超分辨率荧光成像对时间分辨率和长期稳定性的需求促使研究人员提高荧光探针的光稳定性。由于荧光团不可避免的光漂白,无论是引入过氧化物清除剂的自愈策略还是荧光团结构修饰抑制TICT形成的策略,都难以获得长期的超分辨率成像。缓冲荧光探针利用缓冲池中完整的探针不断替换目标中的光漂白探针,大大提高了光稳定性,实现了长时间稳定的动态超分辨率成像。但是缓冲能力是以减少靶标中荧光探针的数量为代价的,从而导致染色荧光强度低。在本文中,我们选择了具有高荧光亮度的脂滴探针BODIPY 493,来探索该探针在细胞中脂滴染色的动态过程。我们发现BODIPY 493由于脂滴中分子的聚集和高亮度,只需要非常低的激光功率进行脂滴成像,时空分辨率大大提高。更重要的是,我们发现BODIPY 493还具有一定的缓冲能力,这使得BODIPY 493能够用于脂滴动力学的超分辨率成像。这项工作提醒研究人员协调荧光探针的缓冲能力和亮度。我们发现BODIPY 493由于脂滴中分子的聚集和高亮度,只需要非常低的激光功率进行脂滴成像,时空分辨率大大提高。更重要的是,我们发现BODIPY 493还具有一定的缓冲能力,这使得BODIPY 493能够用于脂滴动力学的超分辨率成像。这项工作提醒研究人员协调荧光探针的缓冲能力和亮度。我们发现BODIPY 493由于脂滴中分子的聚集和高亮度,只需要非常低的激光功率进行脂滴成像,时空分辨率大大提高。更重要的是,我们发现BODIPY 493还具有一定的缓冲能力,这使得BODIPY 493能够用于脂滴动力学的超分辨率成像。这项工作提醒研究人员协调荧光探针的缓冲能力和亮度。这使得 BODIPY 493 能够用于脂滴动力学的超分辨率成像。这项工作提醒研究人员协调荧光探针的缓冲能力和亮度。这使得 BODIPY 493 能够用于脂滴动力学的超分辨率成像。这项工作提醒研究人员协调荧光探针的缓冲能力和亮度。
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BODIPY 493 acts as a bright buffering fluorogenic probe for super-resolution imaging of lipid droplet dynamics
The need for temporal resolution and long-term stability in super-resolution fluorescence imaging has motivated research to improve the photostability of fluorescent probes. Due to the inevitable photobleaching of fluorophores, it is difficult to obtain long-term super-resolution imaging regardless of the self-healing strategy of introducing peroxide scavengers or the strategy of fluorophore structure modification to suppress TICT formation. The buffered fluorogenic probe uses the intact probes in the buffer pool to continuously replace the photobleached ones in the target, which greatly improves the photostability and enables stable dynamic super-resolution imaging for a long time. But the buffering capacity comes at the expense of reducing the number of fluorescent probes in targets, resulting in low staining fluorescence intensity. In this paper, we selected BODIPY 493, a lipid droplet probe with high fluorescence brightness, to explore the dynamic process of lipid droplet staining of this probe in cells. We found that BODIPY 493 only needs very low laser power for lipid droplet imaging due to the high molecular accumulation in lipid droplets and the high brightness, and the spatiotemporal resolution is greatly improved. More importantly, we found that BODIPY 493 also has a certain buffering capacity, which enables BODIPY 493 to be used for super-resolution imaging of lipid droplet dynamics. This work reminds researchers to coordinate the buffering capacity and brightness of fluorogenic probes.