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Real time kinetic flow cytometry measurements of cellular parameter changes evoked by nanosecond pulsed electric field
Author(s) -
Orbán Csaba,
PérezGarcía Esther,
Bajnok Anna,
McBean Gethin,
Toldi Gergely,
BlancoFernandez Alfonso
Publication year - 2016
Publication title -
cytometry part a
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.316
H-Index - 90
eISSN - 1552-4930
pISSN - 1552-4922
DOI - 10.1002/cyto.a.22838
Subject(s) - ionomycin , nanosecond , flow cytometry , chemistry , biophysics , electric field , intracellular , nanotechnology , cytometry , cell , materials science , microbiology and biotechnology , biochemistry , biology , physics , laser , quantum mechanics , optics
Nanosecond pulsed electric field (nsPEF) is a novel method to increase cell proliferation rate. The phenomenon is based on the microporation of cellular organelles and membranes. However, we have limited information on the effects of nsPEF on cell physiology. Several studies have attempted to describe the effects of this process, however no real time measurements have been conducted to date. In this study we designed a model system which allows the measurement of cellular processes before, during and after nsPEF treatment in real time. The system employs a Vabrema Mitoplicator TM nsPEF field generating instrument connected to a BD Accuri C6 cytometer with a silicon tube led through a peristaltic pump. This model system was applied to observe the effects of nsPEF in mammalian C6 glioblastoma (C6 glioma) and HEK‐293 cell lines. Viability (using DRAQ7 dye), intracellular calcium levels (using Fluo‐4 dye) and scatter characteristics were measured in a kinetic manner. Data were analyzed using the FACSKin software. The viability and morphology of the investigated cells was not altered upon nsPEF treatment. The response of HEK‐293 cells to ionomycin as positive control was significantly lower in the nsPEF treated samples compared to non‐treated cells. This difference was not observed in C6 cells. FSC and SSC values were not altered significantly by the nsPEF treatment. Our results indicate that this model system is capable of reliably investigating the effects of nsPEF on cellular processes in real time. © 2016 International Society for Advancement of Cytometry