Nano-localized single-cell nano-electroporation

• Published in: Lab on a chip Vol. 2; no. 22; pp. 4194 - 424
• Main Authors: Santra, Tuhin Subhra, Kar, Srabani, Chang, Hwan-You, Tseng, Fan-Gang
• Format: Journal Article
• Language: English
• Published: Cambridge Royal Society of Chemistry 01.01.2020
• Subjects: Biomolecules; Cell membranes; Cells (biology); Diagnostic systems; Electric fields; Electrodes; Electroporation; Field strength; Pressurization; Pulse duration
• ISSN: 1473-0197; 1473-0189
• DOI: 10.1039/d0lc00712a

The ability to deliver foreign cargos into single living cells is of great interest in cell biology and therapeutic research. Here, we have reported a single or multiple position based nano-localized single-cell nano-electroporation platform. The device consists of an array of triangular shape ITO nano-electrodes with a 70 nm gap between two nano-electrodes, each having a 40 nm tip diameter. The voltage is applied between nano-electrodes to generate an intense electric field, which electroporates multiple nano-localized regions of the targeted single-cell membrane, and biomolecules are gently delivered into cells by pressurizing pump flow, without affecting cell viability. The platform successfully delivers dyes, QDs, and plasmids into different cell types with the variation of field strength, pulse duration, and the number of pulses. This new approach allows us to analyze delivery of different biomolecules into single living cells with high transfection efficiency (>96%, for CL1-0 cells) and high cell viability (∼98%), which are potentially beneficial for cellular therapy and diagnostic purposes. We demonstrated nano-electroporation technique to create transient nano-holes at single or multiple nano-localized positions of a single-cell for a highly efficient intracellular delivery with high cell viability.