A Novel Electroporation System for Living Cell Staining and Membrane Dynamics Interrogation

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.

Dielectric Dispersion Modulated Sensing of Yeast Suspension Electroporation

“…The dielectric dispersion dynamics can be used as a design parameter for micro/nano dielectric dispersion modulated sensors. Those sensors may be used for industrial irreversible electroporation or lab on a chip electroporation device [9]. FEM simulation has been used to find the equivalent ε, which depends on the inhomogeneous media.…”

“…Techniques and sensing electroporation improve safety and optimization applications of PEFs. Recent progress on electroporation methods propose studies of PEF using nanoparticles [6], gels [7,8], microdevices [9], and electrospun PCL [10].…”

In Situ Deployment of Engineered Extracellular Vesicles into the Tumor Niche via Myeloid‐Derived Suppressor Cells

“…MDSC migration to the tumor microenvironment is mediated by an array of cytokines and chemokines, primarily CCL2, but also other CXC-motif chemokines. [63] While our data suggest that these recruitment mechanisms are likely maintained in engineered MDSCs, it is possible that the degree of engineered MDSC tropism toward diseased tissue versus off-target accumulation could be impacted by the method used to transfect the MDSCs (e.g., nonviral versus viral approaches), [64][65][66][67][68][69][70][71][72] and/or the type of cargo/transcript that is overexpressed in them. Thus, future studies should continue to evaluate how these parameters influence MDSC and MDSC-derived EV targeting to diseased tissues, as well as the potential consequences of off-target secretion of the EVs and corresponding therapeutic payloads in other tissues.…”

In situ electroporation of mammalian cells through SiO2 thin film capacitive microelectrodes

“…In alternative, there is a growing interest for methods of in-situ electroporation, which avoid the potentially detrimental enzymatic and mechanical treatments to detach cells growing in adhesion on a solid substrate. In particular, recent developments include miniaturized and on-chip integrated microsystems that selectively operate in-situ electroporation on subpopulations of cells in adhesion 7 – 9 . With respect to bulk electroporation, these devices enable to operate spatially confined transfections of preselected subpopulations of cells 10 , 11 or single cells 12 , 13 .…”