Label-free imaging of immune cell dynamics in the living retina using adaptive optics.
Label-free imaging of immune cell dynamics in the living retina using adaptive optics.
复制标题
使用自适应光学对活体视网膜中免疫细胞动力学的无标记成像。
DOI：
10.7554/elife.60547
复制
发表时间：
2020-10-14
期刊：
影响因子：
7.7
通讯作者：
Schallek J
中科院分区：
文献类型：
--
作者：
Joseph A;Chu CJ;Feng G;Dholakia K;Schallek J
我们最近的工作描述了视网膜血管系统中单个血细胞的运动(Joseph等人)。2019)使用自适应光学眼底镜。在这里，我们将这项技术应用于急性炎症的背景下，并发现在仅使用近红外光的情况下，在活的小鼠视网膜中既可以看到浸润性的免疫细胞，也可以看到组织驻留的免疫细胞，而没有任何标记。免疫细胞的活体成像可能受到手术操作、外源染料、转基因操作和光毒性的负面影响。这些混淆现在被克服了，使用相位对比度和延时视频成像来揭示髓系细胞在它们相互作用、渗出和观察小鼠视网膜时的动态行为。在同一视网膜位置，从炎症的开始到消退，在活体的几个小时到几个月的时间里，无创性地测量细胞的运动性和不同的血管反应。由于类似的系统已经可用于临床研究，这种方法很容易转化为人类应用。
Our recent work characterized the movement of single blood cells within the retinal vasculature (Joseph et al. 2019) using adaptive optics ophthalmoscopy. Here, we apply this technique to the context of acute inflammation and discover both infiltrating and tissue-resident immune cells to be visible without any labeling in the living mouse retina using near-infrared light alone. Intravital imaging of immune cells can be negatively impacted by surgical manipulation, exogenous dyes, transgenic manipulation and phototoxicity. These confounds are now overcome, using phase contrast and time-lapse videography to reveal the dynamic behavior of myeloid cells as they interact, extravasate and survey the mouse retina. Cellular motility and differential vascular responses were measured noninvasively and in vivo across hours to months at the same retinal location, from initiation to the resolution of inflammation. As comparable systems are already available for clinical research, this approach could be readily translated to human application.