Nano-Electro-Sampling and Intelligent Analysis System for Real-Time Gene Profiling of Living Cells.

Accurate intracellular genetic profiling of living cells enables dynamic monitoring of therapeutic response, facilitating timely treatment optimization. However, achieving real-time gene analysis while maintaining cell viability remains a persistent challenge in existing methodologies. In this work, a Nano-Electro-Sampling and Intelligent Analysis (Nesta) system is developed, which incorporates a portable biochip for living cell sampling and on-chip gene identification with an intelligent image analysis program for accurate gene quantification. The sampling processing utilizes nanopore-focused electric fields to safely perforate cell membrane and extract intracellular genes. Subsequent sample transfer is effectively realized by chip rotation, which minimizes sample loss and contamination. The detachable detection area satisfies both temperature requirements for gene amplification and cell viability maintenance for sequential sampling. Integrated with intelligent image analysis program, this system enables precise quantitative analysis of smartphone-captured images. Validated across 50 primary lung cancer cell samples, Nesta system demonstrates over 90% detection concordance with gene sequencing and establishes significant correlation between rebound expression of EGFR mutant genes and acquired drug resistance (AUC = 0.95). This system achieves real-time gene analysis of living cells, offering a transformative approach in therapeutic monitoring.