Electroporation loading of membrane-impermeable molecules to investigate intra- and intercellular Ca2+ signaling.
Electroporation is a technique that temporarily induces pores in the plasma membranes of cells, thereby allowing plasma membrane-impermeable substances to enter the cells. This loading method requires an electrical drive circuit providing an electroporation signal, an electrode to apply the signal to a localized zone in a cell monolayer, and a special solution that has a low electrical conductivity. To avoid impairment of cell function and cell death from the electroporation procedure itself, the applied electrical signal should ideally be a high-frequency oscillating signal (50 kHz) without any direct current (DC) component. Here, we describe the detailed procedure of electroporation loading.
Department of Basic Medical Sciences, Physiology Group, Ghent University, 9000 Ghent, Belgium.
Elke
E
Decrock
Department of Basic Medical Sciences, Physiology Group, Ghent University, 9000 Ghent, Belgium.
Marijke
M
De Bock
Department of Basic Medical Sciences, Physiology Group, Ghent University, 9000 Ghent, Belgium.
Nan
N
Wang
Department of Basic Medical Sciences, Physiology Group, Ghent University, 9000 Ghent, Belgium.
Mélissa
M
Bol
Department of Basic Medical Sciences, Physiology Group, Ghent University, 9000 Ghent, Belgium.
Ashish K
AK
Gadicherla
Department of Basic Medical Sciences, Physiology Group, Ghent University, 9000 Ghent, Belgium.
Luc
L
Leybaert
D000818:Animals; D020013:Calcium Signaling; D002460:Cell Line; D002468:Cell Physiological Phenomena; D018274:Electroporation / Q000379:methods*; D006801:Humans