1. Electroporation the phenomenon and technical approach to experiments Wroclaw, 24.4.2009 Peter Kramar University of Ljubljana Faculty of Electrical Engineering http://lbk.fe.uni-lj.si peter.kramar@fe.uni-lj.si

2. Laboratory of Biocybernetics http://lbk.fe.uni-lj.si

3. The phenomena

5. Applications Reversible electroporation: Cell fusion Gene transfection Drug delivery Cancer treatment Ireversible electroporation: Food pasterization Tissue ablation

6. 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.

7. Levels of electroporation Planar lipid bilayers Lipid vesicles Cells in suspension Attached cells Cell organelles Tissue

8. Levels of electroporation Planar lipid bilayers Lipid vesicles Cells in suspension Attached cells Cell organelles Tissue

9. Previous experiments

10. Experimental systems Current clamp Kalinowski et al, Meas Sci Technol 1995 Voltage clamp Kramar et al, Bioelectrochemistry 2007

12. Measurement protocol

13. Membrane formation 1-palmitoyl 2-oleoyl phosphatidylcholine POPC 1-palmitoyl 2-oleoyl phosphatidylserine POPS POPC:POPS mixtures 1:3, 1:1and 3:1 Montal M., Mueller P. PNAS 69: 3561-3566, 1972 Lipids Aqueous solution Rising the level of aqueous solution

14. Discharge method Benz et al, ???? Capacitance measurement Capacitance to period converter Kalinowski et al, 1995

15. Voltage clamp - linear rising signal Kramar et al., 2007 k U

16. Current clamp - linear rising signal k I (  A/s) 0.03* 0.05* 0.10.1 0.20.2 0.50.5 1 4 8 10 20

18. November 15-22, 2009 Ljubljana, Slovenia Electroporation based Technologies and Treatments International SCIENTIFIC WORKSHOP and POSTGRADUATE COURSE http://www.cliniporator.com/ect

19. Thank you for your attention!