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 as the ChipScope, we demonstrate successful culturing and imaging of fibroblasts, primary rat hippocampal neurons and axons of Xenopus retinal 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.

中文翻译：

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用于大视场神经元活细胞 TIRF 成像的波导成像平台。

通过波导实现的全内反射荧光显微镜 (TIRFM) 中的大视场 (FOV) 已被证明对于固定细胞上的单分子定位显微镜非常有益 [1,2]，并且也已被证明可用于短期实时成像强大的细胞类型[3-5]，但还没有针对脆弱的初级神经元，也没有在很长一段时间内。在这里，我们提出了一种基于波导的 TIRFM 设置，用于对要求严格的样品进行活细胞成像。使用开发的显微镜（称为ChipScope），我们展示了非洲爪蟾成纤维细胞、原代大鼠海马神经元和轴突的成功培养和成像视网膜神经节细胞（RGC）。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超大激发区域和卓越的照明均匀性，在神经科学应用中具有广泛的应用潜力。