Biodynamical processes, especially in system biology, that occur far apart in space may be highly correlated. To study such biodynamics, simultaneous imaging over a large span at high spatio-temporal resolutions is highly desired. For example, large-scale recording of neural network activities over various brain regions is indispensable in neuroscience. However, limited by the field-of-view (FoV) of conventional microscopes, simultaneous recording of laterally distant regions at high spatio-temporal resolutions is highly challenging. Here, we propose to extend the distance of simultaneous recording regions with a custom micro-mirror unit, taking advantage of the long working distance of the objective and spatio-temporal multiplexing. We demonstrate simultaneous dual-region two-photon imaging, spanning as large as 9 mm, which is 4 times larger than the nominal FoV of the objective. We verify the system performance in in vivo imaging of neural activities and vascular dilations, simultaneously, at two regions in mouse brains as well as in spinal cords, respectively. The adoption of our proposed scheme will promote the study of systematic biology, such as system neuroscience and system immunology.

中文翻译：

---

跨度超过 9 mm 的体内生物动力学同步双区域双光子成像

生物动力学过程，特别是在系统生物学中，在空间上相距很远发生的过程可能是高度相关的。为了研究这种生物动力学，非常需要以高时空分辨率进行大跨度的同步成像。例如，大规模记录不同大脑区域的神经网络活动在神经科学中是必不可少的。然而，受传统显微镜视场（FoV）的限制，以高时空分辨率同时记录横向遥远区域非常具有挑战性。在这里，我们建议利用定制微镜单元延长同时记录区域的距离，利用物镜和时空复用的长工作距离。我们展示了同步双区域双光子成像，跨度高达 9 毫米，比物镜标称视场大 4 倍。我们分别在小鼠大脑和脊髓的两个区域同时验证了系统在神经活动和血管扩张的体内成像中的性能。我们提出的方案的采用将促进系统生物学的研究，例如系统神经科学和系统免疫学。