A waveguide imaging platform for live-cell TIRF imaging of neurons over large fields of view.
A waveguide imaging platform for live-cell TIRF imaging of neurons over large fields of view.
复制标题
用于大视场神经元活细胞 TIRF 成像的波导成像平台。
DOI：
10.1002/jbio.201960222
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发表时间：
2020
影响因子：
2.8
通讯作者：
Opstad IS
中科院分区：
文献类型：
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作者：
Opstad IS
通过波导的全内反射荧光显微镜（TIRFM）中的大视场（fov）已被证明对固定细胞的单分子定位显微镜非常有益[1,2]，并且也被证明可用于稳健细胞类型的短期活成像[3‐5]，但尚未用于脆弱的初级神经元或长时间成像。在这里，我们提出了一种基于波导的TIRFM装置，用于要求高的样品的活细胞成像。使用开发的显微镜，被称为ChipScope，我们成功地培养和成像成纤维细胞，原代大鼠海马神经元和异视视网膜神经节细胞（RGCs）的轴突。TIRFM提供的高对比度和柔和的照明模式，加上光子波导提供的特别大的激发区域和优越的照明均匀性，在神经科学应用中具有广泛的应用潜力。
Large fields of view (FOVs) in total internal reflection fluorescence microscopy (TIRFM) via waveguides have been shown to be highly beneficial for single molecule localisation microscopy on fixed cells [1,2] and have also been demonstrated for short‐term live‐imaging of robust cell types [3‐5], but not yet for delicate primary neurons nor over extended periods of time. Here, we present a waveguide‐based TIRFM set‐up for live‐cell imaging of demanding samples. Using the developed microscope, referred to asthe ChipScope, we demonstrate successful culturing and imaging of fibroblasts, primary rat hippocampal neurons and axons ofXenopusretinal ganglion cells (RGCs). The high contrast and gentle illumination mode provided by TIRFM coupled with the exceptionally large excitation areas and superior illumination homogeneity offered by photonic waveguides have potential for a wide application span in neuroscience applications.