We designed planar electrodes, for dielectrophoretic manipulation of single-walled carbon nanotubes (SWNTs), built as metal-oxide-semiconductor nanogap capacitors with common substrate and oxide thicknesses of 17 and 150 nm. Such design generates high electric fields (10(9) V m(-1)) and also the fringing field is curved due to the conducting substrate, unlike fields generated by conventionally used planar electrodes. Scanning electron microscopy images showed SWNTs aligned parallel and perpendicular to the electrodes. Raman spectroscopic mapping was used to produce separate images of the metallic (m-SWNT) and semiconducting (s-SWNT) nanotube density distributions. As expected, parallel alignment of the m-SWNTs with the E-field was found; however, also a perpendicular alignment of s-SWNTs was observed. Such orthogonal alignment of s-SWNTs is a rare observation and has not been experimentally reported before in detail with Raman images. Due to the unique electrode design, we were able to obtain substantial separation of m-SWNTs and s-SWNTs. Numerical modeling of the electric field factor of the dielectrophoresis force was done, and it matched perfectly with the experimental results. The orthogonal alignment of s-SWNTs results from comparable values of parallel and perpendicular polarizability to the nanotube axis.
A biological microelectromechanical system (BioMEMS) device was designed to study complementary mitochondrial parameters important in mitochondrial dysfunction studies. Mitochondrial dysfunction has been linked to many diseases, including diabetes, obesity, heart failure and aging, as these organelles play a critical role in energy generation, cell signaling and apoptosis. The synthesis of ATP is driven by the electrical potential across the inner mitochondrial membrane and by the pH difference due to proton flux across it. We have developed a tool to study the ionic activity of the mitochondria in parallel with dielectric measurements (impedance spectroscopy) to gain a better understanding of the properties of the mitochondrial membrane. This BioMEMS chip includes: 1) electrodes for impedance studies of mitochondria designed as two- and four-probe structures for optimized operation over a wide frequency range and 2) ion-sensitive field effect transistors for proton studies of the electron transport chain and for possible monitoring other ions such as sodium, potassium and calcium. We have used uncouplers to depolarize the mitochondrial membrane and disrupt the ionic balance. Dielectric spectroscopy responded with a corresponding increase in impedance values pointing at changes in mitochondrial membrane potential. An electrical model was used to describe mitochondrial sample’s complex impedance frequency dependencies and the contribution of the membrane to overall impedance changes. The results prove that dielectric spectroscopy can be used as a tool for membrane potential studies. It can be concluded that studies of the electrochemical parameters associated with mitochondrial bioenergetics may render significant information on various abnormalities attributable to these organelles.
We developed a BioMEMS device to study cell- mitochondrial physiological functionalities. The pathogenesis of many diseases including obesity, diabetes, heart failure as well as aging has been linked to functional defects of mitochondria. This is understandable as the mitochondria produces up to 90% of ATP, and plays a critical role in cell signaling and apoptosis. The synthesis of ATP is determined by the electrical potential across the inner mitochondrial membrane (IMM) and by the pH difference due to proton flux across it. Therefore, electrical characterization by E-fields with complementary chemical testing was used here. Mitochondrial ion channels present in the IMM control specific ion fluxes, and maintain ion homeostasis, matrix volume, IMM potential etc and thus serve a central role in cell growth and death related processes. Defects in ion channels (Channelopathies) are being attributed to many diseases like cancer, neurodegeneration, etc. Complete physiological roles of various ion channels and their interactions are still unknown, hindering the development of targeted therapeutic agents. The BioMEMS device was fabricated as an SU-8 based microfluidic system with gold electrodes on SiO2/Si wafers for electromagnetic interrogation. Ion Sensitive Field Effect Transistors (ISFETs) were incorporated for proton studies important in electron transport chain, together with monitoring Na+, K+, Ca++ions for ion channel studies. ISFETs are chemically sensitive MOSFET devices, their threshold voltage is directly proportional to the electrolytic H+ ion variation. These ISFETs (sensitivity ˜55 mV/pH for H+) were further realized as specific ion sensitive CHEMFETs by depositing a poly-HEMA layer sandwiched between the gate and a final specific ion sensitive membrane. Electrodes for dielectric spectroscopy studies of mitochondria were designed as 2- and 4-probe structures for optimized operation over a wide frequency range. In addition, to limit polarization effects (which masks actual impedance for high conductivity solutions at low frequencies), a 4-electrode set-up with unique meshed pickup electrodes (7.5×7.5 μm2 loops with 4 μm wires) was fabricated. An electrical model was developed for the mitochondrial sample, and its frequency response correlated with impedance spectroscopy experiments of sarcolemmal mitochondria. Using the mesh electrode structure, we obtained a reduction of 83.28% in impedance at 200 Hz. COMSOL simulations of selected electrical structures in this sensor were compared with experimental results to better understand the physical system. The simultaneous measurement of membrane potential, ion concentrations and pH would enhance diagnostics and studies of mitochondrial diseases.
Nano-gap metal oxide semiconductor (MOS) capacitors were studied to evaluate their limitations in applications of dielectric spectroscopy in living cells. The purpose was to optimize the design of a transducer to avoid interfacial polarization at the electrodes. Silicon IC technology was selected for designing processes in which we could limit electric double layer impedance by precisely controlling dielectric thickness of the capacitors in the range of 17 to 150 nm. The working capacitance was defined by lateral oxide etching of capacitor structures of various configuration to ensure high perimeter to area ratio. Highly doped n+ polysilicon and n+ implanted Si substrate were acting as capacitor's electrodes. Restrictions known from CMOS circuits regarding oxide leakage current, which depends on geometry and increases with the gate area were taken into account. To allow for testing cells (yeasts), which have larger dimensions than nano structures it was necessary to include cell manipulation using dielectrophoresis (DEP). Entrapment of cells at the electrode perimeter preceded electrical measurements. Our focus in analyses was on the frequency dependence of impedance parameters.
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