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Impedance Spectroscopy for Toxicity Testing Martin Dominguez Cronus Technologies Ltd.
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Impedance Spectroscopy for Toxicity Testing Martin Dominguez Cronus Technologies Ltd.

Feb 23, 2016

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Impedance Spectroscopy for Toxicity Testing Martin Dominguez Cronus Technologies Ltd. Barrier Forming Tissue. Epithelial and endothelial cell layers form selectively permeable barriers. The Concept. Growing cell layers on permeable membranes. The Concept. - PowerPoint PPT Presentation
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Page 1: Impedance Spectroscopy for Toxicity Testing Martin Dominguez Cronus  Technologies Ltd.

Impedance Spectroscopy for Toxicity Testing

Martin DominguezCronus Technologies Ltd.

Page 2: Impedance Spectroscopy for Toxicity Testing Martin Dominguez Cronus  Technologies Ltd.

Barrier Forming TissueEpithelial and endothelial cell layers form selectively permeable barriers

Page 3: Impedance Spectroscopy for Toxicity Testing Martin Dominguez Cronus  Technologies Ltd.

The ConceptGrowing cell layers on permeable membranes

Page 4: Impedance Spectroscopy for Toxicity Testing Martin Dominguez Cronus  Technologies Ltd.

The ConceptMeasuring the electrical impedance of epithelial and endothelial cell layers under physiological conditions

Page 5: Impedance Spectroscopy for Toxicity Testing Martin Dominguez Cronus  Technologies Ltd.

Understanding Impedance SpectraContributions stemming from CPE and RMED

Page 6: Impedance Spectroscopy for Toxicity Testing Martin Dominguez Cronus  Technologies Ltd.

Understanding Impedance SpectraThe formation of a plateau at mid-range frequencies is attributable to the cell layer’s resistance TER and capacitance CCL

Page 7: Impedance Spectroscopy for Toxicity Testing Martin Dominguez Cronus  Technologies Ltd.

Understanding Impedance SpectraAn increase in the resistance TER shifts the plateau upwards

Page 8: Impedance Spectroscopy for Toxicity Testing Martin Dominguez Cronus  Technologies Ltd.

Understanding Impedance SpectraAn increase in the capacitance CCL narrows the plateau

Page 9: Impedance Spectroscopy for Toxicity Testing Martin Dominguez Cronus  Technologies Ltd.

Time-Lapse Impedance Spectroscopy

Page 10: Impedance Spectroscopy for Toxicity Testing Martin Dominguez Cronus  Technologies Ltd.

The Setup

Page 11: Impedance Spectroscopy for Toxicity Testing Martin Dominguez Cronus  Technologies Ltd.

The Setup

Page 12: Impedance Spectroscopy for Toxicity Testing Martin Dominguez Cronus  Technologies Ltd.

cellZscopeAutomated cell monitoring device

Page 13: Impedance Spectroscopy for Toxicity Testing Martin Dominguez Cronus  Technologies Ltd.

Monitoring of MDCK-II CellsFormation of a differentiated cell monolayer with established cell-cell junctions

Page 14: Impedance Spectroscopy for Toxicity Testing Martin Dominguez Cronus  Technologies Ltd.

cellZscope vs. „Chopstick“ ElectrodesComparing TER values measured on MDCK-II cell layers

Page 15: Impedance Spectroscopy for Toxicity Testing Martin Dominguez Cronus  Technologies Ltd.

Long Term Monitoring of Caco-2 CellsComparing growth dynamics of different cell lines

Page 16: Impedance Spectroscopy for Toxicity Testing Martin Dominguez Cronus  Technologies Ltd.

0

20

40

60

80

100

120

140

0.0 0.5 1.0 1.5 2.0 2.5

TER

(%)

Ti me (h)

Control1mM EGTA2mM EGTA4mM EGTA

EGTA

re

move

d

EGTA

adde

d

EDTA Treatment of MDCK-II CellsEffects on the tight junction network

Page 17: Impedance Spectroscopy for Toxicity Testing Martin Dominguez Cronus  Technologies Ltd.

MBCD Treatment of MDCK-II CellsDose dependent response in TER and Ccl to cholesterol depletion

Page 18: Impedance Spectroscopy for Toxicity Testing Martin Dominguez Cronus  Technologies Ltd.

Testing the Toxicity of NanoparticlesExposure of MDCK cells to different types of particles

+ Triton X100

ControlControl

Carbon Black*

Carbon Black*

ZrO2*

ZrO2*

ZnO - NP*

ZnO - NP*

+ Particles

* 10 µg/cm²

0

20

40

60

80

100

120

140

160

180

0 5 10 15 20 25

Time (h)

TER

(%)

Data courtesy of A. Kroll, J. Schnekenburger, University of Münster, Germany

Page 19: Impedance Spectroscopy for Toxicity Testing Martin Dominguez Cronus  Technologies Ltd.

Testing the Toxicity of NanoparticlesExposure of NRK52E cells to ZnO

ControlControl

ZnO*

ZnO*

ZnO*

ZnO NP*

ZnO NP*

ZnO NP*

* 10 µg/cm²

0

20

40

60

80

100

120

0 5 10 15 20 25

TER

(%)

+ ZnO + Triton X100

Time (h)

Data courtesy of A. Kroll, J. Schnekenburger, University of Münster, Germany

Page 20: Impedance Spectroscopy for Toxicity Testing Martin Dominguez Cronus  Technologies Ltd.

cellZscopeAutomated cell monitoring device