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Abstract Biosensors are analytical tools used for the analysis of biomaterial samples and provide an understanding about the biocomposition, structure, and function of biomolecules and/or biomechanisms by converting the biological response into an electrical and/or optical signal. In particular, with the rise in antibiotic resistance amongst pathogenic bacteria, the study of antibiotic activity and transport across cell membranes in the field of biosensors has been gaining widespread importance. Herein, for the rapid and label‐free detection of antibiotic permeation across a membrane, a microelectrode integrated microfluidic device is presented. The integrated chip consists of polydimethylsiloxane based microfluidic channels bonded onto microelectrodes on‐glass and enables us to recognize the antibiotic permeation across a membrane into the model membranes based on electrical impedance measurement, while also allowing optical monitoring. Impedance testing is label free and therefore allows the detection of both fluorescent and non‐fluorescent antibiotics. As a model membrane, Giant Unilamellar Vesicles (GUVs) are used and impedance measurements were performed by a precision inductance, capacitance, and resistance metre. The measured signal recorded from the device was used to determine the change in concentration inside and outside of the GUVs. We have found that permeation of antibiotic molecules can be easily monitored over time using the proposed integrated device. The results also show a clear difference between bilayer permeation that occurs through the lipidic bilayer and porin‐mediated permeation through the porin channels inserted in the lipid bilayer.

High-frequency irreversible electroporation (H-FIRE) is an emerging cancer therapy, which uses bursts of short duration, alternating polarity, high-voltage electrical pulses to focally ablate tumors. Here, we present a preliminary investigation of the combinatorial effects of H-FIRE and ionizing radiation. In vitro cell cultures were exposed to bursts of 500 ns pulses and single radiation doses of 2 or 20 Gy then analyzed for 14 days. H-FIRE and radiation therapy (RT) appear to induce different delayed cell death mechanisms and in all treatment groups combinatorial therapy resulted in lower overall viabilities. These results indicate that in vivo investigation of the antitumor efficacy of combined H-FIRE and RT is warranted. [ABSTRACT FROM PUBLISHER]

Dielectric spectroscopy (DS) is a non‐invasive, label‐free, and promising technique for measuring dielectric properties of biological cells. Recent developments in microfabrication techniques made it possible to perform DS measurements with minute volume of cell suspensions. However, when the cell size approaches the size of the measurement chamber, especially, for single cell measurements, the analytical models [Maxwell–Wagner and Bruggeman–Hanai (BH) mixture models] to extract cell parameters lose their accuracy. Moreover, variations in the cell position relative to the measurement electrodes decrease the accuracy of the analytical solutions. Impedance spectrum of a typical eukaryotic mammalian cell is generated for different geometrical configurations using finite element. The generated data are fitted to the analytical models and extracted cell parameters are compared with the original values. The results show that BH model works more effectively when chamber to cell radius ratio is <3.5 and chamber height to cell radius ratio is <3. Moreover, it is observed that analytical models estimate cell parameters with major errors when the cells are in the vicinity of the electrodes. However, for high‐volume fraction simulations, the BH model was able to predict cell parameters better even in the vicinity of the electrodes. [ABSTRACT FROM AUTHOR]

