Imaging and quantifying ganglion cells and other transparent neurons in the living human retina.
Imaging and quantifying ganglion cells and other transparent neurons in the living human retina.
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DOI：
10.1073/pnas.1711734114
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发表时间：
2017-11-28
影响因子：
11.1
通讯作者：
Miller DT
中科院分区：
文献类型：
--
作者：
Liu Z;Kurokawa K;Zhang F;Lee JJ;Miller DT
神经节细胞是视网膜神经回路的主要组成部分，但在活体人眼中难以观察和量化。在这里，我们展示了一种光学显微镜模式，不仅揭示了这些细胞的胞体，而且还揭示了它们的3D包装几何形状，主要亚型和向其他神经元的空间投影。该方法提供了组成视网膜的神经元、神经胶质和血管的丰富织锦的一瞥，从而暴露了视觉信息的神经处理的解剖学基底。在临床上，活体眼睛中视网膜神经元的高分辨率图像有望改善视网膜疾病中神经节细胞和其他神经元损失的诊断和评估治疗。神经节细胞（GCs）是视网膜神经回路的基础，处理光感受器信号以通过它们的轴突传输到大脑。然而，关于它们在视觉中的作用以及它们对导致失明的疾病的脆弱性，仍然有很多未知之处。一个主要的瓶颈是我们无法观察GC及其在活体人眼中的退化。尽管二十年来光学技术的发展，以成像细胞在活的人类视网膜，GC仍然难以捉摸，由于其高的光学透明度。传统的成像主要使用单散射光，以揭示GC失败，导致了对多重散射，荧光，双光子和相位成像技术，以提高GC的对比度的重点。在这里，我们表明，单散射光实际上携带大量的信息，揭示GC胞体，轴突和其他视网膜神经元，并允许其定量分析。我们对GC层胞体进行形态测量，包括GC投射到光感受器和识别主要GC亚型，甚至在神经纤维下方。我们通过以下方法获得了单散射图像：（i）将自适应光学与光学相干断层扫描结合以避免眼睛的光学模糊;（ii）进行3D亚细胞图像配准以避免运动模糊;以及（iii）使用胞体内的细胞器运动性作为内在对比剂。此外，离焦成像提供了将个体GC空间映射到底层无长突、双极、水平、感光器和视网膜色素上皮细胞的潜力，从而暴露了视觉信息的神经处理的解剖学基底。这种成像方式也是用于改善视网膜疾病的临床诊断和评估治疗的工具。
Ganglion cells are the primary building block of retinal neural circuitry, but have been elusive to observe and quantify in the living human eye. Here, we show a light microscopy modality that reveals not only the somas of these cells, but also their 3D packing geometry, primary subtypes, and spatial projection to other neurons. The method provides a glimpse of the rich tapestry of neurons, glia, and blood vessels that compose the retina, thus exposing the anatomical substrate for neural processing of visual information. Clinically, high-resolution images of retinal neurons in living eyes hold promise for improved diagnosis and assessing treatment of ganglion cell and other neuron loss in retinal disease. Ganglion cells (GCs) are fundamental to retinal neural circuitry, processing photoreceptor signals for transmission to the brain via their axons. However, much remains unknown about their role in vision and their vulnerability to disease leading to blindness. A major bottleneck has been our inability to observe GCs and their degeneration in the living human eye. Despite two decades of development of optical technologies to image cells in the living human retina, GCs remain elusive due to their high optical translucency. Failure of conventional imaging—using predominately singly scattered light—to reveal GCs has led to a focus on multiply-scattered, fluorescence, two-photon, and phase imaging techniques to enhance GC contrast. Here, we show that singly scattered light actually carries substantial information that reveals GC somas, axons, and other retinal neurons and permits their quantitative analysis. We perform morphometry on GC layer somas, including projection of GCs onto photoreceptors and identification of the primary GC subtypes, even beneath nerve fibers. We obtained singly scattered images by: (i) marrying adaptive optics to optical coherence tomography to avoid optical blurring of the eye; (ii) performing 3D subcellular image registration to avoid motion blur; and (iii) using organelle motility inside somas as an intrinsic contrast agent. Moreover, through-focus imaging offers the potential to spatially map individual GCs to underlying amacrine, bipolar, horizontal, photoreceptor, and retinal pigment epithelium cells, thus exposing the anatomical substrate for neural processing of visual information. This imaging modality is also a tool for improving clinical diagnosis and assessing treatment of retinal disease.
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DOI：
10.1002/cne.23995
发表时间：
2016-10-01
期刊：
The Journal of comparative neurology
影响因子：
--
作者：
Liao HW;Ren X;Peterson BB;Marshak DW;Yau KW;Gamlin PD;Dacey DM
通讯作者：
Dacey DM
影响因子：
3.5
作者：
Gualda, Emilio J.;Bueno, Juan M.;Artal, Pablo
通讯作者：
Artal, Pablo
影响因子：
11.2
作者：
CURCIO, CA;DRUCKER, DN
通讯作者：
DRUCKER, DN
影响因子：
5.7
作者：
Galetta, Kristin M.;Calabresi, Peter A.;Balcer, Laura J.
通讯作者：
Balcer, Laura J.
影响因子：
4.4
作者：
Liu Z;Kocaoglu OP;Miller DT
通讯作者：
Miller DT