Measurement of molecular transport after electropermeabilization

Abstract

Measurements of molecular transport after electropermeabilization of biological membranes have been reported since the early days of electroporation research. Monitoring molecular transport not only can help reveal underlying mechanisms but also is critical for applications that aim to deliver molecules into cells via electric field exposures. However, questions remain open in part because of the size of the multi-dimensional experimental parameter spectrum that spans a wide range of electrical and biological factors. Quantitative measurements that address these questions are essential in guiding development of more accurate and predictive electropermeabilization models and in optimizing parameter sets for specific applications. This chapter introduces the main experimental considerations for quantitative measurements of molecular transport after electropermeabilization. A brief discussion of the basic theoretical equations governing molecular transport is followed by discussions of electrical exposure systems, measurement methods, calibration of fluorescence measurements to absolute quantities, and some examples. The chapter provides perspective on the wide range of experimental design decisions to be made and emphasizes the need for careful, quantitative, molecular transport measurements in order to test, improve, and guide models, experiments, and applications.