Purpose: The objective of this study is to measure through simulation the impact of (1) heterogeneity of biophysical parameters in tumor vs healthy tissue, (2) applicator placement relative to the tumor, and (3) proximity to large blood vessels on microwave ablation (MWA) treatment effect area. This will help identify the biophysical properties that have the greatest impact on improving clinical modeling of MWA procedures. Methods: The authors' approach was to develop two-compartment models with variable tissue properties and simulate MWA procedures performed in liver with Perseon Medical's 915 MHz short-tip applicator. Input parameters for the dielectric and thermal properties considered in this study were based on measurements for healthy and malignant (primary or metastatic) liver? tissue previously reported in the literature. Compartment 1 (C1) represented normal, fatty, or cirrhotic liver, and compartment 2 (C2) represented a primary hepatocellular carcinomatumor sample embedded within C1. To evaluate the sensitivity to tissue parameters, a range of clinically relevant tissue properties were simulated. To evaluate the impact of MWA antenna position, the authors simulated various tumor perfusion models with the antenna shifted 5 mm anteriorly and posteriorly. To evaluate the effect of local vasculature, the authors simulated an additional heat sink of various diameters and distances from the tumor. Dice coefficient statistics were used to evaluate ablation zone effects from these local heat sinks. Results: Models showed less than 11% of volume variability (1 cm³ increase) in ablation treatment effect region when accounting for the difference in relative permittivity and electrical conductivity between malignant and healthy liver? tissue. There was a 27% increase in volume when simulating thermal conductivity of fatty liver disease versus the baseline simulation. The ablation zone volume increased more than 36% when simulating cirrhotic surrounding liver? tissue. Antenna placement relative to the tumor had minimal sensitivity to the absolute size of the treatment effect area, with less than 1.5 mm variation. However, when considering the overlap between the ablation zone and the ideal clinical margin when the antenna was displaced 5 mm anteriorly and posteriorly, there was approximately a 6 mm difference in the margins. Dice coefficient statistics showed as much as an 11% decrease in the ablation margin due to the presence of vessel heat sinks within the model. Conclusions: The results from simulating the variance in malignant tissue thermal and electrical properties will help guide better approximations for MWA treatments. The results suggest that assuming malignant and healthy liver tissues have similar dielectric properties is a reasonable first approximation. Antenna placement relative to the tumor has minimal impact on the absolute size of the ablation zone, yet it does cause relevant variation between desired treatment margin and ablation zone. Blood vessel cooling, especially hepatic vessels close to the region of interest, may be a significant factor to consider in treatment planning. Further data need to be collected for assessing treatment planning utility of modeling MWA in this context. [ABSTRACT FROM AUTHOR]

Microstencils have been utilized in biomedical engineering to pattern cell cultures, engineer tissues, and pattern conductive materials in microelectronics for the measurement of bioelectric activity. However, fabricating these microstencils can be considerably time consuming, expensive, and cleanroom processing or laser micromachining intensive. We present microfabrication strategies for producing stencils rapidly and cost effectively, with minimal use of cleanroom facilities, ideal for prototyping or applications where microfabrication costs need to be conserved. The process utilizes 3-D-printed templates as master structures from, which polydimethylsiloxane (PDMS) microstencils are molded. The entire process, from concept to completed stencil, requires approximately one day; however, the majority of this time is budgeted for PDMS curing cycles, so it requires only ~1-2 person hours to complete. These microstencils were used to pattern metal traces ~160-1000-μm wide, on three commonly used BioMEMS substrate materials- to pattern rat cortical neuronal cell cultures with radii between ~300 and 1000 μm-and to pattern organic materials. With the advancement of 3-D-printing technologies, we anticipate that our presented processes will improve in resolution and gain a greater cost advantage over traditional microstencilling methods. [ABSTRACT FROM AUTHOR]

When extracellular action potentials (spikes) from cultured neurons are recorded using microelectrode arrays in open wells, their amplitudes are usually quite small (often below the noise level) despite the extracellular currents originating from the relatively large surface area of neural cell somata. In this paper rat cortical neurons were seeded into one well of a two well system separated by 3 ×10 μm microtunnels and then seven days later into the second well forming a feed-forward network between two small neuronal assemblies. In contrast to measurements in the open well spikes recorded from axons within the restricted volumes imposed by the microtunnels are often several orders of magnitude larger than in the open well, with high signal to noise ratio, despite the currents originating in the much smaller surface area of the axon. Average signal amplitudes exceeding 250 μV are typical, with some signals as large as 4.5 mV (signal-to-noise ratio up to 450), 20 times greater than the maximum recorded from electrodes in adjacent but open wells. We confirm the dependence of signal amplitude on the impedance of the microtunnel and discuss possible reasons for the phenomenon. [ABSTRACT FROM AUTHOR]

A novel microdevice for electrical impedance spectroscopy is presented to differentiate between normal and cancerous cells at the single cell level with a high sensitivity. The device utilizes a microfluidic tunnel structure, whose height is smaller than the cell diameters, to squeeze the target cells. Thus, the tight contact between the cells and electrodes allows the device to measure the intrinsic electrical properties of the cells with a higher sensitivity than through noncontact methods. Three-dimensional interdigitated electrodes (3-D IDE) are also employed to confine most of the electric field into the cell membrane, which contains the meaningful electrical properties. Consequently, the variance of the measured values could be reduced because the position of target cells is almost the same in every cell assay. The device well distinguishes normal human breast cells (MCF-10A) from early-stage human breast cancer cells (MCF-7) by means of electrical impedances at 500 kHz; the average differences of the real part and phase angle between the target cells are 44.4 and 1.41 , respectively. The proposed device also shows a high repeatability for the deionized (DI) water test conducted before and after each cell assay; the variances of the real part and phase angle of electrical impedance at 500 kHz are as small as 9.07 and 0.27 , respectively. [ABSTRACT FROM PUBLISHER]

