中文 | English
祝贺钱若灿研究员等在《Anal. Chem.》发表研究论文

Ultra-Fast Mapping of Subcellular Domain via Nanopipette Based Electroosmotically Modulated Delivery into a Single Living Cell

基于超微玻璃纳米电极内电渗流调控的亚细胞域超快成像

Ruo-Can Qian#, Jian Lv#, and Yi-Tao Long*

钱若灿#, 吕键#, 龙亿涛*


Journal: Analytical Chemistry. 2018, 90, 13744−13750 DOI: 10.1021/acs.analchem.8b04159

摘要:近年来,单细胞检测方法发展迅速。然而,在单细胞水平上精准探测亚细胞域仍是一个挑战。本工作提出一种针对亚细胞域的快速成像方法,通过超微石英玻璃纳米电极(尖端直径~100 nm),可在20 s内将多种荧光分子精确注射到单个活细胞内。该方法可进一步用于监测经过二甲双胍处理的单个MCF-7乳腺癌细胞中线粒体膜电位的丧失过程。进一步,本工作利用理论计算和有限元模拟,对纳米管内的主要驱动力电渗流场进行了计算。计算结果表明,在工作范围内,纳米管尖端附近存在一个限域的浓度分布层。本方法实现了多种探针分子的无损、一步精准单细胞注射,具有良好的多通道单细胞成像效果,为理解单细胞到亚细胞域范围内的生化过程提供了有力工具。


Abstract: Recently, a variety of strategies have been developed for single-cell detection. However, the precise probing of the given area at single cell level is still a challenge. Here, we put forward a rapid and targeted imaging approach for the mapping of subcelluar domain, which realizes the precise injection of multi-fluorescence into a single living cell via an ultra-small quartz capillary nanopipette (~100 nm), and can successfully transport different fluorescent probe molecules to the pointing subcellullar area around the tip in the cytoplasm within 20 seconds. This method is also applied for monitoring the loss of intracellular mitochondrial membrane potential under the treatment of metformin in the single MCF-7 breast cancer cell. The major driven force in the nanopipette, electroosmotic flow, is evaluated by theory calculation method and finite element simulations, and the solution indicates a confined solute distribution profile around the tip within the working range. Overall, the nanopipette approach realizes the precise and simultaneous delivery of multiple probe molecules into the single living cell through the electroosmotically modulated, non-destructive, and one-step injection, which is especially powerful and convenient for the multi-channel single cell imaging and monitoring, indicating favorable potential for understanding, mornitoring, and controlling the biological processes from the single cell to subcellular organelles.

Figure 1. Schematic illustration of the electoosmotic delivery of multi-fluorescent molecules into a single living cell.

1. 基于超微玻璃纳米电极内电渗流调控的单细胞荧光灌注



原文链接:https://pubs.acs.org/doi/10.1021/acs.analchem.8b04159