Publication Types:

Application of Variational Principle to Form Reduced Fluid-Structure Interaction Models in Bifurcated Networks

A.S. Liberson, Y. Seyed Vahedein, D. A. Borkholder
Journal of Fluid Flow, Heat and Mass Transfer, Volume 4, August 2017, Pages 1-9
Publication year: 2017

Abstract – Reduced fluid-structure interaction models have received a considerable attention in recent years being the key component of hemodynamic modeling. A variety of models applying to specific physiological components such as arterial, venous and cerebrospinal fluid (CSF) circulatory systems have been developed based on different approaches. The purpose of this paper is to apply the general approach based on Hamilton’s variational principle to create a model for a viscous Newtonian Fluid – Structure Interaction (FSI) in a compliant bifurcated network. This approach provides the background for a correct formulation of reduced FSI models with an account for arbitrary nonlinear visco-elastic properties of compliant boundaries. The correct boundary conditions are specified at junctions, including the interface between 3D and 1D models. The hyperbolic properties of the derived mathematical model are analyzed and used, constructing a monotone finite volume numerical scheme, second-order accuracy in time and space. The computational algorithm is validated by comparison of numerical solutions with the exact manufactured solutions for an isolated compliant segment and a bifurcated structure. The accuracy of applied total variation diminishing (TVD) and Lax-Wendroff schemes are analyzed by comparison of numerical results to the available analytical smooth and discontinuous solutions, demonstrating a superior performance from the TVD algorithm.


Towards an Implantable, Low Flow Micropump That Uses No Power in the Blocked-Flow State

Dean G. Johnson, and David A. Borkholder
Micromachines 2016, 7, 99.
Publication year: 2016

Low flow rate micropumps play an increasingly important role in drug therapy research. Infusions to small biological structures and lab-on-a-chip applications require ultra-low flow rates and will benefit from the ability to expend no power in the blocked-flow state. Here we present a planar micropump based on gallium phase-change actuation that leverages expansion during solidification to occlude the flow channel in the off-power state. The presented four chamber peristaltic micropump was fabricated with a combination of Micro Electro Mechanical System (MEMS) techniques and additive manufacturing direct write technologies. The device is 7 mm × 13 mm × 1 mm (<100 mm3) with the flow channel and exterior coated with biocompatible Parylene-C, critical for implantable applications. Controllable pump rates from 18 to 104 nL/min were demonstrated, with 11.1 ± 0.35 nL pumped per actuation at an efficiency of 11 mJ/nL. The normally-closed state of the gallium actuator prevents flow and diffusion between the pump and the biological system or lab-on-a-chip, without consuming power. This is especially important for implanted applications with periodic drug delivery regimens.


Quantification of Hemodynamic Pulse Wave Velocity Based on a Thick Wall Multi-Layer Model for Blood Vessels

Jeffrey S. Lillie, Alexander S. Liberson, David A. Borkholder
Journal of Fluid Flow, Volume 3, 2016 - Pages 54-61. DOI: 10.11159/jffhmt.2016.007
Publication year: 2016

Pulse wave velocity (PWV) is an important index of arterial hemodynamics, which lays the foundation for continuous, noninvasive blood pressure automated monitoring. The goal of this paper is to examine the accuracy of PWV prediction based on a traditional homogeneous structural model for thin-walled arterial segments. In reality arteries are described as composite heterogeneous hyperelastic structures, where the thickness dimension cannot be considered small compared to the cross section size. In this paper we present a hemodynamic fluid – structure interaction model accounting for the variation of geometry and material properties in a radial direction. The model is suitable to account for the highly nonlinear orthotropic material undergoing finite deformation for each layer. Numerical analysis of single and two layer arterial segments shows that a single thick layer model provides sufficient accuracy to predict PWV. The dependence of PWV on pressure for three vessels of different thicknesses is compared against a traditional thin wall model of a membrane shell interacting with an incompressible fluid. The presented thick wall model provides greater accuracy in the prediction of PWV, and will be important for blood pressure estimation based on PWV measurements.


Photonic Sintering of Aerosol Jet Printed Lead Zirconate Titanate (PZT) Thick Films

Jing Ouyang, Denis Cormier, Scott A. Williams, David A. Borkholder
Journal of the American Ceramic Society, Volume 99, Pages 2569–2577, 2016. DOI: 10.1111/jace.14272
Publication year: 2016

