SERS study of single-live-cell electrical permeabilization dynamics via plasmonic nanotubes

Abstract

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.