Electroporation serves as a powerful technique to introduce the exogenous nucleotides, DNA, RNA, proteins, dyes, and virus particles into cells. Through the effect of high intensity of electric field, the permeability of the cell membrane is instantaneously improved to absorb the exogenous molecules in surrounding medium. To protect the cell viability, ultralow-voltage electroporation techniques are well developed by versatile devices, and delivery efficiency is commonly assessed by label-based analysis by microscope-ImageJ or flow cytometry. However, accuracy and complexity of these analytical strategies still hinder efficient and precise biomedical studies in situ. Here we developed an intracellular delivery biosensing system by nanostrawed electroporation array (NEA) that can efficiently assess the universal electroporation performance by cell viability, delivery efficiency, and cell mortality. The intracellular delivery biosensing system consists of NEA, fluorescent microscope, and automated analysis software. Intracellular delivery biosensing system of electroporation quality is based on the enhanced fluorescent watershed segmentation (enhanced FWS) algorithm, which possessed low deviation (∼5%) and significantly shortened the operation time (∼8 s/10 images) in contrast to high deviation (∼13%) and long operation time (∼10 min/10 images) of conventional inaccurate, labor-intensive and time-consuming methods. It is believed that the intracellular delivery biosensing system will be a promising universal platform to assess nanodevice electroporation in the fields of cell biology, biotechnology, and medicine.

电穿孔是一种将外源性核苷酸、DNA、RNA、蛋白质、染料和病毒颗粒引入细胞的强大技术。通过高强度电场的作用，瞬间提高细胞膜的通透性，吸收周围介质中的外源性分子。为了保护细胞活力，超低电压电穿孔技术由多功能设备很好地开发，并且递送效率通常通过显微镜-ImageJ 或流式细胞术的基于标记的分析来评估。然而，这些分析策略的准确性和复杂性仍然阻碍了有效和精确的原位生物医学研究。在这里，我们通过纳米管电穿孔阵列 (NEA) 开发了一种细胞内递送生物传感系统，该系统可以通过细胞活力、递送效率和细胞死亡率有效评估通用电穿孔性能。细胞内递送生物传感系统由NEA、荧光显微镜和自动分析软件组成。电穿孔质量的细胞内递送生物传感系统基于增强型荧光分水岭分割（增强型 FWS）算法，与高偏差相比，该算法具有低偏差（~5%）并显着缩短了操作时间（~8 s/10 图像）。 〜13％）和传统的不准确，劳动密集型和耗时的方法的长操作时间（〜10分钟/ 10张图像）。