Semifluorinated Alkylphosphonic Acids Form High-Quality Self-Assembled Monolayers on Ag-Coated Yttrium Barium Copper Oxide Tapes and Enable Filamentization of the Tapes by Microcontact Printing.

A custom-designed semifluorinated phosphonic acid, (9,9,10,10,11,11,12,12,13,13,14,14,15,15,16,16,16-heptadecafluorohexadecyl)phosphonic acid (F8H8PA), and a normal hexadecylphosphonic acid (H16PA) were synthesized and used to generate self-assembled monolayers (SAMs) on commercially available yttrium barium copper oxide (YBCO) tapes. In this study, we wished to evaluate the effectiveness of these monolayer films as coatings for selectively etching YBCO. Initial films formed by solution deposition and manual stamping using a non-patterned polydimethylsiloxane stamp allowed for a comparison of the film-formation characteristics. The resulting monolayers were characterized by X-ray photoelectron spectroscopy (XPS), contact angle goniometry, and polarization modulation infrared reflection absorption spectroscopy (PM-IRRAS). To prepare line-patterned (filamentized) YBCO tapes, standard microcontact printing (µ-CP) procedures were used. The stamped patterns on the YBCO tapes were characterized by scanning electron microscopy (SEM) before and after etching to confirm the effectiveness of the patterning process on the YBCO surface and energy-dispersive X-ray spectroscopy (EDX) to obtain the atomic composition of the exposed interface.

Novel Silicon Titanium Diboride Micropatterned Substrates for Cellular Patterning.

Both hard material photolithography and soft lithography are widely used for patterned cell culture. Soft lithography techniques enable bioactive molecule incorporation, however complex surface modifications are required to introduce specific ligands or proteins in conventional photolithography. In this study, we demonstrate human umbilical vein cell (HUVEC) and adult bone marrow derived mesenchymal stem cell (MSC) patterning on titanium diboride (TiB2) layers deposited on silicon (Si) substrates by electron-beam evaporation and micropatterned using photolithography. Micropatterned cell growth specificity on geometric shapes of circle and/or lines is achieved via differential growth factors adsorption in the presence of heparin. Specifically, the deposited films of TiB2 showed increased stiffness, hardness, hydrophilicity and surface charge when compared to background Si. These substrates were found to be compatible with HUVEC and MSC viability, based on biomarker expression and RNA-sequence transcriptome analysis. Cell-type dependent, micropattern selective cell growth, such as contact guidance, alignment, and durotaxis, were observed. In addition, MSC clustering was achieved, enabling a three-dimensional (3D) aggregate based microenvironment during culture. This study clearly demonstrates the potential of microfabricated Si and TiB2 biomaterials for patterned cell culture in vitro, independent of any additional surface modification.

Mitochondrial membrane studies using impedance spectroscopy with parallel pH monitoring.

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.

Use of the radiofrequency-intermodulation distortion technique to investigate intrinsic nonlinearity at the electrode-electrolyte interface.

We report on investigations of nonlinear radiofrequency responses of electrolytes with Na(+) and Cl(-) ions placed within gold electrodes of a capacitor. The sample was part of a frequency-adjustable inductance-capacitance-resistance (LCR) parallel resonant circuit, and measurements were carried out using the two frequencies intermodulation distortion technique. We employed double layer model to analyze the observed nonlinearities and their dependence on ionic concentration. Electrode-electrolyte interface polarization was found to be a predominant cause of this intrinsic nonlinearity and to be dependent on electrolytic ion concentration. We also measured and calculated coefficients of resistive and capacitive components of the observed nonlinearity.

Reduction of electrode polarization capacitance in low-frequency impedance spectroscopy by using mesh electrodes.

Dielectric measurements of biological samples are obscured by electrode polarization, which at low frequencies dominates over the actual sample response. Reduction of this artifact is especially necessary in studying interactions of electric field with biological systems in the α-dispersion range. We developed a method to reduce the influence of electrode polarization by employing mesh instead of solid electrodes as sensing probes, thereby reducing the area of the double layer. The design decreases the electrode-electrolyte contact area by almost 40% while keeping the bulk sample capacitance the same. Interrogation electric fields away from the electrode surface and sensitivity are unaffected. Electrodes were microfabricated (600µm×50µm, spacing of 100µm) with and without mesh holes 7.5µm×7.5µm in size. Simulations of electric field performed using Comsol Multiphysics showed non-uniformity of the electric field within less than 1.5µm from the electrode surface, which encompasses the double layer region, but at greater distance the solid and mesh electrodes gave the same results. Mesh electrodes reduced capacitance measurements for water and KCl solutions of different concentrations at low frequencies (<10kHz), while higher frequency capacitance remained the same for both electrode types, confirming our hypothesis that this design leaves the electric field mainly unaffected. Impedance measurements at low frequencies for water and mice heart mitochondrial suspension were lower for mesh than for solid electrodes. Comsol simulations confirmed these results by showing that mesh electrodes have a greater charge density than solid electrodes, which affects conductance. These electrodes are being used for mitochondrial membrane potential studies.