Lead Zirconate Titanate (PZT) is a commonly used piezoelectric material due to its high piezoelectric response. We demonstrate a new method of printing and sintering micro-scale PZT films with low substrate temperature increase. Self-prepared PZT ink was Aerosol-Jet printed on stainless steel substrates. After drying for 2 h in vacuum at 200°C, the printed PZT films were divided into two groups. The first group was traditionally sintered, using a thermal process at 1000°C for 1 h in an Argon environment. The second group was photonically sintered using repetitive sub-msec pulses of high intensity broad spectrum light in an atmospheric environment. The highest measured substrate temperature during photonic sintering was 170.7°C, enabling processing on low melting point substrates. Ferroelectric measurements were performed with a low-frequency sinusoidal signal. The remanent polarization (Pr) and coercive field (Ec) for thermally sintered PZT film were 17.1 μC/cm2 and 6.3 kV/cm, respectively. The photonically sintered film had 32.4 μC/cm2 Pr and 6.7 kV/cm Ec. After poling the samples with 20 kV/cm electric field for 2 h at 150°C, the piezoelectric voltage constant (g33) was measured for the two film groups yielding −16.9 × 10−3 (V·m)·N−1 (thermally sintered) and −17.9 × 10−3 (V·m)·N−1(photonically sintered). Both factors indicate the PZT films were successfully sintered using both methods, with the photonically sintered material exhibiting superior electrical properties. To further validate photonic sintering of PZT on low melting point substrates, the process and measurements were repeated using a polyethylene terephthalate (PET) substrate. The measured Pr and Ec were 23.1 μC/cm2 and 5.1 kV/cm, respectively. The g33 was −17.3 × 10−3 (V·m)·N−1. Photonic sintering of thick film PZT directly on low melting point substrates eliminates the need for complex layer transfer processes often associated with flexible PZT transducers.



Improved Blood Pressure Prediction Using Systolic Flow Correction of Pulse Wave Velocity

Jeffrey S. Lillie, Alexander S. Liberson, David A. Borkholder
Cardiovascular Engineering and Technology, 2016, Pages 1-9.  DOI: 10.1007/s13239-016-0281-y
Publication year: 2016

Hypertension is a significant worldwide health issue. Continuous blood pressure monitoring is important for early detection of hypertension, and for improving treatment efficacy and compliance. Pulse wave velocity (PWV) has the potential to allow for a continuous blood pressure monitoring device; however published studies demonstrate significant variability in this correlation. In a recently presented physics-based mathematical model of PWV, flow velocity is additive to the classic pressure wave as estimated by arterial material properties, suggesting flow velocity correction may be important for cuff-less non-invasive blood pressure measures. The present study examined the impact of systolic flow correction of a measured PWV on blood pressure prediction accuracy using data from two published in vivo studies. Both studies examined the relationship between PWV and blood pressure under pharmacological manipulation, one in mongrel dogs and the other in healthy adult males. Systolic flow correction of the measured PWV improves the R2 correlation to blood pressure from 0.51 to 0.75 for the mongrel dog study, and 0.05 to 0.70 for the human subjects study. The results support the hypothesis that systolic flow correction is an essential element of non-invasive, cuff-less blood pressure estimation based on PWV measures.



A physics based approach to the pulse wave velocity prediction in compliant arterial segments

Alexander S. Liberson, Jeffrey S. Lillie, Steven W. Day, David A. Borkholder
Journal of Biomechanics, Volume 49, Issue 14, 2016, Pages 3460-3466.  DOI: 10.1016/j.jbiomech.2016.09.013
Publication year: 2016

Pulse wave velocity (PWV) quantification commonly serves as a highly robust prognostic parameter being used in a preventative cardiovascular therapy. Being dependent on arterial elastance, it can serve as a marker of cardiovascular risk. Since it is influenced by a blood pressure (BP), the pertaining theory can lay the foundation in developing a technique for noninvasive blood pressure measurement. Previous studies have reported application of PWV, measured noninvasively, for both the estimation of arterial compliance and blood pressure, based on simplified physical or statistical models. A new theoretical model for pulse wave propagation in a compliant arterial segment is presented within the framework of pseudo-elastic deformation of biological tissue undergoing finite deformation. An essential ingredient is the dependence of results on nonlinear aspects of the model: convective fluid phenomena, hyperelastic constitutive relation, large deformation and a longitudinal pre-stress load. An exact analytical solution for PWV is presented as a function of pressure, flow and pseudo-elastic orthotropic parameters. Results from our model are compared with published in-vivo PWV measurements under diverse physiological conditions. Contributions of each of the nonlinearities are analyzed. It was found that the totally nonlinear model achieves the best match with the experimental data. To retrieve individual vascular information of a patient, the inverse problem of hemodynamics is presented, calculating local orthotropic hyperelastic properties of the arterial wall. The proposed technique can be used for non-invasive assessment of arterial elastance, and blood pressure using direct measurement of PWV, with account of hyperelastic orthotropic properties.


The electric field strength in orifice-like nanopores of ultrathin membranes.

