Mempel Thorsten R, Moser Christian, Hutter Joerg, Kuebler Wolfgang M, Krombach Fritz
Institute for Surgical Research, University of Munich, Munich, Germany.
J Vasc Res. 2003 Sep-Oct;40(5):435-41. doi: 10.1159/000073902. Epub 2003 Oct 3.
Dynamic visualization of the intravascular events leading to the extravasation of leukocytes into tissues by intravital microscopy has significantly expanded our understanding of the underlying molecular processes. In contrast, the detailed observation of leukocyte transendothelial and interstitial migration in vivo has been hampered by the poor image contrast of cells within turbid media that is obtainable by conventional brightfield microscopy. Here we present a microscopic method, termed reflected light oblique transillumination microscopy, that makes use of the optical interference phenomena generated by oblique transillumination to visualize subtle gradients of refractive indices within tissues for enhanced image contrast. Using the mouse cremaster muscle, we demonstrate that this technique makes possible the reliable quantification of extravasated leukocytes as well as the characterization of morphological phenomena of leukocyte transendothelial and interstitial migration.
通过活体显微镜对导致白细胞渗入组织的血管内事件进行动态可视化，极大地扩展了我们对潜在分子过程的理解。相比之下，传统明场显微镜可获得的浑浊介质中细胞图像对比度较差，这阻碍了对体内白细胞跨内皮和间质迁移的详细观察。在此，我们提出一种称为反射光斜射透射显微镜的显微方法，该方法利用斜射透射产生的光学干涉现象来可视化组织内折射率的细微梯度，以增强图像对比度。利用小鼠提睾肌，我们证明该技术能够可靠地定量渗出的白细胞，并对白细胞跨内皮和间质迁移的形态学现象进行表征。
