A micro circulating PCR chip using a suction-type membrane for fluidic transport

A new micromachined circulating polymerase chain reaction (PCR) chip is reported in this study. A novel liquid transportation mechanism utilizing a suction-type membrane and three microvalves were used to create a new microfluidic control module to rapidly transport the DNA samples and PCR reagents around three bio-reactors operating at three different temperatures. When operating at a membrane actuation frequency of 14.29 Hz and a pressure of 5 psi, the sample flow rate in the microfluidic control module can be as high as 18 μL/s. In addition, an array-type microheater was adopted to improve the temperature uniformity in the reaction chambers. Open-type reaction chambers were designed to facilitate temperature calibration. Experimental data from infrared images showed that the percentage of area inside the reaction chamber with a thermal variation of less than 1°C was over 90% for a denaturing temperature of 94°C. Three array-type heaters and temperature sensors were integrated into this new circulating PCR chip to modulate three specific operating temperatures for the denaturing, annealing, and extension steps of a PCR process. With this approach, the cycle numbers and reaction times of the three separate reaction steps can be individually adjusted. To verify the performance of this circulating PCR chip, a PCR process to amplify a detection gene (150 base pairs) associated with the hepatitis C virus was performed. Experimental results showed that DNA samples with concentrations ranging from 10⁵ to 10²copies/μL can be successfully amplified. Therefore, this new circulating PCR chip may provide a useful platform for genetic identification and molecular diagnosis.     

双激励线圈磁感应断层成像的可行性研究

研究磁感应断层成像中两个激励线圈相对于单个激励线圈时增强被测对象内部磁感应强度的可行性。利用电磁场数值计算软件Comsol Multiphysics,在三维球形头模型外周设置单个激励线圈或呈不同夹角的两个激励线圈(夹角分别设置为30°、60°、90°、120°、150°、180°),分别计算中心有病变组织和无病变组织时中心位置的磁感应强度,并加以比较。中心无病变组织时,除夹角为180°外,中心位置的磁感应强度约为单激励线圈的2倍;中心有病变组织时,除夹角为180°外,随夹角增大中心位置的磁感应强度从单激励线圈的2倍开始逐渐增大;除夹角为180°外,有病变和无病变时中心位置的磁感应强度的变化量随夹角的增大而逐渐减小,但是夹角小于90°时该变化量大于单激励线圈的变化量。两激励线圈夹角为180°时,无论中心有无病变组织,中心位置的磁感应强度均减小。结果表明:双激励线圈较单激励线圈有可能增大被测对象内的磁感应强度;通过合理选择双激励线圈的夹角,有可能放大被测对象内部病变组织导致的磁感应强度的变化;两者均可改善磁感应断层成像。     

Effect of actuation sequence on flow rates of peristaltic micropumps with PZT actuators

Many biomedical applications require the administration of drugs at a precise and preferably programmable rate. The flow rate generated by the peristaltic micropumps used in such applications depends on the actuation sequence. Accordingly, the current study performs an analytical and experimental investigation to determine the correlation between the dynamic response of the diaphragms in the micropump and the actuation sequence. A simple analytical model of a peristaltic micropump is established to analyze the shift in the resonant frequency of the diaphragms caused by the viscous damping effect. The analytical results show that this damping effect increases as the oscillation frequency of the diaphragm increases. A peristaltic micropump with three piezoelectric actuators is fabricated on a silicon substrate and is actuated using 2-, 3-, 4- and 6-phase actuation sequences via a driving system comprising a microprocessor and a phase controller. A series of experiments is conducted using de-ionized water as the working fluid to determine the diaphragm displacement and the flow rates induced by each of the different actuation sequences under phase frequencies ranging from 50 Hz to 1 MHz. The results show that the damping effect of actuation sequences influences diaphragm resonant frequency, which in turn affects the profiles of flow rates.     

Flow-orthogonal bead oscillation in a microfluidic chip with a magnetic anisotropic flux-guide array

A new concept for the manipulation of superparamagnetic beads inside a microfluidic chip is presented in this paper. The concept allows for bead actuation orthogonal to the flow direction inside a microchannel. Basic manipulation functionalities were studied by means of finite element simulations and results were oval-shaped steady state oscillations with bead velocities up to 500 μm/s. The width of the trajectory could be controlled by prescribing external field rotation. Successful verification experiments were performed on a prototype chip fabricated with excimer laser ablation in polycarbonate and electroforming of nickel flux-guides. Bead velocities up to 450 μm/s were measured in a 75 μm wide channel. By prescribing the currents in the external quadrupole magnet, the shape of the bead trajectory could be controlled.     

Microneedle Insertion Force Reduction Using Vibratory Actuation

The effect of vibratory actuation on microneedle insertion force was investigated. Hollow micro hypodermic injection needles were fabricated by a two-wafer polysilicon micromolding process. A vibratory actuator operating in the kHz range was coupled with the hypodermic microneedles. The force to insert microneedles into excized animal tissue was measured with a load cell. Results showed a greater than 70% reduction in microneedle insertion force by using vibratory actuation. The application of vibratory actuation provides a promising method to precisely control the microneedle insertion forces to overcome microneedle structural material limitations, minimize insertion pain, and enhance the efficiency of drug delivery.     

