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Multiple nanosecond electric pulses increase the number but not the size of long-lived nanopores in the cell membrane
- Source :
- Biochimica et Biophysica Acta (BBA) - Biomembranes. 1848(4):958-966
- Publication Year :
- 2015
- Publisher :
- Elsevier BV, 2015.
-
Abstract
- Exposure to intense, nanosecond-duration electric pulses (nsEP) opens small but long-lived pores in the plasma membrane. We quantified the cell uptake of two membrane integrity marker dyes, YO-PRO-1 (YP) and propidium (Pr), in order to test whether the pore size is affected by the number of nsEP. The fluorescence of the dyes was calibrated against their concentrations by confocal imaging of stained homogenates of the cells. The calibrations revealed a two-phase dependence of Pr emission on the concentration (with a slower rise at < 4 µM), and a linear dependence for YP. CHO cells were exposed to nsEP trains (1 to 100 pulses, 60 ns, 13.2 kV/cm, 10 Hz) with Pr and YP in the medium, and the uptake of the dyes was monitored by time-lapse imaging for 3 min. Even a single nsEP triggered a modest but detectable entry of both dyes, which increased linearly when more pulses were applied. The influx of Pr per pulse was constant and independent of the pulse number. The influx of YP per pulse was highest with 1- and 2-pulse exposures, decreasing to about twice the Pr level for trains from 5 to 100 pulses. The constant YP/Pr influx ratio for trains of 5 to 100 pulses suggests that increasing the number of pulses permeabilizes cells to a greater extent by increasing the pore number and not the pore diameter.
- Subjects :
- Cell Membrane Permeability
Membrane permeability
Analytical chemistry
Biophysics
CHO Cells
02 engineering and technology
Time-Lapse Imaging
Biochemistry
Article
Cell membrane
03 medical and health sciences
Nanopores
Cricetulus
Electromagnetic Fields
Cricetinae
medicine
Animals
Pulse
030304 developmental biology
Benzoxazoles
0303 health sciences
Nanosecond pulses
Chemistry
Pulse (signal processing)
Quinolinium Compounds
Electroporation
Cell Membrane
Cell Biology
Nanosecond
021001 nanoscience & nanotechnology
Fluorescence
Electric Stimulation
Nanopore
medicine.anatomical_structure
Membrane
Electropermeabilization
0210 nano-technology
nsEP
Propidium
Subjects
Details
- ISSN :
- 00052736
- Volume :
- 1848
- Issue :
- 4
- Database :
- OpenAIRE
- Journal :
- Biochimica et Biophysica Acta (BBA) - Biomembranes
- Accession number :
- edsair.doi.dedup.....5ae418f71f99cbefc90d28ba3ce2c994
- Full Text :
- https://doi.org/10.1016/j.bbamem.2014.12.026