Combined Numerical and Experimental Investigation of Localized Electroporation-Based Cell Transfection and Sampling

Abstract

Localized electroporation has evolved as an effective technology for the delivery of foreign molecules into cells while preserving their viability. Consequently, this technique has potential applications in sampling the contents of live cells and the temporal assessment of cellular states at the single-cell level. Although there have been numerous experimental reports on localized electroporation-based delivery, a lack of a mechanistic understanding of the process hinders its implementation in sampling. In this work, we develop a multiphysics model that predicts the transport of molecules into and out of the cell during localized electroporation. Based on the model predictions, we optimize experimental parameters such as buffer conditions, electric field strength, cell confluency, and density of nanochannels in the substrate for successful delivery and sampling via localized electroporation. We also identify that cell membrane tension plays a crucial role in enhancing both the amount and the uniformity of molecular transport, particularly for macromolecules. We qualitatively validate the model predictions on a localized electroporation platform by delivering large molecules (bovine serum albumin and mCherry-encoding plasmid) and by sampling an exogeneous protein (tdTomato) in an engineered cell line.

Nanofountain Probe Electroporation Enables Versatile Single‐Cell Intracellular Delivery and Investigation of Postpulse Electropore Dynamics

“…The maximum transport of calcein molecules across the cell membrane occurs at intermediate values of the pulse parameters. A similar trend was also demonstrated previously, [22,24,57] where the maximum transfection efficiency of a plasmid occurred for an intermediate value of the pulse voltage amplitude in multiple cell types.…”

“…[19] Finally, bulk electroporation reduces cell viability due to the high voltages applied and provides low precision over dosage. [20] During the last decade, we [21][22][23][24] and others [25,26] have shown that disadvantages associated with bulk electroporation can be overcome by focusing an electric field on a small region of the plasma membrane. Localized electroporation-owing to the confined electric field-also benefits from strong electrokinetic effects that can confer improved delivery.…”

Recent Developments in Cell Permeable Deubiquitinating Enzyme Activity-Based Probes

“…While generally used to transfect exogenous DNA into cells, this technology may be useful for the intracellular delivery of large molecules, such as Ub-Dha (Shi et al, 2018). An advantage of this method, particularly in comparison to poreforming toxins, is that the pores formed are very small and transient, so cell viability and functionality is usually preserved (Mukherjee et al, 2018). Consistent with this, the authors report normal cell morphology post-electroporation.…”

Nanoneedle Platforms: The Many Ways to Pierce the Cell Membrane

“…Using electric pulses, cell membranes can be transiently perforated so that a large variety of materials can be intracellularly delivered. In recent years, electroporation coupled with micro/nanostructures, [ 151–154 ] called as micro/nanoscale electroporation (MEP/NEP), has significantly improved cell viability while ensuring efficient transfection. These micro/nanostructure‐mediated local electric fields can more gently penetrate the cell membrane, and the resulting cell damage can be almost negligible.…”