基于等离子体纳米管的单个活细胞电透化动力学的SERS研究
SERS study of single-live-cell electrical permeabilization dynamics via plasmonic nanotubes
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中文总结 AI 辅助
本研究构建了带SERS功能的等离子体纳米管平台，监测单个活细胞电透化动力学，证实其可诱导质膜局部透化并追踪膜恢复，为单细胞分析等提供新方法。
中文摘要 AI 辅助
目前对微创方法分析单个活细胞内的过程和信号活动（包括致瘤细胞亚群的鉴定）的需求日益增长。然而，大多数传统分析方法需要裂解细胞，无法对同一细胞进行随时间的重复测量，或依赖可能扰乱细胞功能的外源标记和报告分子。已开发出多种基于垂直纳米管的应用，可通过低电压电穿孔实现活细胞监测与分析，但膜通透性的程度、持续时间及膜修复的动力学仍不明确。在此，我们构建了具备表面增强拉曼光谱（SERS）功能的等离子体平台，用于监测附着在直径100纳米、高度2微米的纳米管上的单个活细胞的电穿孔诱导膜通透性动力学。我们采用纤连蛋白作为细胞外基质（ECM）涂层，以促进细胞附着在纳米管上。使用荧光染料递送作为独立验证方法，我们证实所制备的纳米结构可诱导质膜的局部电透化，并能监测其后续恢复。我们进一步利用SERS追踪透化和膜封合过程中膜处的分子变化。SERS光谱为电穿孔及后续膜恢复过程中膜相关成分和ECM的变化提供了分子层面的见解。脉冲诱导分子变化的实时分析在单细胞分析、细胞内信号传导、细胞状态及细胞异质性（包括致瘤细胞亚群的鉴定）方面具有巨大潜力，该能力可助力新型生物传感检测的开发。
英文摘要
There is a growing demand for minimally invasive methods to analyze intracellular processes and signaling activities in individual living cells, including the identification of tumorigenic cell subpopulations. However, most conventional analytical methods require cell lysis, precluding repeated measurements in the same cell over time, or rely on exogenous labels and reporters that may perturb cellular function. Various applications based on vertical nanotubes have been developed that enable live cell monitoring and analysis by electroporation with low voltages. However, the extent and duration of membrane permeability and kinetics of membrane repair remain elusive. Here, we built a plasmonic platform with the capacity of surface enhanced Raman spectroscopy (SERS) to monitor the electroporation-induced membrane permeability dynamics in individual live cells attached onto 100-nm diameter nanotubes of 2 um height. Fibronectin was employed as extracellular matrix (ECM)-coating to facilitate cell attachment onto nanotubes. Using fluorescent-dye delivery as an independent validation method, we show that the fabricated nanostructures induce localized electrical permeabilization of the plasma membrane and enable monitoring of its subsequent recovery. We further use SERS to track molecular changes at the membrane during permeabilization and resealing. The SERS spectra provide molecular-level insight into changes in membrane-associated components and the ECM during electroporation and subsequent membrane recovery. Real time monitoring of pulse induced molecular changes holds great promise for characterizing intracellular signaling, cellular states, and cellular heterogeneity at the single cell level, including the identification of tumorigenic subpopulations. This capacity could facilitate the development of novel biosensing assay.