Getpreecharsawas J, McGrath J, Borkholder DA
Nanotechnology, 26(4) (2015): 045704. doi:10.1088/0957-4484/26/4/045704
Publication year: 2015

Here we show that the electric field inside an ultrathin membrane is weaker than conventional theory would predict, and that the reduced field is predictive of measured electroosmotic flow rates. Our theoretical analysis shows that the electric field inside a charged nanopore is affected by end effects and dependent on the Dukhin number Du when the pore length-to-diameter aspect ratio λ is less than 80 for Du ≪ 1 or 300 for Du ≫ 1. The electric field follows an unconventional scaling law; it no longer scales uniformly with the thickness of membrane, but with the local value of λ for each nanopore.



Pulse wave velocity prediction and compliance assessment in elastic arterial segments.

Lillie JS, Liberson AS, Mix D, Schwarz QS, Chandra A, Phillips DB, Day SW, Borkholder, DA
Cardiovascular Engineering and Technology, 6(1):49-58 (2015): DOI:10.1007/s13239-014-0202-x.
Publication year: 2015

Pressure wave velocity (PWV) is commonly used as a clinical marker of vascular elasticity. Recent studies have increased clinical interest in also analyzing the impact of heart rate (HR), blood pressure, and left ventricular ejection time (LVET) on PWV. In this article we focus on the development of a theoretical one-dimensional model and validation via direct measurement of the impact of ejection time and peak pressure on PWV using an in vitro hemodynamic simulator. A simple nonlinear traveling wave model was developed for a compliant thin-walled elastic tube filled with an incompressible fluid. This model accounts for the convective fluid phenomena, elastic vessel deformation, radial motion, and inertia of the wall. An exact analytical solution for PWV is presented which incorporates peak pressure, ejection time, ejection volume, and modulus of elasticity. To assess arterial compliance, the solution is introduced in an alternative form, explicitly determining compliance of the wall as a function of the other variables. The model predicts PWV in good agreement with the measured values with a maximum difference of 3.0%. The results indicate an inverse quadratic relationship (R^2=.99) between ejection time and PWV, with ejection time dominating the PWV shifts (12%) over those observed with changes in peak pressure (2%). Our modeling and validation results both explain and support the emerging evidence that, both in clinical practice and clinical research, cardiac systolic function related variables should be regularly taken into account when interpreting arterial function indices, namely PWV.


The Focal Index as a Singular Metric for Beamforming Effectiveness

Matthew Kelly, Vincent Amuso, David Eddins, and David A Borkholder
Journal of the Acoustical Society of America, 136.5 (2014): 2654-2664.
Publication year: 2014

Hard of hearing individuals frequently cite intelligibility problems in multi-talker environments. Microphone arrays performing time delay beamforming address conditions of poor signal-to-noise ratio by spatially filtering incoming sound. Existing beampattern metrics including peak side lobe level, integrated side lobe level, beamwidth, and planar directivity index fail to quantitatively capture all elements essential for improving speech intelligibility in multi- talker situations. The focal index (FI) was developed here to address these deficiencies. Simulations were performed to exemplify the robust nature of the FI and to demonstrate the utility of this metric for driving array parameter selection. Beampatterns were generated and the metrics were calculated and evaluated against the strict unidirectional requirements for the array. Array performance was assessed by human subjects in a speech recognition task with competing speech from multiple locations. Simulations of array output were presented under conditions differing in array sparsity. The resulting human subject data was used to demonstrate the linear relationship (R2 > 0.975) between speech-intelligibility-weighted FI (SII-FI) and the signal-to- noise ratio thresholds for 20% and 80% correct responses. Data indicate that the FI and SII-FI are robust singular metrics for determining the effectiveness of the array.


Numerical Solution for the Boussinesq Type Models with Application to Arterial Flow

Liberson A., Lillie J, Borkholder DA
Journal of Fluid Flow, 1 (2014)
Publication year: 2014

A family of classical Boussinesq system of nonlinear wave theory is presented in a form of conservative equations supplemented by Riemann solver, which is a fundamental block in the Godunov frame formulation of flow problems. The governing system simulates different physical phenomena, such as propagation of a small amplitude waves on a surface of water, or pulse wave propagation in compliant thin walled arterial systems. While the first model is for verification purpose only, pulse wave propagation through the junction of thin walled elastic branches is of primary interest. The inertial effects associated with transversal motion of the wall are introduced, affirming the dispersive nature of pulsating waves. The problem of accounting for branching and possible discontinuity of wall properties is addressed. Preliminary analysis is presented which leads to the correct jump conditions across bifurcated area. As a result, the model accurately describes the formation of transmission and reflection waves at bifurcation, including effects of discontinuities. The Riemann solver supplies the inter cell flux and junction fluxes, based on conservation of volume, momentum and energy, with account of losses associated with the flow turn angle at bifurcation. An implicit monotonic total variation diminishing (TVD) scheme, second order accurate in time and space has been applied for the analysis of solitary wave solution in bifurcated arteries. Numerical results are in a good agreement with the known analytical and numerical solutions reported elsewhere. Based on direct computational analysis the inverse solution was obtained, calculating the local elastic properties of the arterial wall, using typical diagnostic measurements. Mathematical modelling presented in this work leads to a physiological understanding and interpretation of diagnostic measurements of the wave forms of a blood pressure, flow rate and an artery wall deflection.



