Applications of electroporation of adherent cells in situ, on a partly conductive slide.
Nontraumatic, simple, and reproducible procedures for the introduction of nonpermeant molecules into adherent mammalian cells by in situ electroporation are described. Cells are grown on a glass slide, half of which is coated with electrically conductive, optically transparent, indium-tin oxide. An electric pulse is applied in the presence of the molecules to be introduced, and their effect on the cellular phenotype can be observed. The cells growing on the nonconductive side of the slide do not receive any pulse and serve as controls. Careful adjustment of electric field strength can achieve the introduction of the molecules into essentially 100% of the cells, and this treatment causes no detectable disruption to cellular metabolism. This is applied in the presence of the fluorescent dye, Lucifer yellow, causing its penetration into the cells growing on the conductive half of the slide. The migration of the dye to the nonelectroporated cells growing on the nonconductive area is microscopically observed under fluorescence illumination.
Raptis
L H
LH
Department of Microbiology and Immunology, Queen's University, Kingston, Ontario, Canada.
Brownell
H L
HL
Liu
S K
SK
Firth
K L
KL
MacKenzie
L W
LW
Stiles
C D
CD
Alberta
J A
JA
D000818:Animals; D001709:Biotechnology; D002448:Cell Adhesion; D002460:Cell Line; D004553:Electric Conductivity; D018274:Electroporation / Q000295:instrumentation / Q000379:methods*; D005069:Evaluation Studies as Topic; D005898:Glass; D007204:Indium; D007546:Isoquinolines; D051379:Mice; D051381:Rats; D013552:Swine; D017971:Tin Compounds
D007546:Isoquinolines; D017971:Tin Compounds; D007204:Indium; C017475:lucifer yellow; C045358:stannic oxide