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Published online by Cambridge University Press: 31 January 2011
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.