Fetal QRS extraction from abdominal recordings via model-based signal processing and intelligent signal merging

Haghpanahi, Masoumeh, and David A. Borkholder
Physiological measurement 35.8 (2014): 1591.
Publication year: 2014

Noninvasive fetal ECG (fECG) monitoring has potential applications in diagnosing congenital heart diseases in a timely manner and assisting clinicians to make more appropriate decisions during labor. However, despite advances in signal processing and machine learning techniques, the analysis of fECG signals has still remained in its preliminary stages. In this work, we describe an algorithm to automatically locate QRS complexes in noninvasive fECG signals obtained from a set of four electrodes placed on the mother’s abdomen. The algorithm is based on an iterative decomposition of the maternal and fetal subspaces and filtering of the maternal ECG (mECG) components from the fECG recordings. Once the maternal components are removed, a novel merging technique is applied to merge the signals and detect the fetal QRS (fQRS) complexes. The algorithm was trained and tested on the fECG datasets provided by the PhysioNet/CinC challenge 2013. The final results indicate that the algorithm is able to detect fetal peaks for a variety of signals with different morphologies and strength levels encountered in clinical practice.


Round Window Membrane Intracochlear Drug Delivery Enhanced by Induced Advection,

David A. Borkholder, XiaoXia Zhu, Robert D. Frisina
Journal of Controlled Release, 174:171-176 (2013).
Publication year: 2013

Delivery of therapeutic compounds to the inner ear via absorption through the round window membrane (RWM) has advantages over direct intracochlear infusions; specifically, minimizing impact upon functional hearing measures. However, previous reports show that significant basal- to-apical concentration gradients occur, with the potential to impact treatment efficacy. Here we present a new approach to inner ear drug delivery with induced advection aiding distribution of compounds throughout the inner ear in the murine cochlea. Polyimide microtubing was placed near the RWM niche through a bullaostomy into the middle ear cavity allowing directed delivery of compounds to the RWM. We hypothesized that a posterior semicircular canalostomy would induce apical flow from the patent cochlear aqueduct to the canalostomy due to influx of cerebral spinal fluid. To test this hypothesis, young adult CBA/CaJ mice were divided into two groups: bullaostomy approach only (BA) and bullaostomy + canalostomy (B+C). Cochlear function was evaluated by distortion product otoacoustic emission (DPOAE) and auditory brainstem response (ABR) thresholds during and after middle ear infusion of salicylate in artificial perilymph (AP), applied near the RWM. The mice recovered for 1 week, and were re-tested. The results demonstrate there was no significant impact on auditory function utilizing the RWM surgical procedure with or without the canalostomy, and DPOAE thresholds were elevated reversibly during the salicylate infusion. Comparing the threshold shifts for both methods, the B+C approach had more of a physiological effect than the BA approach, including at lower frequencies representing more apical cochlear locations. Unlike mouse cochleostomies, there was no deleterious auditory functional impact after 1 week recovery from surgery. The B+C approach had more drug efficacy at lower frequencies, underscoring potential benefits for more precise control of delivery of inner ear therapeutic compounds.


Noninvasive Technique for Monitoring Drug Transport Through the Murine Cochlea using Micro-Computed Tomography

Haghpanahi M, Gladstone MB, Zhu XX, Frisina RD, Borkholder DA
Ann Biomed Eng, 41:2130-2142 (2013) DOI 10.1007/s10439-013-0816-4
Publication year: 2013

Local delivery of drugs to the inner ear has the potential to treat inner ear disorders including permanent hearing loss or deafness. Current mathematical models describing the pharmacokinetics of drug delivery to the inner ear have been based on large rodent studies with invasive measurements of concentration at few locations within the cochlea. Hence, estimates of clearance and diffusion parameters are based on fitting measured data with limited spatial resolution to a model. To overcome these limitations, we developed a noninvasive imaging technique to monitor and characterize drug delivery inside the mouse cochlea using micro-computed tomography (μCT). To increase the measurement accuracy, we performed a subject-atlas image registration to exploit the information readily available in the atlas image of the mouse cochlea and pass segmentation or labeling information from the atlas to our μCT scans. The approach presented here has the potential to quantify concentrations at any point along fluid-filled scalae of the inner ear. This may permit determination of spatially dependent diffusion and clearance parameters for enhanced models.



