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Cellular ATP content during and after calcium electroporation evaluated by fluorescence microscopy

STSM by Emilie Louise Hansen, Scholarship Student Researcher

Period of mission: from 01/04/2014 to 30/04/2014

Home institution: Department of Oncology, Copenhagen University Hospital Herlev, 2730 Herlev, Denmark

Host institution: Frank Reidy Research Center for Bioelectrics, Norfolk VA, USA

Calcium electroporation is a method used to increase intracellular calcium uptake in cells through permeabilization of the cell membrane by application of electrical pulses. At the Frank Reidy Research center for Bioelectrics it was possible to use scanning confocal imaging via a Leica TCS SP8 microscope for investigation of the hypothesis that ATP depletion caused by calcium electroporation could be partially responsible for inducing necrosis in cancer cells. We used quinacrine as a marker for ATP and imaged the fluorescence in U937 human leukemia cell line 15, 30 and 60 minutes after treatment. Permeabilization marked using propidium iodide was delayed and more irregular after calcium-electroporation compared to electroporation. The quinacrine fluorescence was lower in the electroporated and calcium-electroporated samples than in the controls 15 and 30 minutes after treatment, the fluorescence being lowest for the calcium-electroporated sample. Calcium electroporation caused a short-term decrease in quinacrine fluorescence, which may indicate ATP depletion, and significantly decreased cell viability after 24 and 48 hours. The decrease in quinacrine fluorescence intensity after 1 hour in both electroporated and non-electroporated samples with or without added calcium should be further investigated. More systematic investigations to follow these should use more specific ATP indicators, such as luminescence from luciferin-luciferase.

Publication(s) originating from this mission

Journal Plos One
Impact Factor (according to ISI Thomson) 3,234
Authors Emilie L. Hansen, Esin B. Sozer, Stefania Romeo, Stine K. Frandsen, Paul T. Vernier, Julie Gehl
Title Dose-dependent ATP depletion and Cancer Cell Death following Calcium Electroporation, Relative Effect of Calcium Concentration and Electric Field Strength
Year 2015
Issue 10 (4)
Pages e0122973
Status Published

Conference abstract(s) originating from this mission

Publication IFMBE Proceedings of the 1st World Congress on Electroporation and Pulsed Electric Fields in Biology, Medicine and Food & Environmental Technologies
Publisher Tomaz J, Kramar P (ed), 1st edn. Springer-Verlag, Singapur
Authors Stefania Romeo, Emilie L. Hansen, Stine K. Frandsen, Julie Gehl
Title Electroporation of a Bladder Cancer Cell Line in Presence of Calcium: Efficacy Dependence on Electric Field Strength and Calcium Concentration
Year 2015
Issue 53
Pages 151-154
Status Published

Oral presentation(s) originating from this mission

Event 1st World Congress on Electroporation and Pulsed Electric Fields in Biology, Medicine and Food & Environmental Technologies
Location Portorož/Slovenia
Period 06/09 - 10/09/2015
Authors Stefania Romeo, Emilie L. Hansen, Stine K. Frandsen, Julie Gehl
Title Electroporation of a Bladder Cancer Cell Line in Presence of Calcium: Efficacy Dependence on Electric Field Strength and Calcium Concentration

 


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