<abstract xmlns="http://www.w3.org/1999/xhtml"><p> Electroporation is an electric field induced phenomenon occurring when the permeability of the cell membrane is increased due to the excess of critical transmembrane potential. Fluorescent dye assays are frequently used for evaluation of the permeabilization rate, however, the protocols vary, which negatively affects the repeatability of the results. In this work we have designed experiments to investigate the protocols and threshold concentrations of the Propidium Iodide (PI) and YO-PRO-1 (YP) fluorescent dyes for evaluation of mammalian cell permeabilization induced by electroporation. The Sp2/0 mouse myeloma cells were used and the bursts of 100 μs × 8 electrical pulses of 0.8-2 kV/cm were applied. It has been shown that the dye concentration has an influence on the detectable permeabilization, and the concentrations below 30 μM for PI and 1 μM for YP should be avoided for measurement of electropermeabilization efficacy due to unreliable fluorescence signals. Further, based on the experimental data, the permeabilization curve for the Sp2/0 myeloma cells in the 0.8-2 kV/cm range has been presented.</p></abstract>

Electroporation is an electric field induced phenomenon occurring when the permeability of the cell membrane is increased due to the excess of critical transmembrane potential. Fluorescent dye assays are frequently used for evaluation of the permeabilization rate, however, the protocols vary, which negatively affects the repeatability of the results. In this work we have designed experiments to investigate the protocols and threshold concentrations of the Propidium Iodide (PI) and YO-PRO-1 (YP) fluorescent dyes for evaluation of mammalian cell permeabilization induced by electroporation. The Sp2/0 mouse myeloma cells were used and the bursts of 100 μs × 8 electrical pulses of 0.8-2 kV/cm were applied. It has been shown that the dye concentration has an influence on the detectable permeabilization, and the concentrations below 30 μM for PI and 1 μM for YP should be avoided for measurement of electropermeabilization efficacy due to unreliable fluorescence signals. Further, based on the experimental data, the permeabilization curve for the Sp2/0 myeloma cells in the 0.8-2 kV/cm range has been presented.

Measurement of Transient Permeability of Sp2/0 Myeloma Cells: Flow Cytometric Study

Institute of High Magnetic Fields, Faculty of Electronics, Vilnius Gediminas Technical University, Naugarduko g. 41, 03227, Vilnius,

State Research Institute Centre for Innovative Medicine, Department of Immunology, Santariškių g. 5, 08406, Vilnius,

Department of Microbiology and Biotechnology, Vilnius University, Sauletekio al. 9, 10221, Vilnius,

Department of Biodeterioration Research, Nature Research Centre, Akademijos str. 2, 08412, Vilnius,

Laboratory of Mycology, Nature Research Centre, Akademijos str. 2, 08412, Vilnius,

Measurement Science Review

This work is licensed under the Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 License.

Electroporation is an electric field induced phenomenon occurring when the permeability of the cell membrane is increased due to the excess of critical...

Measurement of Transient Permeability of Sp2/0 Myeloma Cells: Flow Cytometric Study

Publié en ligne: 13 déc. 2016

Pages: 300 - 304

Reçu: 09 sept. 2016

Accepté: 25 nov. 2016

DOI: https://doi.org/10.1515/msr-2016-0038

Mots clés
Electroporation, BD FACS Calibur, Amnis FlowSight, Sp2/0 myeloma cells

© by Vitalij Novickij

This work is licensed under the Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 License.

Measurement of Transient Permeability of Sp2/0 Myeloma Cells: Flow Cytometric Study

Publié en ligne: 13 déc. 2016

Pages: 300 - 304

Reçu: 09 sept. 2016

Accepté: 25 nov. 2016

DOI: https://doi.org/10.1515/msr-2016-0038

Mots clésElectroporation, BD FACS Calibur, Amnis FlowSight, Sp2/0 myeloma cells

© by Vitalij Novickij

This work is licensed under the Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 License.

Mots clés
Electroporation, BD FACS Calibur, Amnis FlowSight, Sp2/0 myeloma cells