Ultralong-Time Recovery and Low-Voltage Electroporation for Biological Cell Monitoring Enabled by a Microsized Multipulse Framework

Abstract

Long-term nondestructive monitoring of cells is of significant importance for understanding cell proliferation, cell signaling, cell death, and other processes. However, traditional monitoring methods are limited to a certain range of testing conditions and may reduce cell viability. Here, we present a microgap, multishot electroporation (M2E) system for monitoring cell recovery for up to ∼2 h using ∼5 V pulses and with excellent cell viability using a medium cell population. Electric field simulations reveal the bias-voltage- and gap-size-dependent electric field intensities in the M2E system. In addition to excellent transparency with low cell toxicity, the M2E system does not require specialized components, expensive materials, complicated fabrication processes, or cell manipulations; it just consists of a micrometer-sized pattern and a low-voltage square-wave generator. Ultimately, the M2E system can offer a long-term and nontoxic method of cell monitoring.

Transepithelial Electrical Impedance Increase Following Porous Substrate Electroporation Enables Label-Free Delivery

“…Perhaps the simplest explanation of the discrepancy is that there are two distinct mechanisms causing the impedance increase (Figure 5). The gradual increase we are measuring may be recovery and resealing of the cell membrane as reported in other studies ( 25, 29, 31, 32 ), but the response is shifted upward by a second mechanism that occurs rapidly. The second mechanism may be occurring only during the application of the electric field, which is a relatively brief period (∼0.2%) of the total observation period.…”

Ultralong recovery time in nanosecond electroporation systems enabled by orientational-disordering processes

“…This behavior is consistent with that of previous calculations of lipid-bilayer models with constant electric fields. 6,64,65 We found that by comparison with the model after injecting the first electric-field pulse, the model after applying the second electric-field pulse exhibits a larger degree of membrane permeabilization, as indicated by a larger pore population (Fig. 1e and f).…”

“…Emerging and critical applications in fields, including genetic engineering, molecular medicine and gene therapy, require high-performance electroporation techniques for operations, such as food sterilization, 1 genetic payload transfer, 2 tumor ablation 3,4 and cell monitoring. 5,6 Next-generation electroporation techniques have therefore been an important subject of research. Moreover, some of the electroporation devices have made inroads into Covid-19 vaccine technologies.…”

The Delivery of mRNA Vaccines for Therapeutics

“…Electroporation disrupts the cell membrane by generating electric shock for intracellular nucleic acid delivery. Delivery efficiency can be improved by adjusting voltage, capacitance, resistance and other factors, such as cell number, density, RNA quantity and pulse time [ 53 , 72 , 73 ]. Electroporation has been used for DC-based mRNA vaccines in clinical trials [ 74 , 75 , 76 ].…”