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

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Title: In situ evaluation of single-cell lysis by cytosol extraction observation through fluorescence decay and dielectrophoretic trapping time
Authors: Mernier, G.1 guillaume.mernier@epfl.ch, Majocchi, S.2, Mermod, N.2, Renaud, P.1
Source: Sensors & Actuators B: Chemical. May2012, Vol. 166-167, p907-912. 6p.
Subjects: Lysis, Cytosol, Fluorescence, Dielectrophoresis, Electric fields, Scientific observation, Diffusion
Abstract: 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. [Copyright &y& Elsevier]
Copyright of Sensors & Actuators B: Chemical is the property of Elsevier B.V. and its content may not be copied or emailed to multiple sites without the copyright holder's express written permission. Additionally, content may not be used with any artificial intelligence tools or machine learning technologies. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)
Database: Engineering Source
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Header DbId: egs
DbLabel: Engineering Source
An: 75450347
AccessLevel: 6
PubType: Academic Journal
PubTypeId: academicJournal
PreciseRelevancyScore: 0
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  Data: In situ evaluation of single-cell lysis by cytosol extraction observation through fluorescence decay and dielectrophoretic trapping time
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  Data: <searchLink fieldCode="AR" term="%22Mernier%2C+G%2E%22">Mernier, G.</searchLink><relatesTo>1</relatesTo><i> guillaume.mernier@epfl.ch</i><br /><searchLink fieldCode="AR" term="%22Majocchi%2C+S%2E%22">Majocchi, S.</searchLink><relatesTo>2</relatesTo><br /><searchLink fieldCode="AR" term="%22Mermod%2C+N%2E%22">Mermod, N.</searchLink><relatesTo>2</relatesTo><br /><searchLink fieldCode="AR" term="%22Renaud%2C+P%2E%22">Renaud, P.</searchLink><relatesTo>1</relatesTo>
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  Data: <searchLink fieldCode="JN" term="%22Sensors+%26+Actuators+B%3A+Chemical%22">Sensors & Actuators B: Chemical</searchLink>. May2012, Vol. 166-167, p907-912. 6p.
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  Data: <searchLink fieldCode="DE" term="%22Lysis%22">Lysis</searchLink><br /><searchLink fieldCode="DE" term="%22Cytosol%22">Cytosol</searchLink><br /><searchLink fieldCode="DE" term="%22Fluorescence%22">Fluorescence</searchLink><br /><searchLink fieldCode="DE" term="%22Dielectrophoresis%22">Dielectrophoresis</searchLink><br /><searchLink fieldCode="DE" term="%22Electric+fields%22">Electric fields</searchLink><br /><searchLink fieldCode="DE" term="%22Scientific+observation%22">Scientific observation</searchLink><br /><searchLink fieldCode="DE" term="%22Diffusion%22">Diffusion</searchLink>
– Name: Abstract
  Label: Abstract
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  Data: 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. [Copyright &y& Elsevier]
– Name: AbstractSuppliedCopyright
  Label:
  Group: Ab
  Data: <i>Copyright of Sensors & Actuators B: Chemical is the property of Elsevier B.V. and its content may not be copied or emailed to multiple sites without the copyright holder's express written permission. Additionally, content may not be used with any artificial intelligence tools or machine learning technologies. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract.</i> (Copyright applies to all Abstracts.)
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        Value: 10.1016/j.snb.2012.03.057
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        Text: English
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      – SubjectFull: Dielectrophoresis
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      – SubjectFull: Diffusion
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              Text: May2012
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