Single-cell transfection by electroporation using an electrolyte/plasmid-filled capillary.

ABSTRACT: Single-cell transfection of adherent cells has been accomplished using single-cell electroporation (SCEP) with a pulled capillary. HEPES-buffered physiological saline solution containing pEGFP plasmid at a low concentration (0.16 approximately 0.78 microg/microL) filled a 15 cm long capillary with a tip opening of 2 microm. The electric field is applied to individual cells by bringing the tip close to the cell and subsequently applying one or two brief electric pulses. Many individual cells can thus be transfected with a small volume of plasmid-containing solution (approximately 1 microL). The extent of electroporation is determined by measuring the percentage loss of freely diffusing thiols (chiefly reduced glutathione) that have been derivatized with the fluorogenic ThioGlo 1. A mass tra

SUBMITTER: Wang M 

PROVIDER: S-EPMC2739742 | biostudies-literature | 2009 May

REPOSITORIES: biostudies-literature