High-performance, low-voltage electroosmotic pumps with molecularly thin silicon nanomembranes

Jessica L. Snyder, Jirachai Getpreecharsawas, David Z. Fang, Thomas R. Gaborski, Christopher C. Striemer, Philippe M. Fauchet, David A. Borkholder, and James L. McGrath
Proceedings of the National Academies of Sciences, 110(46):18425-18430 (2013)
Publication year: 2013

Electroosmotic pumps (EOPs) are a class of pumps in which fluid is driven through a capillary or porous media within an electric field. Current research on EOPs concerns the development of new materials in which high electroosmotic flow rates can be achieved for low voltages. Such pumps could be used for portable microfluidic devices. Porous nanocrystalline silicon (pnc-Si) is a material that is formed into a 15-nm-thick nanomembrane. pnc-Si membranes are shown here to have high electroosmotic flow rates at low applied voltages due to the high electric fields achieved over the ultrathin membrane. A prototype EOP was designed using pnc-Si membranes and shown to pressurize fluid through capillary tubing at voltages as low as 250 mV.


Microsystems technologies for drug delivery to the inner ear

Leary Pararas EE, Borkholder DA, Borenstein JT
Advanced Drug Delivery Reviews, Advanced Drug Delivery Reviews, 64(14):1650-1660, (2012)
Publication year: 2012

The inner ear represents one of the most technologically challenging targets for local drug delivery, but its clinical significance is rapidly increasing. The prevalence of sensorineural hearing loss and other auditory dis- eases, along with balance disorders and tinnitus, has spurred broad efforts to develop therapeutic compounds and regenerative approaches to treat these conditions, necessitating advances in systems capable of targeted and sustained drug delivery. The delicate nature of hearing structures combined with the relative inaccessibility of the cochlea by means of conventional delivery routes together necessitate significant advancements in both the precision and miniaturization of delivery systems, and the nature of the molecular and cellular targets for these therapies suggests that multiple compounds may need to be delivered in a time- sequenced fashion over an extended duration. Here we address the various approaches being developed for inner ear drug delivery, including micropump-based devices, reciprocating systems, and cochlear prosthesis-mediated delivery, concluding with an analysis of emerging challenges and opportunities for the first generation of technologies suitable for human clinical use. These developments represent exciting advances that have the potential to repair and regenerate hearing structures in millions of patients for whom no currently available medical treatments exist, a situation that requires them to function with electronic hearing augmentation devices or to live with severely impaired auditory function. These advances also have the potential for broader clinical applications that share similar requirements and challenges with the inner ear, such as drug delivery to the central nervous system.


In-Plane Biocompatible Microfluidic Interconnects for Implantable Microsystems

Johnson, D., Frisina, R., Borkholder, D.A
IEEE Transactions on Biomedical Engineering, 58(4):943-948 (2011)
Publication year: 2011

Small mammals, particularly mice, are very useful animal models for biomedical research. But extremely small anatomical dimensions make design of implantable microsystems quite challenging. A method for coupling external fluidic systems to microfluidic channels via in-plane interconnects is presented. Capillary tubing is inserted into channels etched in the surface of a silicon wafer with a seal created by Parylene-C deposition. A model has been developed based on Parylene-C deposition characterizations which allows prediction of deposition into tapered channels for design optimization. Low volume interconnects using biocompatible, chemical resistant materials have been demonstrated and shown to withstand pressure as high as 827 kPa (120 psi) with average pull test strength of 2.9 N. Each interconnect consumes less than 0.018 mm3 (18 nl) of volume. The low added volume makes this an ideal interconnect technology for medical applications where implant volume is critical.


Murine intracochlear drug delivery: Reducing concentration gradients within the cochlea

Borkholder, D.A., Xiaoxia Zhu, Brad T. Hyatt, Alfredo S. Archilla, William J. Livingston III, and Robert D. Frisina
Hearing Research, 268(1-2):2-11 (2010)
Publication year: 2010

