Chip-based wide field-of-view nanoscopy
Chip-based wide field-of-view nanoscopy
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DOI：
10.1038/nphoton.2017.55
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发表时间：
2017-05-01
期刊：
影响因子：
35
通讯作者：
Ahluwalia, Balpreet S.
中科院分区：
文献类型：
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作者：
Diekmann, Robin;Helle, Oystein I.;Ahluwalia, Balpreet S.
目前的光学纳米技术使用复杂的显微镜进行成像，并使用简单的载玻片来保持样品。在这里，我们展示了相反的情况：使用复杂但可批量生产的光学芯片，它承载样品并为照明源提供波导，以及标准的低成本显微镜，通过两种不同的方法获得超分辨率图像。由具有高折射率对比度的材料组成的波导提供用于单分子切换和荧光激发的强倏逝场，从而实现基于芯片的单分子定位显微镜。此外，多模干涉图案引起空间荧光强度变化，从而实现基于波动的超分辨率成像。由于基于芯片的纳米显微镜将照明和检测光路分开，全内反射荧光激发在大视场内是可能的，最大可达0.5 mm x 0.5 mm。使用基于多色芯片的纳米显微镜，我们可视化肝窦内皮细胞中的开窗。
Present optical nanoscopy techniques use a complex microscope for imaging and a simple glass slide to hold the sample. Here, we demonstrate the inverse: the use of a complex, but mass-producible optical chip, which hosts the sample and provides a waveguide for the illumination source, and a standard low-cost microscope to acquire super-resolved images via two different approaches. Waveguides composed of a material with high refractive-index contrast provide a strong evanescent field that is used for single-molecule switching and fluorescence excitation, thus enabling chip-based single-molecule localization microscopy. Additionally, multimode interference patterns induce spatial fluorescence intensity variations that enable fluctuation-based super-resolution imaging. As chip-based nanoscopy separates the illumination and detection light paths, total-internal-reflection fluorescence excitation is possible over a large field of view, with up to 0.5 mm x 0.5 mm being demonstrated. Using multicolour chip-based nanoscopy, we visualize fenestrations in liver sinusoidal endothelial cells.