Photoactivatable Fluorophores for Bioimaging Applications
Photoactivatable Fluorophores for Bioimaging Applications
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用于生物成像应用的光活化荧光团
DOI：
10.1021/acsaom.3c00025
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发表时间：
2023
期刊：
影响因子：
--
通讯作者：
Raymo, Françisco M.
中科院分区：
文献类型：
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作者：
Zhang, Yang;Zheng, Yeting;Tomassini, Andrea;Singh, Ambarish Kumar;Raymo, Françisco M.
可光激活的荧光团提供了以精确的时间间隔仅在感兴趣的样本内的选定区域开启荧光的机会。这种时空荧光控制水平使成像方案的实施能够实时监视动态事件并以纳米分辨率可视化结构特征。这些变革性的成像方法正在为不同的细胞过程提供基本的见解，在生物学和医学上具有深远的意义。然而，目前的可光激活荧光团只有在激活事件之后才会发光，从而阻止了荧光图像的获取，从而阻止了样品在激活前的可视化。我们开发了一系列可光激活的荧光团，能够在发射态和光谱分辨荧光之间相互转换，而不是从非发射态切换到发射态。我们证明，我们的化合物可以实时监测在发育中的胚胎的细胞胚层中扩散的分子，以及沿着活线虫的肠道转移的聚合物珠。此外，它们还允许跟踪活细胞溶酶体内的单分子，并以纳米分辨率可视化这些细胞器。事实上，我们的可光激活的荧光团可能会演变成宝贵的分析工具，用于在分子水平上研究调节细胞功能和结构的基本因素。
Photoactivatable fluorophores provide the opportunity to switch fluorescence on exclusively in a selected area within a sample of interest at a precise interval of time. Such a level of spatiotemporal fluorescence control enables the implementation of imaging schemes to monitor dynamic events in real time and visualize structural features with nanometer resolution. These transformative imaging methods are contributing fundamental insights on diverse cellular processes with profound implications in biology and medicine. Current photoactivatable fluorophores, however, become emissive only after the activation event, preventing the acquisition of fluorescence images and, hence, the visualization of the sample prior to activation. We developed a family of photoactivatable fluorophores capable of interconverting between emissive states with spectrally resolved fluorescence, instead of switching from a nonemissive state to an emissive one. We demonstrated that our compounds allow the real-time monitoring of molecules diffusing across the cellular blastoderm of developing embryos as well as of polymer beads translocating along the intestinal tract of live nematodes. Additionally, they also permit the tracking of single molecules in the lysosomal compartments of live cells and the visualization of these organelles with nanometer resolution. Indeed, our photoactivatable fluorophores may evolve into invaluable analytical tools for the investigation of the fundamental factors regulating the functions and structures of cells at the molecular level.