Presented is a noncontact model and radiometric sensor developed to facilitate core body temperature extraction. The system has been designed as a close-proximity sensor to detect thermal emissions radiated from deep inside the human body. The radiometer uses a cavity-backed slot antenna (CBSA) designed to account for performance degradation which occurs in the near field of the human body. Tissue-simulating materials with electrical properties similar to the human body have been identified, and layered configurations of these phantoms are used to mimic the electromagnetic characteristics of a human stomach volume: hence, defines the core model. Positioned approximately 7 mm from the core model, the sensor tracks the change in brightness temperature as the subsurface physical temperature is varied. A mathematical noncontact model (NCM) is subsequently used to correlate the observed brightness temperature to the subsurface temperature, accounting for artifacts induced by the sensor's remote positioning from the specimen. The accuracy of the NCM is validated through an analysis of the measurement data extracted from the test bed before and after applying the model. The results illustrate that the measurement is highly sensitive to the antenna impedance match and atmospheric temperature. The measurement is less sensitive to the physical temperature of the instrument and emissivity of the specimen. The results of this study demonstrate that radiometric sensors are capable of close proximity, subsurface extraction of biological data given that certain parameters are closely monitored. [ABSTRACT FROM PUBLISHER]

A biocompatible neural microprobe constructed using well-established SU-8 microfabrication techniques is described that was designed to record fiber spike signals from regenerated axons within peripheral nerves. These microprobes features bipolar longitudinal gold electrodes recessed below the surface within ldquogroovesrdquo designed to guide the growth of regenerating axons along the length of the grooves and limit the number of fibers that come in contact with the longitudinal electrodes. In addition, screening microprobe toxicity using cultures of human skin fibroblasts, the biocompatibility of these SU-8 microprobes for neural interface applications, in particular, was specifically verified using primary cultures of two sensitive cell types found in peripheral nerves: purified Schwann cells and explanted dorsal root ganglion (DRG) neurons and their fibers. The SU-8 microprobes were surgically implanted into transected rat Sciatic nerves within a unique peripheral nerve regeneration tube. Long-term fiber spike signals were recorded with these SU-8 microprobes in 13 chronically implanted rats for periods from 4 to 51 weeks without any signs of tissue damage or inflammatory reaction. [ABSTRACT FROM PUBLISHER]

A platform for high spatial and temporal resolution electrophysiological recordings of in vitro electrogenic cell cultures handling 4096 electrodes at a full frame rate of 8 kHz is presented and validated by means of cardiomyocyte cultures. Based on an active pixel sensor device implementing an array of metallic electrodes, the system provides acquisitions at spatial resolutions of 42 mum on an active area of 2.67 mm times 2.67 mm, and in the zooming mode, temporal resolutions down to 8 mus on 64 randomly selected electrodes. The low-noise performances of the integrated amplifier (11 muVrms) combined with a hardware implementation inspired by image/video processing concepts enable high-resolution acquisitions with real-time preprocessing capabilities adapted to the handling of the large amount of acquired data. [ABSTRACT FROM PUBLISHER]

We present a system for the online, in vitro, nondestructive monitoring of tissue growth within microporous polymer scaffolds. The system is based on measuring the admittance of the sample over a frequency range of 10-200 MHz using an open-ended coaxial probe and impedance analyzer. The sample admittance is related to the sample complex permittivity (CP) by a quasi-static model of the probe's aperture admittance. A modified effective medium approximation is then used to relate the CP to the cell volume fraction. The change of cell volume fraction is used as a measure of tissue growth inside the scaffold. The system detected relative cell concentration differences between microporous polymer scaffolds seeded with 0.4, 0.45, 0.5, and 0.6 x 106 pre-osteoblast cells. In addition, the pre-osteoblast proliferation within 56 scaffolds over 14 days was recorded by the system and a concurrent DNA assay. Both techniques produced cell proliferation curves that corresponded to those found in literature. Thus, our data confirmed that the new system can assess relative cell concentration differences in microporous scaffolds enabling online nondestructive tissue growth monitoring. [ABSTRACT FROM PUBLISHER]

