Light-sheet microscopy in the near-infrared II window
Light-sheet microscopy in the near-infrared II window
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DOI：
10.1038/s41592-019-0398-7
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发表时间：
2019-06-01
期刊：
影响因子：
48
通讯作者：
Dai, Hongjie
中科院分区：
文献类型：
--
作者：
Wang, Feifei;Wan, Hao;Dai, Hongjie
由于光散射的影响，对活体哺乳动物进行高时空分辨率的非侵入性深部组织三维光学成像具有挑战性。我们开发了近红外II（1，000 - 1，700 nm）光片显微镜，激发和发射分别高达约1，320 nm和1，700 nm，用于光学切片，穿透深度约为750 μ m，通过活组织，无需侵入性手术，在甘油清除的脑组织中深度约为2 mm。正常和倾斜配置的近红外II光片显微镜能够通过完整组织对活小鼠进行体内成像，揭示肿瘤微循环中的异常血流和T细胞运动，并以细胞分辨率绘制出肿瘤中的程序性死亡配体1和程序性细胞死亡蛋白1。通过完整小鼠头部的三维成像解析了颅骨和脑皮质之间的血管通道，并允许监测巨噬细胞和小胶质细胞向创伤性脑损伤部位的募集。
Non-invasive deep-tissue three-dimensional optical imaging of live mammals with high spatiotemporal resolution is challenging owing to light scattering. We developed near-infrared II (1,000-1,700 nm) light-sheet microscopy with excitation and emission of up to approximately 1,320 nm and 1,700 nm, respectively, for optical sectioning at a penetration depth of approximately 750 mu m through live tissues without invasive surgery and at a depth of approximately 2 mm in glycerol-cleared brain tissues. Near-infrared II light-sheet microscopy in normal and oblique configurations enabled in vivo imaging of live mice through intact tissue, revealing abnormal blood flow and T-cell motion in tumor microcirculation and mapping out programmed-death ligand 1 and programmed cell death protein 1 in tumors with cellular resolution. Three-dimensional imaging through the intact mouse head resolved vascular channels between the skull and brain cortex, and allowed monitoring of recruitment of macrophages and microglia to the traumatic brain injury site.