Scanning electroporation of selected areas of adherent cell cultures.
We present a computer-controlled scanning electroporation method. Adherent cells are electroporated using an electrolyte-filled capillary in contact with an electrode. The capillary can be scanned over a cell culture and locally deliver both an electric field and an electroporation agent to the target area without affecting surrounding cells. The instantaneous size of the targeted area is determined by the dimensions of the capillary. The size and shape of the total electroporated area are defined by these dimensions in combination with the scanning pattern. For example, striped and serpentine patterns of electroporated cells in confluent cultures can be formed. As it is easy to switch between different electroporation agents, the method is suitable for design of cell cultures with complex composition. Finite element method simulations were used to study the spatial distributions of the electric field and the concentration of an electroporation agent, as well as the fluid dynamics related to scanning and flow of electroporation agent from the capillary. The method was validated for transfection by introduction of a 9-base-pair-long randomized oligonucleotide into PC12 cells and a pmaxGFP plasmid coding for green fluorescent protein into CHO and WSS cells.
Olofsson
Jessica
J
Department of Physical Chemistry, Chalmers University of Technology, Kemivägen 10, SE-412 96 Gothenburg, Sweden.
Levin
Mikael
M
Strömberg
Anette
A
Weber
Stephen G
SG
Ryttsén
Frida
F
Orwar
Owe
O
D000818:Animals; D016466:CHO Cells; D002448:Cell Adhesion / Q000502:physiology; D018929:Cell Culture Techniques / Q000379:methods*; D002460:Cell Line; D002468:Cell Physiological Phenomena; D006224:Cricetinae; D003412:Cricetulus; D004573:Electrolytes / Q000737:chemistry; D018274:Electroporation / Q000379:methods*; D020342:Finite Element Analysis; D049452:Green Fluorescent Proteins / Q000235:genetics / Q000378:metabolism; D016716:PC12 Cells; D010957:Plasmids / Q000235:genetics; D051381:Rats; D015203:Reproducibility of Results; D014162:Transfection / Q000379:methods
D004573:Electrolytes; D049452:Green Fluorescent Proteins