We describe an integrated circuit for sensing the substrate coupling capacitance of anchorage-dependent living cells in a standard culture environment. Capacitance is measured using charge redistribution in response to weak, low frequency electric field excitations. The underlying biophysical phenomenon results primarily from the insulating nature of the cell structure and the counterionic polarization in the surrounding aqueous medium. The measured capacitance depends on a variety of factors related to the cell, its growth environment and the supporting substrate. These include membrane integrity, morphology, extracellular ionic concentration, adhesion strength, and substrate proximity. The measured capacitance is an indication of both the interaction between cells and substrate and cell health. The capacitance sensor uses the principle of charge sharing and translates sensed capacitance values to output voltages. The sensor chip has been fabricated in a commercially available 0.5-mum, 2-poly 3-metal CMOS technology. The sensing technique does not require direct electrical connection to the aqueous culture medium. We report results from experiments demonstrating on-chip tracking of cell adhesion as well as long term monitoring of cell viability in vitro [ABSTRACT FROM PUBLISHER]

We describe the use of compact green light-emitting diodes (LEDs) for fluorescence optical imaging and demonstrate this for dynamic recording from cultured neurons. The efficient gallium-nitride-based LEDs have individual element sizes comparable to typical biological cells and are operated in proximity illumination mode for individual neurons. As periodic arrays, and with direct electrical control of each LED, a spatially periodic multicellular target can be induced to fluoresce in a predesigned spatiotemporal sequence, thereby providing a new approach to dynamic recording of small neural circuits. [ABSTRACT FROM PUBLISHER]

In in vitro tissue engineering, microporous scaffolds are commonly used to promote cell proliferation and differentiation in three-dimensional structures. Classic measurement methods are particularly time consuming, difficult to handle, and destructive. In this study, a new nondestructive method based on complex permittivity measurement (CPM) is proposed to monitor and track the osteoblast and macrophage differentiation through their morphological variation upon cell attachment and proliferation inside the microporous scaffolds. CPM is performed using a vector network analyzer and a dielectric probe under sterile conditions in a laminar-flow hood. A suitable effective medium approximation (EMA) is applied to fit the data in order to extract the parameters of the different constituents. Our data show that the EMA depolarization factor can be monitored to assess the variation of cell morphology characterizing cell attachment. Discrimination between two batches of scaffolds seeded, respectively, with 2 million and 1 million osteoblast cells is possible; the ratio of their CPM-derived cell volume fractions is in agreement with the ratio of their cell seeding numbers. In addition, cell proliferation inside scaffolds seeded with osteoblasts cultured in alpha minimum essential medium and inside scaffolds seeded with osteoblasts cultured in alpha minimum essential medium supplemented to induce the formation of extracellular matrix is monitored via CPM over several days. CPM-determined cell volume fraction is compared to DNA assay cell counts. Extracellular matrix formation and cell presence was confirmed by scanning electron microscopy. A set of three signature parameters (ε'mem,ε'cyt,κ'cyt) characteristic of cell line is extracted from CPM. Distinct signatures are recorded for osteoblasts and macrophages, thus confirming the ability of CPM to discriminate between different cell types. This study demonstrates the potential of CPM as a diagnostic tool to monitor quickly and noninvasively cell growth and differentiation inside microporous scaffolds. Our findings suggest that the use of CPM could be extended to many biomedical applications, such as drug detection and automation of tissue and bacterial cultures i- n bioreactors. [ABSTRACT FROM PUBLISHER]

