electroporation the phenomenon and technical approach to experiments wroclaw, 24.4.2009 peter kramar...
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Electroporationthe phenomenon and technical
approach to experiments
Wroclaw, 24.4.2009
Peter KramarUniversity of LjubljanaFaculty of Electrical Engineeringhttp://[email protected]
Laboratory of Biocybernetics
http://lbk.fe.uni-lj.si
The phenomena
The phenomena
Applications
Reversibleelectroporation:
Cell fusion
Gene transfection
Drug delivery
Cancer treatment
Ireversible electroporation:
Food pasterizationTissue 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
Current clamp
Kalinowski et al, Meas Sci Technol 1995
Voltage clamp Kramar et al,
Bioelectrochemistry 2007
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
Montal M., Mueller P. PNAS 69: 3561-3566, 1972
Lipids
Aqueoussolution
Rising the level of aqueous solution
Discharge method Benz et al, ????
Capacitance measurement
Capacitance to period converter
Kalinowski et al, 1995
Voltage clamp - linear rising signal
Kramar et al., 2007
kU
Current clamp - linear rising signal
kI (A/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 TreatmentsInternational SCIENTIFIC WORKSHOP and POSTGRADUATE COURSE
http://www.cliniporator.com/ect
Thank you for your attention!