In situ evaluation of single-cell lysis by cytosol extraction observation through fluorescence decay and dielectrophoretic trapping time

Details

Serval ID
serval:BIB_060EB126E499
Type
Article: article from journal or magazin.
Collection
Publications
Title
In situ evaluation of single-cell lysis by cytosol extraction observation through fluorescence decay and dielectrophoretic trapping time
Journal
Sensors and Actuators B-Chemical
Author(s)
Mernier G., Majocchi S., Mermod N., Renaud P.
ISSN
0925-4005
Publication state
Published
Issued date
2012
Volume
166
Pages
907-912
Language
english
Abstract
We present a new method for lysis of single cells in continuous flow, where cells are sequentially trapped, lysed and released in an automatic process. Using optimized frequencies, dielectrophoretic trapping allows exposing cells in a reproducible way to high electrical fields for long durations, thereby giving good control on the lysis parameters. In situ evaluation of cytosol extraction on single cells has been studied for Chinese hamster ovary (CHO) cells through out-diffusion of fluorescent molecules for different voltage amplitudes. A diffusion model is proposed to correlate this out-diffusion to the total area of the created pores, which is dependent on the potential drop across the cell membrane and enables evaluation of the total pore area in the membrane. The dielectrophoretic trapping is no longer effective after lysis because of the reduced conductivity inside the cells, leading to cell release. The trapping time is linked to the time required for cytosol extraction and can thus provide additional validation of the effective cytosol extraction for non-fluorescent cells. Furthermore, the application of one single voltage for both trapping and lysis provides a fully automatic process including cell trapping, lysis, and release, allowing operating the device in continuous flow without human intervention.
Keywords
Cell lysis, Liquid electrodes, Dielectrophoretic trapping, Electroporation
Web of science
Create date
30/01/2013 11:30
Last modification date
20/08/2019 13:28
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