Yuan J, Melder R J, Jain R K, Munn L L
Massachusetts General Hospital, Boston, MA, USA.
Biotechniques. 2001 Feb;30(2):388-94. doi: 10.2144/01302rr02.
Physical interactions between circulating cells and the vascular wall play a central role in inflammation, metastasis, atherosclerosis, and therapeutic cell delivery. Unfortunately, traditional in vitro flow assays cannot be used to visualize the details of cell-surface interactions in blood flow because of inappropriate geometry and the poor penetration of light in erythrocyte solutions. To overcome these obstacles, we have developed an agarose-cast cylindrical vessel system to examine the profiles of cells interacting with surfaces under flow conditions. This design allows observation and quantification of cell deformation as cells adhere to surfaces under dynamic flow conditions without modifying the microscope or optical path. Furthermore, our flow system is uniquely suited for monitoring the profiles of adherent leukocytes deforming in response to erythrocyte suspension flow. We have used this flow system to study the role of erythrocytes in leukocyte-substrate interactions. Our results show that the cell deformation index (the ratio of the cell length to cell height) is higher in erythrocyte solutions compared to erythrocyte-free saline. This novel lateral view flow system provides a powerful technique for visualizing and quantifying the morphological changes of cells in contact with substrates exposed to shear stress.
循环细胞与血管壁之间的物理相互作用在炎症、转移、动脉粥样硬化和治疗性细胞递送中起着核心作用。不幸的是,传统的体外流动分析无法用于可视化血流中细胞表面相互作用的细节,因为其几何形状不合适且光在红细胞溶液中的穿透性较差。为了克服这些障碍,我们开发了一种琼脂糖浇铸的圆柱形血管系统,以检查在流动条件下与表面相互作用的细胞轮廓。这种设计允许在动态流动条件下细胞粘附到表面时观察和量化细胞变形,而无需修改显微镜或光路。此外,我们的流动系统特别适合监测粘附的白细胞在红细胞悬液流动作用下变形的情况。我们已使用该流动系统研究红细胞在白细胞与底物相互作用中的作用。我们的结果表明,与无红细胞的生理盐水相比,红细胞溶液中的细胞变形指数(细胞长度与细胞高度之比)更高。这种新颖的侧视图流动系统为可视化和量化与暴露于剪切应力的底物接触的细胞的形态变化提供了一种强大的技术。