Department of Anesthesiology, The First Medical Center, Chinese PLA General Hospital, Beijing 100853, China.
Department of Automation, Tsinghua University, Beijing 100084, China.
Biosensors (Basel). 2022 Jul 26;12(8):567. doi: 10.3390/bios12080567.
The fluorescence microscope has been widely used to explore dynamic processes in vivo in mouse brains, with advantages of a large field-of-view and high spatiotemporal resolution. However, owing to background light and tissue scattering, the single-photon wide-field microscope fails to record dynamic neural activities in the deep brain. To achieve simultaneous imaging of deep-brain regions and the superficial cortex, we combined the extended-field-of-view microscopy previously proposed with a novel prism-based cranial window to provide a longitudinal view. As well as a right-angle microprism for imaging above 1 mm, we also designed a new rectangular-trapezoidal microprism cranial window to extend the depth of observation to 1.5 mm and to reduce brain injury. We validated our method with structural imaging of microglia cells in the superficial cortex and deep-brain regions. We also recorded neuronal activity from the mouse brains in awake and anesthesitized states. The results highlight the great potential of our methods for simultaneous dynamic imaging in the superficial and deep layers of mouse brains.
荧光显微镜已被广泛用于探索活体小鼠大脑中的动态过程，具有大视场和高时空分辨率的优点。然而，由于背景光和组织散射，单光子宽场显微镜无法记录深部大脑中的动态神经活动。为了实现深部脑区和浅层皮层的同时成像，我们将先前提出的扩展视场显微镜与新型棱镜式颅窗相结合，提供纵向视野。除了用于 1mm 以上成像的直角微棱镜外，我们还设计了一种新的矩形梯形微棱镜颅窗，将观察深度扩展到 1.5mm，并减少脑损伤。我们使用浅层皮层和深部脑区的小胶质细胞结构成像验证了我们的方法，还在清醒和麻醉状态下记录了小鼠大脑的神经元活动。结果突出了我们的方法在小鼠大脑浅层和深层同时进行动态成像的巨大潜力。
