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Electroporationthe phenomenon and technical
approach to experiments
Wroclaw, 24.4.2009
Peter KramarUniversity of LjubljanaFaculty of Electrical Engineeringhttp://[email protected]
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Laboratory of Biocybernetics
http://lbk.fe.uni-lj.si
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The phenomena
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The phenomena
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Applications
Reversibleelectroporation:
Cell fusion
Gene transfection
Drug delivery
Cancer treatment
Ireversible electroporation:
Food pasterizationTissue ablation
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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.
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Levels of electroporation
• Planar lipid bilayers
• Lipid vesicles
• Cells in suspension
• Attached cells
• Cell organelles
• Tissue
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Levels of electroporation
• Planar lipid bilayers
• Lipid vesicles
• Cells in suspension
• Attached cells
• Cell organelles
• Tissue
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Previous experiments
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Experimental systems
Current clamp
Kalinowski et al, Meas Sci Technol 1995
Voltage clamp Kramar et al,
Bioelectrochemistry 2007
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Measurement protocol
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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
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Discharge method Benz et al, ????
Capacitance measurement
Capacitance to period converter
Kalinowski et al, 1995
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Voltage clamp - linear rising signal
Kramar et al., 2007
kU
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Current clamp - linear rising signal
kI (A/s)
0.03*
0.05*
0.1
0.2
0.5
1
4
8
10
20
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November 15-22, 2009
Ljubljana, Slovenia
Electroporation based Technologies and TreatmentsInternational SCIENTIFIC WORKSHOP and POSTGRADUATE COURSE
http://www.cliniporator.com/ect
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Thank you for your attention!