Studies of electrically induced morphological changes in neurons have either been limited by the resolution of light microscopy or the cell fixation required for electron microscopy. Atomic force microscopy (AFM), however, mechanically maps cell topography, offering exquisite resolution of evolving processes in three dimensions. In this paper, we present a microelectrode array (MEA) based platform for the real-time detection of subtle, electrically induced variations in neuronal morphology, with AFM. This platform required the customized design and production of a silicon-based MEA, integration with a commercial AFM, and the development of biological techniques for culture of neuroblastoma (SH-SY5Y) cells onto the device. Biphasic pulse trains (1 Hz) of electric current were delivered to a microelectrode interfaced with a neuroblastoma cell, and the AFM continuously recorded a cross-sectional height profile. Proof-of-principle experiments demonstrate that electric stimulation may induce fluctuations ranging in the 100-300-nm range, 75-fold greater than the systemic resolution, but smaller than the resolution of light microscopy modalities. In addition, the real-time capabilities of AFM captured a collapse (30%-40%) of a neurite cross section, seconds after electric stimulation. Ultimately, this platform can be used to nanocharacterize cell responses to electric stimulation and other biochemical cues, for use in neuronal patterning and regeneration studies. [ABSTRACT FROM PUBLISHER]

Presents a lab-on-a-chip for electronic manipulation and detection of microorganisms based on the use of closed dielectrophoretic (DEP) cages combined with impedance sensing. A printed circuit board (PCB) prototype has been used to trap, concentrate, and quantify polystyrene micro-beads in agreement with CAD simulations. The experiment was successfully repeated with S. cerevisiae. The results prove the effectiveness of the approach for particle manipulation and detection without the need for external optical components nor chemical labeling. With the proposed approach, particle concentration may be increased on-chip of more than three orders of magnitude, correspondingly boosting the detection sensitivity. [ABSTRACT FROM PUBLISHER]

This paper describes the adhesion and growth of dissociated cortical neurons on chemically patterned surfaces over a time period of 30 days. The presence of neurons was demonstrated by measurement of spontaneous bioelectrical activity on a micropatterned multielectrode array. Chemical patterns were prepared with a combination of neurophobic layers of polyethylenoxide-polypropylenoxide-polyethylenoxide (PEO-PPO-PEO) triblockcopolymers adsorbed onto hydrophobic surfaces and neurophilic microprinted tracks of polyethylenimine (PEI). Results showed that commercially available PEO-PPO-PEO triblockcopolymers F108 and F127 (Synperonics, ICI) significantly reduced the adhesion of neuronal tissue when adsorbed on hydrophobic Polyimide (PI) and Fluorocarbon (FC) surfaces over a time period of eight days. In general, both F108- and F127-coated PI displayed equal or better neurophobic background properties after 30 days. Viability of neuronal tissue after 30 days on PEI microprinted F108- and F127-coated PI was comparable with relatively high viability factors between 0.9 and 1 (scale from 0 to 1). Summarizing, the strategy to combine the neurophobic adsorbed triblock-copolymers F108 and F127 onto hydrophobic surfaces with neurophilic microprinted PEI resulted in relatively long-term neuronal pattern preservation with high numbers of viable neurons present after 30 days. [ABSTRACT FROM PUBLISHER]

The magnetic orientations of human red blood cells (RBCs) in some media such as phosphate buffer solutions (PBS), serum and plasma were investigated to ascertain the behavior of RBCs under strong magnetic fields in the body. The effects of orientation of RBCs in suspensions were measured by changes in the electrical conductivity of suspensions. The electrical conductivity of the suspensions were increased by application of magnetic fields parallel to the electric field. This phenomenon indicates that RBCs are redirected by a magnetic field. The changes in electrical conductivity of samples of 40% RBCs suspended in PBS, serum and plasma were 19%, 9% and less than 4% respectively. The electrical conductivity of anticoagulated whole blood was not changed by application of magnetic fields up to 8 T. Through microscopic observation of RBCs suspended in plasma under 8 T magnetic field, it was revealed that the aggregation of RBCs or rouleau formation occurred in plasma and rouleaux were oriented by the strong magnetic field. The changes in the electrical conductivity of the suspensions and whole blood by the magnetic orientation are prevented by the aggregation and rouleaux formation of RBCs. [ABSTRACT FROM PUBLISHER]

Presented is a methodology of conversion of a conventional colour measure, e.g. 24‐bit colour, to a neuromorphic representation for use with epiretinal prostheses. The conversion implemented, for a 4 by 4 image, within a field programmable gate array outputs three groups of timing signals. These three groups correspond to red (R), green (G) and blue (B) colour channels. In future work the channels will input to a retinal implant stimulator, via a receiver chip housed over the earlobe of a prosthesis recipient, to form biphasic pulses to drive electrodes stimulating the axons of colour specific retinal ganglion cells. A test setup was used to encode the hue pulses back to intensity to display test images on a VGA monitor for validation. [ABSTRACT FROM AUTHOR]

