Mechanism of Cell Lysis in Microfluidic Channel With Integrated Nanocomposite Electrodes

Author(s):  
Madhusmita Mishra ◽  
Anil Krishna Koduri ◽  
Aman Chandra ◽  
D. Roy Mahapatra ◽  
G. M. Hegde

This paper reports on the characterization of an integrated micro-fluidic platform for controlled electrical lysis of biological cells and subsequent extraction of intracellular biomolecules. The proposed methodology is capable of high throughput electrical cell lysis facilitated by nano-composite coated electrodes. The nano-composites are synthesized using Carbon Nanotube and ZnO nanorod dispersion in polymer. Bacterial cells are used to demonstrate the lysis performance of these nanocomposite electrodes. Investigation of electrical lysis in the microchannel is carried out under different parameters, one with continuous DC application and the other under DC biased AC electric field. Lysis in DC field is dependent on optimal field strength and governed by the cell type. By introducing the AC electrical field, the electrokinetics is controlled to prevent cell clogging in the micro-channel and ensure uniform cell dispersion and lysis. Lysis mechanism is analyzed with time-resolved fluorescence imaging which reveal the time scale of electrical lysis and explain the dynamic behavior of GFP-expressing E. coli cells under the electric field induced by nanocomposite electrodes. The DNA and protein samples extracted after lysis are compared with those obtained from a conventional chemical lysis method by using a UV–Visible spectroscopy and fluorimetry. The paper also focuses on the mechanistic understanding of the nano-composite coating material and the film thickness on the leakage charge densities which lead to differential lysis efficiency.

Lab on a Chip ◽  
2008 ◽  
Vol 8 (3) ◽  
pp. 408 ◽  
Author(s):  
Pedro A. Quinto-Su ◽  
Hsuan-Hong Lai ◽  
Helen H. Yoon ◽  
Christopher E. Sims ◽  
Nancy L. Allbritton ◽  
...  

Author(s):  
Kendra V. Sharp ◽  
Scott M. Davison ◽  
Shahrzad H. Yazdi

Work with dc electrokinetics has demonstrated that is works well for bulk transport of fluid an particles. However, it is difficult to achieve control of individual or groups of particles. This paper investigate the use of induced-charge electroosmosis (ICEO) as a means of providing control over particles within bulk dc electroosmotic flow. ICEO flow develops when an electric double layer is induced by an applied electric field at the surface of a conducting object. Here conducting posts are positioned in a microfluidic channel and ICEO flow develops around them due to an applied ac electric field. A dc electric field is applied across the length of the channel to induce electroosmotic flow past the ICEO region. Around one arrangement of posts the ac and dc flow fields combine to produce a region of recirculation which could be useful for holding a particle or particles within a fixed region of the channel. An alternative arrangement of posts functions to focus the flow into the center of the channel. A numerical model of the system is developed and used to explore means of adapting the ICEO flows to many situations. A method for fabricating a microfluidic system for ICEO flows is presented.


Author(s):  
Christopher Church ◽  
Junjie Zhu ◽  
Guohui George Huang ◽  
Gaoyan Wang ◽  
Tzuen-Rong Jeremy Tzeng ◽  
...  

Cell lysis is a necessary step in the analysis of intracellular contents. It has been recently demonstrated in microfluidic devices using four methods: chemical lysis, mechanical lysis, thermal lysis, and electrical lysis [1]. The locally high electric fields needed for electrical lysis have been achieved using micro-electrodes and micro-constrictions for pulsed and continuous DC electric fields, respectively. However, since the two determining factors of electrical lysis are field strength and exposure time, opposing pressure-driven flow must often be used in pure DC lysis to reduce the velocity of the cells and to ensure the cells spend sufficient time in the high electric field region [1,2]. Using DC-biased AC fields can easily fulfill these requirements as only the DC component contributes to cell electrokinetic transport. Prior to lysis, cell concentration can be increased by trapping using dielectrophoresis (DEP), which may occur with either DC or DC-biased AC electric fields [3,4]. This operation is useful in cases where the cell supply is limited or when the cell concentration is too low in general. In this work, red blood cells are used to demonstrate the smooth switching between electrical lysing and trapping in a microchannel constriction. The transition between lysis and trapping is realized by tuning the DC component in a DC-biased AC electric field.


2018 ◽  
Vol 189 (02) ◽  
pp. 187-194 ◽  
Author(s):  
Nikita V. Marchenkov ◽  
Anton G. Kulikov ◽  
Ivan I. Atknin ◽  
Arsen A. Petrenko ◽  
Alexander E. Blagov ◽  
...  

2015 ◽  
Vol 108 ◽  
pp. 183-191 ◽  
Author(s):  
Reza Riahifar ◽  
Babak Raissi ◽  
Cyrus Zamani ◽  
Ehsan Marzbanrad

2014 ◽  
Vol 99 ◽  
pp. 160-163 ◽  
Author(s):  
Hiroshi Kimura ◽  
Mao Ueno ◽  
Shinya Takahashi ◽  
Akira Tsuchida ◽  
Keiichi Kurosaka

Antibiotics ◽  
2021 ◽  
Vol 10 (3) ◽  
pp. 341
Author(s):  
Katharina Hoenes ◽  
Richard Bauer ◽  
Barbara Spellerberg ◽  
Martin Hessling

Interest in visible light irradiation as a microbial inactivation method has widely increased due to multiple possible applications. Resistance development is considered unlikely, because of the multi-target mechanism, based on the induction of reactive oxygen species by wavelength specific photosensitizers. However, the affected targets are still not completely identified. We investigated membrane integrity with the fluorescence staining kit LIVE/DEAD® BacLight™ on a Gram positive and a Gram negative bacterial species, irradiating Staphylococcus carnosus and Pseudomonas fluorescens with 405 nm and 450 nm. To exclude the generation of viable but nonculturable (VBNC) bacterial cells, we applied an ATP test, measuring the loss of vitality. Pronounced uptake of propidium iodide was only observed in Pseudomonas fluorescens at 405 nm. Transmission electron micrographs revealed no obvious differences between irradiated samples and controls, especially no indication of an increased bacterial cell lysis could be observed. Based on our results and previous literature, we suggest that visible light photoinactivation does not lead to rapid bacterial cell lysis or disruption. However, functional loss of membrane integrity due to depolarization or inactivation of membrane proteins may occur. Decomposition of the bacterial envelope following cell death might be responsible for observations of intracellular component leakage.


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