Direct delivery of compounds to the mammalian inner ear is most commonly achieved by absorption or direct injection through the round window membrane (RWM), or infusion through a basal turn coch- leostomy. These methods provide direct access to cochlear structures, but with a strong basal-to-apical concentration gradient consistent with a diffusion-driven distribution. This gradient limits the efficacy of therapeutic approaches for apical structures, and puts constraints on practical therapeutic dose ranges. A surgical approach involving both a basal turn cochleostomy and a posterior semicircular canal canal- ostomy provides opportunities for facilitated perfusion of cochlear structures to reduce concentration gradients. Infusion of fixed volumes of artificial perilymph (AP) and sodium salicylate were used to evaluate two surgical approaches in the mouse: cochleostomy-only (CO), or cochleostomy-plus-canal- ostomy (CþC). Cochlear function was evaluated via closed-system distortion product otoacoustic emis- sions (DPOAE) threshold level measurements from 8 to 49 kHz. AP infusion confirmed no surgical impact to auditory function, while shifts in DPOAE thresholds were measured during infusion of salicylate and AP (washout). Frequency dependent shifts were compared for the CO and CþC approaches. Computer simulations modeling diffusion, volume flow, interscala transport, and clearance mechanisms provided estimates of drug concentration as a function of cochlear position. Simulated concentration profiles were compared to frequency-dependent shifts in measured auditory responses using a cochlear tonotopic map. The impact of flow rate on frequency dependent DPOAE threshold shifts was also evaluated for both surgical approaches. Both the CþC approach and a flow rate increase were found to provide enhanced response for lower frequencies, with evidence suggesting the CþC approach reduces concentration gradients within the cochlea.


State-of-the-art mechanisms of intracochlear drug delivery

Borkholder, D.A
Current Opinions in Otolaryngology & Head and Neck Surgery, 16:472-477 (2008).
Publication year: 2008

Purpose of review

Treatment of auditory and vestibular dysfunction has become increasingly dependent on inner ear drug delivery. Recent advances in molecular therapy and nanotechnology have pushed development of alternate delivery methodologies involving both transtympanic and direct intracochlear infusions. This review examines recent developments in the field relevant to both clinical and animal research environments.

Recent findings

Transtympanic delivery of gentamicin and corticosteroids for the treatment of Meniere’s disease and sudden sensorineural hearing loss continues to be clinically relevant, with understanding of pharmacokinetics becoming more closely studied. Stabilizing matrices placed on the round window membrane for sustained passive delivery of compounds offer more controlled dosing profiles than transtympanic injections. Nanoparticles are capable of traversing the round window membrane and cochlear membranous partitions, and may become useful gene delivery platforms. Cochlear and vestibular hair cell regeneration has been demonstrated by vector delivery to the inner ear, offering promise for future advanced therapies.


Optimal methods of inner ear drug delivery will depend on toxicity, therapeutic dose range, and characteristics of the agent to be delivered. Advanced therapy development will likely require direct intracochlear delivery with detailed understanding of associated pharmacokinetics.

Microchip Isoelectric Focusing Using a Miniature Scanning Detection System

Raisi, F., Belgrader, P., Borkholder, D. A., Herr, A. E., Kintz, G. J., Pourhamadi, F., Taylor, M. T., and Northrup, M. A.
Electrophoresis, 22(11):2291-2295 (2001)
Publication year: 2001

A miniature scanning fluorescent detector has been developed for plastic microchannel isoelectric focusing (mIEF) analysis. The detector, comprised of a lamp and photomultiplier tube (PMT) on a moving stage, measured the real-time distribution of fluorescently labeled peptides subjected to gel-free mIEF. During the run, the effective length of the 6-cm channel was scanned every 9 s. Analysis was completed within 5 min while still obtaining high resolution and sensitivity. In addition, the scanning detector was used to characterize peptide migration properties within the channel by providing simultaneous temporal and spatial measurements.

A Microfluidic Cartridge to Prepare Spores for PCR Analysis

Belgrader, P., Okuzumi, M., Pourahmadi, F., Borkholder, D.A., and Northrup, M.A
Biosensors & Bioelectronics 14:849-852 (2000)
Publication year: 2000

A prototype cartridge system is described that rapidly disrupts Bacillus spores by sonication, adds PCR reagent to the disrupted spores, and dispenses the mixture into a PCR tube. The total time to automatically process the spores in the cartridge and then detect the spore DNA by real-time PCR was 20 min.


Development and Application of Cell-Based Biosensors

Pancrazio, J.J., Whelan, J. P., Borkholder, D.A., Ma, W. and Stenger, D.A.
Annals of Biomedical Engineering, 27:697–711 (1999).
Publication year: 1999

Biosensors incorporate a biological sensing element that converts a change in an immediate environment to signals conducive for processing. Biosensors have been implemented for a number of applications ranging from environmental pollutant detection to defense monitoring. Biosensors have two intriguing characteristics: (1) they have a naturally evolved selectivity to biological or biologically active analytes; and (2) biosensors have the capacity to respond to analytes in a physiologically relevant manner. In this paper, molecular biosensors, based on antibodies, enzymes, ion channels, or nucleic acids, are briefly reviewed. Moreover, cell-based biosensors are reviewed and discussed. Cell-based biosensors have been implemented using microorganisms, particularly for environmental monitoring of pollutants. Biosensors incorporating mammalian cells have a distinct advantage of responding in a manner that can offer insight into the physiological effect of an analyte. Several approaches for transduction of cellular signals are discussed; these approaches include measures of cell metabolism, impedance, intracellular potentials, and extracellular potentials. Among these approaches, networks of excitable cells cultured on microelectrode arrays are uniquely poised to provide rapid, functional classification of an analyte and ultimately constitute a potentially effective cell-based biosensor technology. Three challenges that constitute barriers to increased cell-based biosensor applications are presented: analytical methods, reproducibility, and cell sources. Possible future solutions to these challenges are discussed.


