Abstract
A novel electroporation system was developed to introduce transient membrane pores to cells in a spatially and temporally controlled manner, allowing us to achieve fast electrotransfection and live cell staining as well as to systematically interrogate the dynamics of the cell membrane. Specifically, using this platform, we showed that both reversible and irreversible electroporation could be induced in the cell population, with nano-sized membrane pores in the former case being able to self-reseal in ~10 min. In addition, green fluorescent protein(GFP)-vinculin plasmid and 543 phalloidin have been delivered successively into fibroblast cells, which enables us to monitor the distinct roles of vinculin and F-actin in cell adhesion and migration as well as their possible interplay during these processes. Compared to conventional bulk electroporation and staining methods, the new system offers advantages such as low-voltage operation, cellular level manipulation and testing, fast and adjustable transfection/staining and real-time monitoring; the new system therefore could be useful in different biophysical studies in the future.
Highlights
Under a high-voltage electric field, transient pores can be created on the cell membrane which changes its conductivity and permeability, a process referred to as electroporation [1]
In addition to facilitating the formation of Focal adhesions (FAs), the forces generated by F-actin are responsible for driving other processes such as cell spreading and migration [19,20]
It has been found that once the magnitude or duration of the applied electric field is beyond a threshold level, damage on the cell membrane can be permanent, a phenomenon known as irreversible electroporation [28]
Summary
Under a high-voltage electric field, transient pores can be created on the cell membrane which changes its conductivity and permeability, a process referred to as electroporation [1]. Based on a micro-electroporation system, a recent study has shown that the electroporation efficiency of lung tumor cells is closely related to their resistance against commonly used anti-cancer drugs [6,7], indicating the possibility of utilizing electroporation in the classification and prognosis of cancer Another important application of electroporation is small particle delivery for drug loading [8], living cell staining or transfection [9], which is crucial for elucidating the biological roles/duties of different cellular components and organelles. Conventional methods of living cell staining rely on the proper characteristics (like reactivity and hydrophilicity) of staining molecules that allow them to pass through the membranes of living cells spontaneously [10] It usually takes hours for small dyes to enter the cell, making it challenging to achieve immediate and real-time staining and monitoring. The selective electroporation of bacteria was achieved by using metallodlectric Janus particles as electrodes [12]
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