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

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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

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

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Understanding Impedance SpectraAn increase in the capacitance CCL narrows the plateau

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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

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

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

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

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0

20

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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

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

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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

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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

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cellZscopeAutomated cell monitoring device