<i>MicroPrep</i>: Chip‐based dielectrophoretic purification of mitochondria

Moschallski, Meike; Hausmann, Monika; Posch, Anton; Paulus, Aran; Kunz, Nancy; Duong, Ta Nguyen Binh; Angres, Brigitte; Fuchsberger, Kai; Steuer, Heiko; Stoll, Dieter; Werner, S.; Hagmeyer, Britta; Stelzle, Martin

doi:10.1002/elps.201000097

Cited by 25 publications

(21 citation statements)

References 70 publications

(40 reference statements)

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“…[7][8][9] The technique has been used to characterize and manipulate a number of biological particles ranging from micrometer-size cells and bacteria to spores and viruses of nanometer dimensions. [10][11][12] The latest development of the technique and the scope of its applications have been recently compiled in two comprehensive reviews. 9,13 The work presented here demonstrates for the first time the DEP collection and repulsion of 16S and 23S subunits of E. coli rRNA using microelectrodes.…”

Section: Introductionmentioning

confidence: 99%

Dielectrophoretic manipulation of ribosomal RNA

et al. 2011

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The manipulation of ribosomal RNA ͑rRNA͒ extracted from E. coli cells by dielectrophoresis ͑DEP͒ has been demonstrated over the range of 3 kHz-50 MHz using interdigitated microelectrodes. Quantitative measurement using total internal reflection fluorescence microscopy of the time dependent collection indicated a positive DEP response characterized by a plateau between 3 kHz and 1 MHz followed by a decrease in response at higher frequencies. Negative DEP was observed above 9 MHz. The positive DEP response below 1 MHz is described by the ClausiusMossotti model and corresponds to an induced dipole moment of 3300 D with a polarizability of 7.8ϫ 10 −32 F m 2 . The negative DEP response above 9 MHz indicates that the rRNA molecules exhibit a net moment of Ϫ250 D, to give an effective permittivity value of 78.5 0 , close to that of the aqueous suspending medium, and a relatively small surface conductance value of ϳ0.1 nS. This suggests that our rRNA samples have a fairly open structure accessible to the surrounding water molecules, with counterions strongly bound to the charged phosphate groups in the rRNA backbone. These results are the first demonstration of DEP for fast capture and release of rRNA units, opening new opportunities for rRNA-based biosensing devices.

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Section: Introductionmentioning

confidence: 99%

Dielectrophoretic manipulation of ribosomal RNA

et al. 2011

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“…This rationale could generally explain why the work presented by Krishnan et al 16 was not carried out in a continuous manner and the process usually required about 20 min to complete. To address this issue, several DEP devices have been reported to generate three-dimensional (3D) electric field by using 3D or sidewall metallic electrodes such as double planar electrodes, 8,18 heavy doped silicon, 23 pyrolyzed SU-8 photoresist, 24 and electroplated gold. 25 Nevertheless, because their fabrication materials are mainly glass and silicon, liquid leakage is a problem, leading to complicated device assemblies.…”

Section: Nuttawut Lewpiriyawong and Chun Yang A)mentioning

confidence: 99%

AC-dielectrophoretic characterization and separation of submicron and micron particles using sidewall AgPDMS electrodes

Lewpiriyawong

Yang

2012

Biomicrofluidics

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The recent development of microfluidic "lab on a chip" devices requires the need to continuously separate submicron particles. Here, we present a PDMS microfluidic device with sidewall conducting PDMS (AgPDMS) composite electrodes capable of separating submicron particles in hydrodynamic flow. In particular, the device can service dual functions. First, the AgPDMS composite electrodes embedded in a sidewall of the device channel allow for performing AC-dielectrophoretic (DEP) characterization through direct microscopic observation of particle behavior. Characterization experiments are carried out for numerous parameters including particle size, medium conductivity, and AC field frequency to reveal important dielectrophoresis DEP information in terms of the crossover frequency and positive/negative DEP behavior under specific frequencies. Second, the device offers an advantage that sidewall AgPDMS composite electrodes can produce strong DEP effects throughout the entire channel height, and thus the robustness of the on-chip particle separation is demonstrated for continuous separation in a flowing mixture of 0.5 and 5 μm particles with 100% separation efficiency.

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“…Thirdly, Joule heating due to electric fields across a conducting medium will cause temperature increase, which has to be kept within limits still safe for biological particles. The dielectrophoretic separation technique developed for this purpose is schematically depicted in Fig.2 A 87, 88 , 89 . When electrodes are energized by a high frequency voltage, an inhomogeneous electric field reaching across the channel is generated giving rise to a repulsive dielectrophoretic force, F DEP according to…”

Section: B C Amentioning

confidence: 99%

From bioseparation to artificial micro-organs: microfluidic chip based particle manipulation techniques

Stelzle

2010

SPIE Proceedings

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Microfluidic device technology provides unique physical phenomena which are not available in the macroscopic world. These may be exploited towards a diverse array of applications in biotechnology and biomedicine ranging from bioseparation of particulate samples to the assembly of cells into structures that resemble the smallest functional unit of an organ. In this paper a general overview of chip-based particle manipulation and separation is given. In the state of the art electric, magnetic, optical and gravitational field effects are utilized. Also, mechanical obstacles often in combination with force fields and laminar flow are employed to achieve separation of particles or molecules. In addition, three applications based on dielectrophoretic forces for particle manipulation in microfluidic systems are discussed in more detail. Firstly, a virus assay is demonstrated. There, antibody-loaded microbeads are used to bind virus particles from a sample and subsequently are accumulated to form a pico-liter sized aggregate located at a predefined position in the chip thus enabling highly sensitive fluorescence detection. Secondly, subcellular fractionation of mitochondria from cell homogenate yields pure samples as was demonstrated by Western Blot and 2D PAGE analysis. Robust long-term operation with complex cell homogenate samples while avoiding electrode fouling is achieved by a set of dedicated technical means. Finally, a chip intended for the dielectrophoretic assembly of hepatocytes and endothelial cells into a structure resembling a liver sinusoid is presented. Such "artificial micro organs" are envisioned as substance screening test systems providing significantly higher predictability with respect to the in vivo response towards a substance under test.

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MicroPrep: Chip‐based dielectrophoretic purification of mitochondria

Cited by 25 publications

References 70 publications

Dielectrophoretic manipulation of ribosomal RNA

Dielectrophoretic manipulation of ribosomal RNA

AC-dielectrophoretic characterization and separation of submicron and micron particles using sidewall AgPDMS electrodes

From bioseparation to artificial micro-organs: microfluidic chip based particle manipulation techniques

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