PDF(Pubmed)
Abstract：
Controlling chemical processes in live cells is a challenging task. The spatial heterogeneity of biochemical reactions in cells is often overlooked by conventional means of incubating cells with desired chemicals. A comprehensive understanding of spatially diverse biochemical processes requires precise control over molecular activities at the subcellular level. Herein, a closed-loop optoelectronic control system is developed that allows the manipulation of biomolecular activities in live cells at high spatiotemporal precision. Chemical-selective fluorescence signals are utilized to command lasers that trigger specific chemical processes or control the activation of photoswitchable inhibitors at desired targets. This technology is fully compatible with laser scanning confocal fluorescence microscopes. The authors demonstrate selective interactions of a 405 nm laser with targeted organelles and simultaneous monitoring of cell responses by fluorescent protein signals. Notably, blue laser interaction with the endoplasmic reticulum leads to a more pronounced reduction in cytosolic green fluorescent protein signals in comparison to that with nuclei and lipid droplets. Moreover, when combined with a photoswitchable inhibitor, microtubule polymerization is selectively inhibited within the subcellular compartments. This technology enables subcellular spatiotemporal optical manipulation over chemical processes and drug activities, exclusively at desired targets, while minimizing undesired effects on non-targeted locations.
摘要：
控制活细胞中的化学过程是一项具有挑战性的任务。细胞中生化反应的空间异质性通常被用所需化学物质孵育细胞的常规手段所忽略。全面了解空间上不同的生化过程需要对亚细胞水平的分子活动进行精确控制。在这里，开发了一种闭环光电控制系统，可以以高时空精度操纵活细胞中的生物分子活动。利用化学选择性荧光信号来命令激光,所述激光触发特定化学过程或控制在所需靶标处的光可切换抑制剂的活化。该技术与激光扫描共聚焦荧光显微镜完全兼容。作者展示了405nm激光与靶向细胞器的选择性相互作用，并通过荧光蛋白信号同时监测细胞反应。值得注意的是,与细胞核和脂滴相比，蓝色激光与内质网的相互作用导致胞浆绿色荧光蛋白信号更明显的减少。此外，当与光可切换抑制剂结合时，c在亚细胞区室中被选择性抑制。这项技术能够对化学过程和药物活动进行亚细胞时空光学操纵，完全在预期目标上，同时最大限度地减少对非目标位置的不良影响。