Nonthermal irreversible electroporation (NTIRE) is an emerging tissue ablation modality that may be ideally suited in developing a decellularized tissue graft. NTIRE utilizes short electric pulses that produce nanoscale defects in the cell membrane lipid bilayer. The electric parameters can be chosen in such a way that Joule heating to the tissue is minimized and cell death occurs solely due to loss in cell homeostasis. By coupling NTIRE with the body's response, the cells can be selectively ablated and removed, leaving behind a tissue scaffold. Here, we introduce two different methods for developing a decellularized arterial scaffold. The first uses an electrode clamp that is applied to the outside of a rodent carotid artery and the second applies an endovascular minimally invasive approach to apply electric fields from the inner surface of the blood vessels. Both methods are first modeled using a transient finite element analysis of electric and thermal fields to ensure that the electric parameters used in this study will result in minimal thermal damage. Experimental work demonstrates that both techniques result in not only a decellularized arterial construct but an endothelial regrowth is evident along the lumen 7 days after treatment, indicating that the extracellular matrix was not damaged by electric and thermal fields and is still able to support cell growth. [ABSTRACT FROM AUTHOR]

Intense but short electrical fields can increase the permeability of the cell membrane in a process referred to as electroporation. Reversible electroporation has become an important tool in biotechnology and medicine. The various applications of reversible electroporation require cells to survive the procedure, and therefore the occurrence of irreversible electroporation (IRE), following which cells die, is obviously undesirable. However, for the past few years, IRE has begun to emerge as an important minimally invasive nonthermal ablation technique in its own right as a method to treat tumors and arrhythmogenic regions in the heart. IRE had been studied primarily to define the upper limit of electrical parameters that induce reversible electroporation. Thus, the delineation of IRE from thermal damage due to Joule heating has not been thoroughly investigated. The goal of this study was to express the upper bound of IRE (onset of thermal damage) theoretically as a function of physical properties and electrical pulse parameters. Electrical pulses were applied to THP-1 human monocyte cells, and the percentage of irreversibly electroporated (dead) cells in the sample was quantified. We also determined the upper bound of IRE (onset of thermal damage) through a theoretical calculation that takes into account the physical properties of the sample and the electric pulse characteristics. Our experimental results were achieved below the theoretical curve for the onset of thermal damage. These results confirm that the region to induce IRE without thermal damage is substantial. We believe that our new theoretical analysis will allow researchers to optimize IRE parameters without inducing deleterious thermal effects. [ABSTRACT FROM AUTHOR]

Nonthermal irreversible electroporation (NTIRE) is a new minimally invasive surgical technique that is part of the emerging field of molecular surgery, which holds the potential to treat diseases with unprecedented accuracy. NTIRE utilizes electrical pulses delivered to a targeted area, producing irreversible damage to the cell membrane. Because NTIRE does not cause thermal damage, the integrity of all other molecules, collagen, and elastin in the targeted area is preserved. Previous theoretical studies have only examined NTIRE in homogeneous tissues; however, biological structures are complex collections of diverse tissues. In order to develop electroporation as a precise treatment in clinical applications, realistic models are necessary. Therefore, the purpose of this study was to refine electroporation as a treatment by examining the effect of NTIRE in heterogeneous tissues of the prostate and breast. This study uses a two-dimensional finite element solution of the Laplace and bioheat equations to examine the effects of heterogeneities on electric field and temperature distribution. Three different heterogeneous structures were taken into account: nerves, blood vessels, and ducts. The results of this study demonstrate that heterogeneities significantly impact both the temperature and electrical field distribution in surrounding tissues, indicating that heterogeneities should not be neglected. The results were promising. While the surrounding tissue experienced a high electrical field, the axon of the nerve, the interior of the blood vessel, and the ducts experienced no electrical field. This indicates that blood vessels, nerves, and lactiferous ducts adjacent to a tumor treated with electroporation will survive, while the cancerous lesion is ablated. This study clearly demonstrates the importance of considering heterogeneity in NTIRE applications. [ABSTRACT FROM AUTHOR]