文献类型：期刊文献

英文题名：Specially Resolved Single Living Cell Perfusion and Targeted Fluorescence Labeling Based on Nanopipettes

作者：Lv, Jian[1];Wang, Xiao-Yuan[1];Zhou, Xin-Yue[1];Li, Da-Wei[1];Qian, Ruo-Can[1]

机构：[1]East China Univ Sci & Technol, Feringa Nobel Prize Scientist Joint Res Ctr, Frontiers Sci Ctr Materiobiol & Dynam Chem, Sch Chem & Mol Engn,Key Lab Adv Mat,Joint Int Lab, Shanghai 200237, Peoples R China

年份：2022

卷号：94

期号：40

起止页码：13860

外文期刊名：ANALYTICAL CHEMISTRY

收录：;EI(收录号：20224112848519);WOS:【SCI-EXPANDED(收录号:WOS:000862794000001)】；

基金：This research was supported by National Natural Science Foundation of China (21977031, 21777041), the China Postdoctoral Science Foundation (2021M701195), Shanghai Science and Technology Committee (19ZR1472300, 22ZR1416800), and the Fundamental Research Funds for the Central Universities. The authors thank Research Center of Analysis and Test of East China University of Science and Technology for the help on the characterization.

语种：英文

外文关键词：Brownian movement - Cell culture - Cell proliferation - Electroosmosis - Fluorescence - Gene transfer - Nanotechnology

摘要：Targeted delivery and labeling of single living cells in heterogeneous cell populations are of great importance to understand the molecular biology and physiological functions of individual cells. However, it remains challenging to perfuse fluorescence markers into single living cells with high spatial and temporal resolution without interfering neighboring cells. Here, we report a single cell perfusion and fluorescence labeling strategy based on nanoscale glass nanopipettes. With the nanoscale tip hole of 100 nm, the use of nanopipettes allows special perfusion and high-resolution fluorescence labeling of different subcellular regions in single cells of interest. The dynamic of various fluorescent probes has been studied to exemplify the feasibility of nanopipette-dependent targeted delivery. According to experimental results, the cytoplasm labeling of Sulfo-Cyanine5 and fluorescein isothiocyanate is mainly based on the Brownian movement due to the dyes themselves and does not have a targeting ability, while the nucleus labeling of 4 ',6-diamidino-2-phenylindole (DAPI) is originated from the adsorption between DAPI and DNA in the nucleus. From the finite element simulation, the precise manipulation of intracellular delivery is realized by controlling the electro-osmotic flow inside the nanopipettes, and the different delivery modes between nontargeting dyes and nucleus-targeting dyes were compared, showcasing the valuable ability of nanopipette-based method for the analysis of specially defined subcellular regions and the potential applications for single cell surgery, subcellular manipulation, and gene delivery.