜和视网膜循环都可以可视化。然而，由于脉络膜中的快速荧光素渗漏，脉络膜和视网膜毛细血管循环的视图很快变得模糊。相比之下，在有色小鼠中，视网膜循环在脉络膜的黑暗背景下是清晰的，而脉络膜循环被色素上皮层掩盖，根本看不见。C-A-M标记的白细胞在所有实验小鼠的视网膜循环和非色素小鼠的脉络膜循环中清晰可见，长达30分钟。标记的循环细胞的数量随着时间的推移而减少。细胞在视网膜动脉中快速移动，在毛细血管系统中减慢甚至停止几秒钟，然后通过毛细血管后的小静脉和静脉再次稍微加快移动。在非色素小鼠中，观察到大量细胞在脉络膜循环中停滞。B10.RIII小鼠和BALB/c小鼠的血管直径、白细胞速度和切应力无差异。该方法允许白细胞的可视化，并提供关于它们在通过非色素小鼠的脉络膜和视网膜循环以及色素小鼠的视网膜循环时的行为的数据。它为生理条件下和眼部疾病发展过程中小鼠视网膜和脉络膜微循环中白细胞动态的真实的时间研究提供了一种有价值的新工具。(C)2002年爱思唯尔科学有限公司
The purpose of this study is to develop a now method with which to visualize leukocyte dynamics in murine choroidal and retinal circulation. Both pigmented (B10.RIII) and non-pigmented (BALB/c) mice were used in this study. One hundred mul of 0.05% sodium fluorescein was injected via the mice tail vein to outline the vessel, followed by 150 mul (10(7) cells) C-AM labelled leukocytes. Fundus images were obtained with a confocal scanning laser ophthalmoscope. The dynamic image sequences were recorded simultaneously on videotape (S-VIIS) and digitally at 25 frames per sec. The digital images were later analysed with a custom-made personal computer-based image analysis system. Both the choroidal and retinal circulation can be visualized in non-pigmented mice in the first few seconds of fluorescein angiography. However, the view of the choroidal and the retinal capillary circulation is soon blurred due to the rapid fluorescein leakage in the choroid. In contrast, in pigmented mice, retinal circulation is clear against the dark background of the choroid, while choroidal circulation is masked behind the pigment epithelial layer and cannot be seen at all. C-A-M labelled leucocytes were clearly seen in the retinal circulation of all experimental mice and in the choroidal circulation of non-pigmented mice for as long as 30 min. The number of labelled circulating cells decreased as time clasped. Cells moved rapidly in the retinal arteries, slowing down or even stopping for a few seconds in the capillary system, and then moved slightly faster again through the postcapillary venules and veins. In non-pigmented mice, significant number of cells were seen to have arrested in the choroidal circulation. There was no difference between B10.RIII mice and BALB/c mice in vessel diameters, leukocyte velocities and shear stresses. This method allows the visualization of leukocytes and provides data on their behavior as they move through the choroidal and retinal circulation of non-pigmented mice, and in the retinal circulation of pigmented mice. It provides a valuable new tool for the investigation of real time leukocyte dynamics in murine retinal and choroidal microcirculations both under physiological conditions and during the development of ocular disease. (C) 2002 Elsevier Science Ltd.