Transcript Document
Electroporation the phenomenon and technical approach to experiments Peter Kramar University of Ljubljana Faculty of Electrical Engineering http://lbk.fe.uni-lj.si [email protected] Wroclaw, 24.4.2009 Laboratory of Biocybernetics http://lbk.fe.uni-lj.si The phenomena The phenomena Applications Reversible electroporation: Ireversible electroporation: Cell fusion Gene transfection Drug delivery Cancer treatment Food pasterization Tissue ablation Key issue The phenomena is already used (even in clinical applications) and it works well in several applications, but the basic principal are not clarified yet. Levels of electroporation • • • • • • Planar lipid bilayers Lipid vesicles Cells in suspension Attached cells Cell organelles Tissue Levels of electroporation • • • • • • Planar lipid bilayers Lipid vesicles Cells in suspension Attached cells Cell organelles Tissue Previous experiments Experimental systems Voltage clamp Kramar et al, Bioelectrochemistry 2007 Current clamp Kalinowski et al, Meas Sci Technol 1995 Measurement protocol Membrane formation • 1-palmitoyl 2-oleoyl phosphatidylcholine POPC • 1-palmitoyl 2-oleoyl phosphatidylserine POPS • POPC:POPS mixtures 1:3, 1:1and 3:1 Lipids Aqueous solution Rising the level of aqueous solution Montal M., Mueller P. PNAS 69: 3561-3566, 1972 Capacitance measurement Discharge method Benz et al, ???? Capacitance to period converter Kalinowski et al, 1995 Voltage clamp - linear rising signal kU Kramar et al., 2007 Current clamp - linear rising signal kI (mA/s) 0.03* 0.05* 0.1 0.2 0.5 1 4 8 10 20 November 15-22, 2009 Ljubljana, Slovenia Electroporation based Technologies and Treatments International SCIENTIFIC WORKSHOP and POSTGRADUATE COURSE http://www.cliniporator.com/ect Thank you for your attention!