Low‐photobleaching line‐scanning confocal microscopy using dual inclined beams
Low‐photobleaching line‐scanning confocal microscopy using dual inclined beams
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DOI：
10.1002/jbio.201900075
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发表时间：
2019-06
影响因子：
2.8
通讯作者：
Jialei Tang;K. Y. Han
中科院分区：
文献类型：
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作者：
Jialei Tang;K. Y. Han
共聚焦显微镜由于其高分辨率和光学切片能力而成为生物成像不可或缺的工具。然而，其缓慢的成像速度和严重的光漂白在很大程度上阻止了进一步的应用。在这里，我们提出了双倾斜光束线扫描（LS）共焦显微镜。与传统LS显微镜相比，我们成像方法的激发强度降低，使荧光观察时间延长2倍，同时保持良好的切片能力和单分子灵敏度。我们表征了我们的方法的性能，并将其应用于哺乳动物细胞的亚细胞成像和三维单分子RNA成像。
Confocal microscopy is an indispensable tool for biological imaging due to its high resolution and optical sectioning capability. However, its slow imaging speed and severe photobleaching have largely prevented further applications. Here, we present dual inclined beam line‐scanning (LS) confocal microscopy. The reduced excitation intensity of our imaging method enabled a 2‐fold longer observation time of fluorescence compared to traditional LS microscopy while maintaining a good sectioning capability and single‐molecule sensitivity. We characterized the performance of our method and applied it to subcellular imaging and three‐dimensional single‐molecule RNA imaging in mammalian cells.