Electroporationthe phenomenon and technical

  approach to experiments

Wroclaw, 24.4.2009

Peter KramarUniversity of LjubljanaFaculty of Electrical Engineeringhttp://lbk.fe.uni-lj.sipeter.kramar@fe.uni-lj.si

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!