Portable Cell-Based Biosensor System for Toxin Detection

Pancrazio, J.J., Bey, P.P. Jr., Cuttino, D.S., Kusel, J.K., Borkholder, D.A.
Sensors and Actuators B: Chemical, 53(3):179-185 (1998)
Publication year: 1998

A portable cell-based biosensor has been developed and characterized. The prototype system relies on extracellular recording from excitable cells cultured over an array of platinized gold microelectrodes. Extracellular potentials were bandpass filtered between 80 Hz to 2.8 kHz and amplified with a selectable gain of either 1000 or 5000. The input-referred noise level of the system was only 8.7 μVRMS in the laboratory setting, reaching only 10.6 μVRMS in an outdoor environment, more than sufficient for measurement of extracellular potentials from excitable cells. The system also incorporates a feedback control system for temperature regulation and a 36-channel multiplexer for selection of up to four output channels for simultaneous display. Wherever possible, low-cost ‘off-the-shelf’ components were utilized in this prototype biosensor design. Using this system, extracellular recordings from chick myocardiocytes were performed under both laboratory and outdoor conditions.



Description and Demonstration of a CMOS Amplifier-Based-System with Measurement and Stimulation Capability for Bioelectrical Signal Transduction

Pancrazio J.J., Bey, P.P.Jr., Loloee, A., Manne, S., Chao, H.C., Howard, L.L., Gosney, W.M., Borkholder, D.A., Kovacs, G.T.A., Manos, P., Cuttino, D.S. and Stenger, D.A.
Biosensors and Bioelectronics 13:971-979 (1998).
Publication year: 1998

An extracellular recording system incorporating an electrode array and an amplifier/stimulator CMOS chip is described and characterized. Important features of this custom VLSI chip include 16 instrumentation amplifiers with a gain of 50 and the incorporation of a cross-point array allowing designation of an extracellular microelectrode as either a stimulator or sensor. The planar array consisted of 32 microelectrodes, 14 mm in diameter, and four larger reference electrodes. Microelectrodes, interconnecting traces, and bond pads were patterned with a 500-nm layer of gold. The interconnecting traces were passivated with a 1-mm thick layer of silicon nitride to provide chemical and electrical insulation and microelectrode impedance was lowered utilizing electrode position of platinum black. The amplifier exhibited a nearly flat frequency response with high pass and low pass corner frequencies of 0.7 Hz and 50 kHz, respectively. The input referred noise over the 50 kHz bandwidth was 12–16 mVRMS, well below the magnitude of previously reported extracellular potentials. Crosstalk between neighboring channels resulted in an output signal below the amplifier noise level, even for relatively large extracellular potentials. Using this system, extracellular recordings were demonstrated yielding typical peak-to-peak biopotentials of magnitude 0.9–2.1 mV and 100–400 mV for chick cardiac myocytes and rat spinal cord neurons, respectively. The key components of this extracellular recording system can be manufactured using industry standard thin film photolithographic techniques.


Cell-based Sensor Microelectrode Array Characterized by Imaging X-ray Photoelectron Spectroscopy, Scanning Electron Microscopy, Impedance Measurements, and Extracellular Recordings

Jung, D.R., Cuttino, D.S., Pancrazio, J.J., Manos, P., Cluster, T., Sathanoori, R.S., Aloi, L.E., Coulombe, M.G., Bey, P., Czarnaski, M.A., Borkholder, D.A., Kovacs, G.T.A., Stenger, D.A. and Hickman, J.J.
Journal of Vacuum Science & Technology A, 16(3):1183-1188 (1998).
Publication year: 1998

We are developing a cell-based biosensor consisting of a planar microelectrode array that allows detection of extracellular potentials and their modulation in the presence of toxins or other active agents. To improve cell–electrode coupling, the microelectrodes were electroplated with platinum black. We report on the use of imagingx-ray photoelectron spectroscopy(XPS),scanning electron microscopy(SEM), impedance measurements, and extracellular recordings to assess the effectiveness of this procedure. SEM provided highly detailed images of the shape and structure of well-formed deposits of thickness on the order of 1 μm or more. Because of its inherent high surface sensitivity, imagingXPS could reveal the presence of platinum deposits that were too thin to be detected by SEM. For typical, well-plated microelectrodes, impedance measurements showed reductions in the electrical resistance at 100 Hz from roughly 60 MΩ or more 1 MΩ. The overall electronic coupling of biopotentials to the microelectrodes was demonstrated by recordings obtained from beating rat myocytes and from rat spinal cord cells.


