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Waveguide-PAINT offers an open platform for large field-of-view super-resolution imaging

机译:Waveguide-PAINT为大视场超分辨率成像提供开放平台

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摘要

Super-resolution microscopies based on the localization of single molecules have been widely adopted due to their demonstrated performance and their accessibility resulting from open software and simple hardware. The PAINT method for localization microscopy offers improved resolution over photoswitching methods, since it is less prone to sparse sampling of structures and provides higher localization precision. Here, we show that waveguides enable increased throughput and data quality for PAINT, by generating a highly uniform ~100 × 2000 µm2 area evanescent field for TIRF illumination. To achieve this, we designed and fabricated waveguides optimized for efficient light coupling and propagation, incorporating a carefully engineered input facet and taper. We also developed a stable, low-cost microscope and 3D-printable waveguide chip holder for easy alignment and imaging. We demonstrate the capabilities of our open platform by using DNA-PAINT to image multiple whole cells or hundreds of origami structures in a single field of view.
机译:基于单分子定位的超分辨率显微技术已被广泛采用,这是由于它们具有开放式软件和简单硬件所表现出的性能和可访问性。定位显微镜的PAINT方法比光开关方法具有更高的分辨率,因为它不易出现稀疏的结构采样,并且定位精度更高。在这里,我们证明波导可以通过为TIRF照明产生高度均匀的〜100×2000 µm 2 van逝场来提高PAINT的吞吐量和数据质量。为此,我们设计并制造了经过优化的波导,以实现有效的光耦合和传播,并结合了精心设计的输入面和锥度。我们还开发了稳定,低成本的显微镜和3D可打印波导芯片支架,可轻松对准和成像。通过使用DNA-PAINT在单个视场中成像多个完整细胞或数百个折纸结构,我们演示了开放平台的功能。

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