Electroporation-Assisted Surface-Enhanced Raman Detection for Long-Term, Label-Free, and Noninvasive Molecular Profiling of Live Single Cells

Molecule characterization of live single cells is greatly important in disease diagnoses and personalized treatments. Conventional molecule detection methods, such as mass spectrography, gene sequencing, or immunofluorescence, are usually destructive or labeled and unable to monitor the dynamic change of live cellular molecules. Herein, we propose an electroporation-assisted surface-enhanced Raman scattering (EP-SERS) method using a microchip to implement label-free, noninvasive, and continuous detections of the molecules of live single cells. The microchip containing microelectrodes with nanostructured EP-SERS probes has a multifunction of cell positioning, electroporation, and SERS detection. The EP-SERS method capably detects both the intracellular and extracellular molecules of live single cells without losing cell viability so as to enable long-term monitoring of the molecular pathological process in situ. We detect the molecules of single cells for two breast cancer cell lines with different malignancies (MCF-7 and MDA-MB-231), one liver cancer cell line (Huh-7), and one normal cell line (293T) using the EP-SERS method and classify these cell types to achieve high accuracies of 91.4-98.3% using their SERS spectra. Furthermore, 24 h continuous monitoring of the heterogeneous molecular responses of different cancer cell lines under doxorubicin treatment is successfully implemented using the EP-SERS method. This work provides a long-term, label-free, and biocompatible approach to simultaneously detect intracellular and extracellular molecules of live single cells on a chip, which would facilitate research and applications of cancer diagnoses and personalized treatments.