Nondestructive evaluation of progressive neuronal changes in organotypic rat hippocampal slice cultures using ultrahigh-resolution optical coherence microscopy.
Nondestructive evaluation of progressive neuronal changes in organotypic rat hippocampal slice cultures using ultrahigh-resolution optical coherence microscopy.
复制标题
DOI：
10.1117/1.nph.1.2.025002
复制
发表时间：
2014-10
期刊：
影响因子：
5.3
通讯作者：
Zhou C
中科院分区：
文献类型：
--
作者：
Li F;Song Y;Dryer A;Cogguillo W;Berdichevsky Y;Zhou C
三维组织培养已被用作研究包括癫痫在内的各种疾病的有效模型。高通量、非破坏性技术对于快速评估与疾病相关的过程至关重要，例如进行性细胞死亡。本研究建立了一套纵向分辨率为1.5μm、横向分辨率为2.3μm的超高分辨率光学相干显微镜系统，用于评价器官型大鼠海马神经元的癫痫样损伤。通过将UHR-OCM图像与同一培养物的免疫共聚焦染色图像进行比较，证实了UHR-OCM显示神经组织中细胞的能力。分别于培养7、14、21、28天(DIVS)采集UHR-OCM图像，观察神经元损伤的进展。与DIV 7相比，在随后的时间点上，UHR-OCM图像的3D细胞计数和培养厚度在统计学上显著减少。在用犬尿酸(KYNA)处理的培养物中，与对照标本相比，细胞计数和培养物厚度的减少明显减少。这些结果表明，UHR-OCM能够对器官型海马神经元培养中的神经元死亡进行快速、无标记和非破坏性的评估。UHR-OCM与三维组织培养相结合，可能被证明是一种有希望的高通量筛选针对各种疾病的药物的工具。
Three-dimensional tissue cultures have been used as effective models for studying different diseases including epilepsy. High-throughput, non-destructive techniques are essential for rapid assessment of disease-related processes such as progressive cell death. In this study, an ultrahigh-resolution optical coherence microscopy (UHR-OCM) system with ~1.5-μm axial resolution and ~2.3-μm transverse resolution was developed to evaluate seizure-induced neuronal injury in organotypic rat hippocampal cultures. The capability of UHR-OCM to visualize cells in neural tissue was confirmed by comparison of UHR-OCM images with confocal immunostained images of the same cultures. In order to evaluate the progression of neuronal injury, UHR-OCM images were obtained from cultures on 7, 14, 21 and 28 days-in-vitro (DIVs). In comparison to DIV 7, statistically significant reductions in 3D cell count and culture thickness from UHR-OCM images were observed on subsequent time points. In cultures treated with kynurenic acid (KYNA), significantly less reduction in cell count and culture thickness was observed compared to control specimens. These results demonstrate the capability of UHR-OCM to perform rapid, label-free, and non-destructive evaluation of neuronal death in organotypic hippocampal cultures. UHR-OCM, in combination with three-dimensional tissue cultures, can potentially prove to be a promising tool for high throughput screening of drugs targeting various disorders.