Plasma-Etched Neural Probes

Kewley, D.T., Hills, M.D., Borkholder, D.A., Opris, I.E., Maluf, N.I., Storment, C.W., Bower, J.M. and Kovacs, G.T.A.
Sensors and Actuators: A Physical, 58:27-35, (1997)
Publication year: 1997

A new method is presented for microfabricating silicon-based neural probes that are designed for neurobiology research. Such probes provide unique capabilities to record high-resolution signals simultaneously from multiple, precisely defined locations within neural tissue. The fabrication process utilizes a plasma etch to define the probe outline, resulting in sharp tips and compatibility with standard CMOS processes. A low-noise amplifier array has been fabricated through the MOSIS service to complete a system that has been used in multiple successful physiological experiments.



Microelectrode Arrays for Stimulation of Neural Slice Preparations

Borkholder, D., Bao, J., Maluf, N., Perl, E., and Kovacs, G.
Journal of Neuroscience Methods, 77:61-66 (1997)
Publication year: 1997

A planar 6×6 array of iridium electrodes with four reference electrodes has been developed for use with neural tissue preparations. Precise knowledge of the relative locations of the array elements allows for spatial neurophysiological analyses. The 10 μm diameter platinized iridium electrodes on a 100 μm pitch have been used to stimulate acutely prepared slices of spinal cord from free-ranging rodents. An intracellular recording from a single neuron in the substantia gelatinosa (SG) using the whole-cell, tight-seal technique allowed low noise, high resolution studies of excitatory or inhibitory electrical responses of a given neuron to inputs from the primary afferent fibers or from stimulation by individual electrodes of the array. The resulting maps of responses provide an indication of the interconnectivity of neural processes. The pattern emerging is that of limited interconnectivity in the SG from areas surrounding a recorded neuron but with strong excitatory or inhibitory effects from those oriented in a longitudinal (rostral–caudal) direction relative to the neuron. The observations to date suggest the neurons of the SG are arranged in sets of independent networks, possibly related to sensory modality and input from particular body regions.



Quarter-Micron Lithography with a Gapped Markle-Dyson System

Owen, G., Borkholder, D.A., Knorr, C., Markle, D.A. and Pease, R.F.W.
Journal of Vacuum Science & Technology B, 12(6):3809-3813 (1994)
Publication year: 1994

0.25‐μm lithography has previously been demonstrated using an ungapped prototype Markle–Dyson system, in which the wafer was held in soft contact with the mask. However, such an in‐contact scheme would be inappropriate for semiconductor fabrication, and so a second prototype has been designed and constructed, in which a gap of 25 μm is introduced between the mask and the wafer. The two major technical problems to be overcome in implementing the gap are the measurement and setting of the gap itself, and correction for the spherical aberration which it introduces. In the new prototype, the gap is set by a piezo‐electric actuator and measured using a capacitance gauge, and the spherical aberration is corrected by using a mirror which deviates slightly (by 0.33 μm) from sphericity. The system has been tested lithographically, using chromium reflective masks. It has demonstrated a resolution of 0.25 μm in Shipley XP89131photoresist.

Flexible, Dry-Released Process for Aluminum Electrostatic Actuators

Storment, C.W., Borkholder, D.A., Westerlind, V., Suh, J.W., Maluf, N.I. and Kovacs, G.T.A.
IEEE J. Microelectromechanical Systems, 3(3):90-6 (1994)
Publication year: 1994

We present an all-aluminum MEMS process (Al-MEMS) for the fabrication of large-gap electrostatic actuators with process steps that are compatible with the future use of underlying, pre-fabricated CMOS control circuitry. The process is purely additive above the substrate as opposed to processes that depend on etching pits into the silicon, and thereby permits a high degree of design freedom. Multilayer aluminum metallization is used with organic sacrificial layers to build up the actuator structures. Oxygen-based dry etching is used to remove the sacrificial layers. While this approach has been previously used by other investigators to fabricate optical modulators and displays, the specific process presented herein has been optimized for driving mechanical actuators with relatively large travels. The process is also intended to provide flexibility for design and future enhancements. For example, the gap height between the actuator and the underlying electrode(s) can be set using an adjustable polyimide sacrificial layer and aluminum “post” deposition step. Several Al-MEMS electrostatic structures designed for use as mechanical actuators are presented as well as some measured actuation characteristics