Adaptive-optics SLO imaging combined with widefield OCT and SLO enables precise 3D localization of fluorescent cells in the mouse retina.
Adaptive-optics SLO imaging combined with widefield OCT and SLO enables precise 3D localization of fluorescent cells in the mouse retina.
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DOI：
10.1364/boe.6.002191
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发表时间：
2015-06
影响因子：
3.4
通讯作者：
R. Zawadzki;Pengfei Zhang;A. Zam;Eric B. Miller;Mayank Goswami;Xinlei Wang;R. Jonnal;Sang-Hyuck Lee;D. Kim;J. Flannery;J. Werner;M. E. Burns;E. Pugh
中科院分区：
文献类型：
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作者：
R. Zawadzki;Pengfei Zhang;A. Zam;Eric B. Miller;Mayank Goswami;Xinlei Wang;R. Jonnal;Sang-Hyuck Lee;D. Kim;J. Flannery;J. Werner;M. E. Burns;E. Pugh
自适应光学扫描激光眼底镜(AO-SLO)最近被用来实现对小鼠视网膜的精细亚细胞分辨率成像。基于波前传感的声光通常将视野限制在几度视角内。因此，AO-SLO数据与更大范围的视网膜结构和细胞模式之间的关系可能很难评估。视网膜血管系统提供了一张大规模的3D地图，在活体成像期间可以在上面定位细胞和结构。位相方差OCT(PV-OCT)可以利用近红外光以无标记的方式高效地对血管系统进行成像，从而实现高精度的三维血管重建。我们将广域PV-OCT和SLO成像与AO-SLO反射和荧光成像相结合，在视网膜层内定位两种类型的荧光细胞：表达GFP的小胶质细胞，视网膜的驻留巨噬细胞，以及表达GFP的视锥感光细胞。我们详细描述了一种反射式无焦AO-SLO视网膜成像系统，该系统专为小鼠的高分辨率视网膜成像而设计。将该仪器的光学性能与其他最先进的基于AO的小鼠视网膜成像系统进行了比较。通过视网膜毛细血管造影术，包括血流速度分析，来表征新的AO器械的空间和时间分辨率。小胶质细胞和视锥感光细胞的深度分辨AO-SLO荧光图像与微血管和其他结构的469 nm和663 nm反射图像平行显示。文中还讨论了新仪器的其他应用。
Adaptive optics scanning laser ophthalmoscopy (AO-SLO) has recently been used to achieve exquisite subcellular resolution imaging of the mouse retina. Wavefront sensing-based AO typically restricts the field of view to a few degrees of visual angle. As a consequence the relationship between AO-SLO data and larger scale retinal structures and cellular patterns can be difficult to assess. The retinal vasculature affords a large-scale 3D map on which cells and structures can be located during in vivo imaging. Phase-variance OCT (pv-OCT) can efficiently image the vasculature with near-infrared light in a label-free manner, allowing 3D vascular reconstruction with high precision. We combined widefield pv-OCT and SLO imaging with AO-SLO reflection and fluorescence imaging to localize two types of fluorescent cells within the retinal layers: GFP-expressing microglia, the resident macrophages of the retina, and GFP-expressing cone photoreceptor cells. We describe in detail a reflective afocal AO-SLO retinal imaging system designed for high resolution retinal imaging in mice. The optical performance of this instrument is compared to other state-of-the-art AO-based mouse retinal imaging systems. The spatial and temporal resolution of the new AO instrumentation was characterized with angiography of retinal capillaries, including blood-flow velocity analysis. Depth-resolved AO-SLO fluorescent images of microglia and cone photoreceptors are visualized in parallel with 469 nm and 663 nm reflectance images of the microvasculature and other structures. Additional applications of the new instrumentation are discussed.