Digital video-imaging of leukocyte migration in the iris: intravital microscopy in a physiological model during the onset of endotoxin-induced uveitis
Digital video-imaging of leukocyte migration in the iris: intravital microscopy in a physiological model during the onset of endotoxin-induced uveitis
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DOI：
10.1016/s0022-1759(00)00165-4
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发表时间：
2000-06-23
影响因子：
2.2
通讯作者：
Rosenbaum, JT
中科院分区：
文献类型：
--
作者：
Becker, MD;Nobiling, R;Rosenbaum, JT
炎症过程伴随着白细胞-内皮动力学的改变。内皮和白色细胞的相互变化允许白细胞放弃其正常的自由流动状态，以便滚动、停滞和迁移通过内皮。虽然活体显微镜是一种既定的方法来观察这一过程，眼睛一直未充分利用这一目的。虹膜脉管系统可以被视频摄影而没有创伤的伪影。采用罗丹明6 G（Rhodamine 6 G）对内毒素致炎模型的BALB/c小鼠白细胞进行活体染色，并采用数字视频技术分别记录内毒素致炎前、内毒素致炎后2 h、4 h的观察结果。微血流动力学参数的离线分析确定，表现出滚动的小静脉百分比从基线时的4%显著增加到内毒素注射后2小时的34%和4小时的82%。我们发现在4小时内白细胞停滞显著增加（601+/-119个细胞/mm 2，对照组动物为2+/-1个细胞/mm 2）。虽然剪切应力虹膜小动脉和小静脉之间的差异最小，滚动和逮捕优先发生在小静脉，表明剪切应力不是决定细胞粘附的主导因素。与以前的报告相比，活体显微镜，我们的方法包括细化或可视化细胞的优势，已经穿越以及避免手术创伤。该技术的分辨率和可量化性质使得该方法可应用于免疫应答期间白细胞-内皮动力学的重复观察。(C)2000 Elsevier Science B. V.保留所有权利。
The process of inflammation is accompanied by an alteration of leukocyte-endothelial dynamics. Reciprocal changes in the endothelium and the white cell permit the leukocyte to relinquish its normal free-flowing state in order to roll, arrest, and emigrate through the endothelium. Although intravital microscopy is an established method to observe this process, the eye has been under-utilized for this purpose. Iris vasculature can be videophotographed without the artifact of trauma. We used rhodamine 6G in vivo staining of leukocytes from BALB/c mice in a model of inflammation induced by intravitreally injected endotoxin, Digital video technology was used to record observations at baseline, 2 h, and 4 h after the endotoxin injection. Off-line analysis of microhemodynamic parameters established that the percentage of venules exhibiting rolling increased significantly from 4% at baseline to 34% at 2 h and 82% at 4 h after endotoxin injection. We found a marked increase in leukocyte arrest within 4 h (601+/-119 cells per mm(2) vs. 2+/-1 cells per mm2 in control animals). Although shear stress differs minimally between iris arterioles and venules, both rolling and arrest occurred preferentially in venules indicating that shear stress is not the dominant factor for determining cell adhesion. Compared to previous reports on intravital microscopy, our methodology includes refinements or advantages in visualizing cells that have transmigrated as well as the avoidance of surgical trauma. The resolution and quantifiable nature of this technique are such that the methodology can be applied to repetitive observation of leukocyte-endothelial dynamics during an immune response. (C) 2000 Elsevier Science B.V. All rights reserved.