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Extended field-of-view single-molecule imaging by highly inclined swept illumination
Optica ( IF 8.4 ) Pub Date : 2018-08-28 , DOI: 10.1364/optica.5.001063
Jialei Tang , Kyu Young Han

Single-molecule imaging inside cells is a valuable tool to study subcellular structures, gene expression, and the dynamics of biomolecules. Here, we present highly inclined swept tile illumination microscopy. By sweeping a thin highly inclined and laminated optical sheet (HILO) with confocal slit detection, our method provides a twofold thinner illumination and greater than fortyfold larger imaging area than conventional HILO microscopy, enabling 3D single-molecule imaging with a high signal-to-background ratio. We demonstrate single-molecule mRNA imaging with a few probes or a single probe in cultured cells and mouse brain tissues, and video-rate live-cell imaging.

中文翻译:

通过高度倾斜的扫掠照明扩展视野的单分子成像

细胞内部的单分子成像是研究亚细胞结构,基因表达和生物分子动力学的有价值的工具。在这里,我们提出了高度倾斜的扫掠瓷砖照明显微镜。通过共焦缝检测扫掠高度倾斜且叠层的薄光学片(HILO),我们的方法提供的照明比传统HILO显微镜薄两倍,成像面积大四十倍,从而实现3D单分子成像并具有高信噪比背景比率。我们演示了几个分子或培养细胞和小鼠脑组织中的单个探针的单分子mRNA成像,以及视频速率活细胞成像。
更新日期:2018-09-21
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