Electroporation: High Frequency of Occurrence of a Transient High-Permeability State in Erythrocytes and Intact Yeast.

摘要

We present the first determinations of population distributions of macromolecule uptake due to electroporation, the percentage of cells which participate and, for the yeast, the subpopulation of cells whose membranes exhibit significant recovery following macromolecule uptake. Flow cytometry is used to measure the uptake of a first test molecule (green fluorescence, FITC-dextran; 70 kDa), and also, for the yeast, the subsequent uptake of a second, much smaller, test molecule (red fluorescence, propidium iodide; 660 Da), which provides a measure of membrane recovery. Electroporation is emerging as a general method for introduction of molecules into cells. Although microinjection is broadly applicable to mammalian cells and to protoplasts it is tedious, and generally inapplicable to intact microorganisms with cell walls. In contrast, a large number of cells can be simultaneously exposed to strong electric fields, which under proper conditions leads to electroporation, the occurrence of pores or membrane openings through which both ions and molecules can pass. Medicine, Reprints. (jes)