Label-free second harmonic generation imaging of cerebral vascular wall in local ischemia mouse model in vivo
Label-free second harmonic generation imaging of cerebral vascular wall in local ischemia mouse model in vivo
复制标题
DOI：
10.1016/j.neuroscience.2022.08.003
复制
发表时间：
2022-08
期刊：
影响因子：
3.3
通讯作者：
Hengfei Sun;Shu Wang;Jianxin Chen;Hongbo Yu
中科院分区：
文献类型：
--
作者：
Hengfei Sun;Shu Wang;Jianxin Chen;Hongbo Yu
二次谐波成像（second harmonic generation，SHG）具有无标记、无创等优点，已广泛应用于多种生物学和医学研究，但在活体脑组织中尚不多见。在本研究中，我们改进了经典的双光子激发荧光（TPEF）系统，在局部缺血小鼠模型中进行TPEF和SHG同时成像。在脑血管壁，我们发现了强SHG信号，它与胶原蛋白共定位。在连续2天的体内成像中，这种SHG信号在最初4 h内在局部缺血血管中保持稳定，然后在血栓形成后5 h其信号突然增加并在空间上增厚，这种趋势在随后的48 h内持续存在。本研究提供了直接和精确的时间轴的快速变化在脑血管壁在体内，并揭示了微妙的，但重要的时空动力学的结构信号在局部缺血。因此，这种脑蛋白活体SHG成像提供了一种强有力的工具来识别临床关键治疗时间窗周围的早期和细微的胶原病理变化。
Second harmonic generation (SHG) imaging is label-free and non-invasive, and it has been extensively applied in multiple biological and medical studies, but not in the brainin vivo. In this study, we modified classical two photon excited fluorescence (TPEF) system to performin vivosimultaneous TPEF and SHG imaging in the local ischemia mouse model. In cerebral vascular walls, we found strong SHG signal, which co-localized with collagen. In the continuous 2 days’in vivoimaging, this SHG signal remained stable in the local ischemic blood vessel in the initial 4 h, then its signal abruptly increased and got spatially thickened 5 h after thrombosis, and this tendency continued in the following 48 h. This study provides direct and precise timeline of rapid collagen change in cerebral vascular wallsin vivo, and reveals the subtle but significant temporal-spatial dynamics of this structural signal during local ischemia. Thus, this cerebralin vivoSHG imaging provides a powerful tool to identify the early and subtle pathological change of collagen around clinical key therapeutic time window.