Abstract：
Long-term super-resolution imaging appears to be increasingly important for unraveling organelle dynamics at the nanoscale, but is challenging due to the need for highly photostable and environment-sensitive fluorescent probes. Here, we report a self-blinking fluorophore that achieved 12 nm spatial resolution and 20 ms time resolution under acidic lysosomal conditions. This fluorophore was successfully applied in super-resolution imaging of lysosomal dynamics over 40 min. The pH dependence of the dye during blinking made the fluorophore sensitive to lysosomal pH. This probe enables simultaneous dynamic and pH recognition of all lysosomes in the entire cell at the single-lysosome-resolved level, which allowed us to resolve whole-cell lysosome subpopulations based on lysosomal distribution, size, and luminal pH. We also observed a variety of lysosome movement trajectories and different types of interactions modes between lysosomes.
摘要：
长期的超分辨率成像对于解开纳米级的细胞器动力学似乎越来越重要，但由于需要高度光稳定和环境敏感的荧光探针而具有挑战性。这里,我们报告了一种自闪烁荧光团，在酸性溶酶体条件下达到12nm的空间分辨率和20ms的时间分辨率。该荧光团成功地应用于超过40分钟的溶酶体动力学的超分辨率成像。闪烁过程中染料的pH依赖性使荧光团对溶酶体pH敏感。该探针能够在单溶酶体分辨水平下同时动态和pH识别整个细胞中的所有溶酶体，这使我们能够根据溶酶体分布来解析全细胞溶酶体亚群，尺寸,和管腔pH值。我们还观察到各种溶酶体运动轨迹和溶酶体之间不同类型的相互作用模式。
