Single-step electrical field strength screening to determine electroporation induced transmembrane transport parameters

Gadi Blumrosen, Alireza Abazari, Alexander Golberg, Martin L. Yarmush, Mehmet Toner

Research output: Contribution to journalArticlepeer-review

Abstract

The design of effective electroporation protocols for molecular delivery applications requires the determination of transport parameters including diffusion coefficient, membrane resealing, and critical electric field strength for electroporation. The use of existing technologies to determine these parameters is time-consuming and labor-intensive, and often results in large inconsistencies in parameter estimation due to variations in the protocols and setups. In this work, we suggest using a set of concentric electrodes to screen a full range of electric field strengths in a single test to determine the electroporation-induced transmembrane transport parameters. Using Calcein as a fluorescent probe, we developed analytical methodology to determine the transport parameters based on the electroporation-induced pattern of fluorescence loss from cells. A monolayer of normal human dermal fibroblast (NHDF) cells were pre-loaded with Calcein and electroporated with an applied voltage of 750 V with 10 and 50 square pulses with 50 μs duration. Using our analytical model, the critical electric field strength for electroporation was found for the 10 and 50 pulses experiments. An inverse correlation between the field strength and the molecular transport time decay constant, and a direct correlation between field strength and the membrane permeability were observed. The results of this work can simplify the development of electroporation-assisted technologies for research and therapies.

Original languageEnglish
Pages (from-to)2041-2049
Number of pages9
JournalBiochimica et Biophysica Acta - Biomembranes
Volume1858
Issue number9
DOIs
StatePublished - 1 Sep 2016

Keywords

  • Cell membrane permeability
  • Concentric electrode system
  • Electroporation
  • High-throughput experiments
  • Transport parameters

Fingerprint

Dive into the research topics of 'Single-step electrical field strength screening to determine electroporation induced transmembrane transport parameters'. Together they form a unique fingerprint.

Cite this