Magnetophosphenes: a quantitative analysis of thresholds

Low-frequency and transient magnetic fields of moderate flux densities are known to generate visual phenomena, so-called magnetophosphenes. In the present study, time-variable very low frequency (10–50 Hz) electromagnetic fields of moderate flux density (0–40 mT) were used to induce magnetophosphenes. The threshold values for these phosphenes were determined as a function of the frequency of the magnetic field both in normal subjects and colour defective ones. Maximum sensitivity occurred at a frequency of approximately 20–30 Hz, and with broad-spectrum light the threshold flux density was 10–12 mT. The threshola values were found to be dependent upon the intensity and the spectral distribution of the background light. Sensitivity decreased during dark adaptation. In certain respects deutans differed from subjects with normal colour vision. Possible mechanisms for generation of magnetophosphenes are discussed. The present magnetic threshold curves show a close resemblance to corresponding curves obtained by electric stimulation at various frequencies provided the electric thresholds are divided by the a.c. frequency. These problems are under current investigation in our laboratory. This is in full agreement with the assumption that the fluctuating magnetic field affects retinal neurons by inducing currents which polarise synaptic terminals.     

Sensing magnetic flux density of artificial neurons with a MEMS device

We describe a simple procedure to characterize a magnetic field sensor based on microelectromechanical systems (MEMS) technology, which exploits the Lorentz force principle. This sensor is designed to detect, in future applications, the spiking activity of neurons or muscle cells. This procedure is based on the well-known capability that a magnetic MEMS device can be used to sense a small magnetic flux density. In this work, an electronic neuron (FitzHugh–Nagumo) is used to generate controlled spike-like magnetic fields. We show that the magnetic flux density generated by the hardware of this neuron can be detected with a new MEMS magnetic field sensor. This microdevice has a compact resonant structure (700 × 600 × 5 μm) integrated by an array of silicon beams and p-type piezoresistive sensing elements, which need an easy fabrication process. The proposed microsensor has a resolution of 80 nT, a sensitivity of 1.2 V⋅T⁻¹, a resonant frequency of 13.87 kHz, low power consumption (2.05 mW), quality factor of 93 at atmospheric pressure, and requires a simple signal processing circuit. The importance of our study is twofold. First, because the artificial neuron can generate well-controlled magnetic flux density, we suggest it could be used to analyze the resolution and performance of different magnetic field sensors intended for neurobiological applications. Second, the introduced MEMS magnetic field sensor may be used as a prototype to develop new high-resolution biomedical microdevices to sense magnetic fields from cardiac tissue, nerves, spinal cord, or the brain.     

Kinematics and flow characteristics of a magnetic actuated multi-cilia configuration

The current paper continues the analysis of a completely novel method of fluid manipulation technology in micro-fluidics systems, inspired by nature, namely by the mechanisms found in ciliates. More information on this subject can be found at http://www.hitech-projects.com/euprojects/artic/. In order to simulate the drag forces acting on an array of artificial cilia, we have developed a computer code that is based on fundamental solutions of Stokes flow in a semi-infinite domain. The actuation mechanism consists of a bi-directional rotating excitation magnetic field. The magnetization induced by the magnetic field was calculated in a separate routine based on the Integral Nonlinear Equations Approach with 1D discretization of wire (cilium). Time averaged x-coordinate mass flow rates, streamlines and vorticity field are computed for several cilium configurations. The outcome and originality of this paper consist on assessing magnetic actuation as a practical tool for obtaining a consistent one-directional fluid flow.     

Reconstruction of conductivity using the dual-loop method with one injection current in MREIT

Magnetic resonance electrical impedance tomography (MREIT) is to visualize the internal current density and conductivity of an electrically conductive object. Injecting current through surface electrodes, we measure one component of the induced internal magnetic flux density using an MRI scanner. In order to reconstruct the conductivity distribution inside the imaging object, most algorithms in MREIT have required multiple magnetic flux density data by injecting at least two independent currents. In this paper, we propose a direct method to reconstruct the internal isotropic conductivity with one component of magnetic flux density data by injecting one current into the imaging object through a single pair of surface electrodes. Firstly, the proposed method reconstructs a projected current density which is a uniquely determined current from the measured one-component magnetic flux density. Using a relation between voltage potential and current, based on Kirchhoff's voltage law, the proposed method is designed to use a combination of two loops around each pixel from which to derive an implicit matrix system for determination of the internal conductivity. Results from numerical simulations demonstrate that the proposed algorithm stably determines the conductivity distribution in an imaging slice. We compare the reconstructed internal conductivity distribution using the proposed method with that using a conventional method with agarose gel phantom experiments.     

Locating steel needles in the human body using a SQUID magnetometer

A technique has been developed, based on magnetic field measurements, to localize, in three dimensions, hypodermic and sewing needles lost in the human body. A theoretical model for the magnetic field generated by needles has been elaborated and experimentally validated. Using this model, the localization technique gives information about needle's centre, orientation and depth. The clinical measurements have been made using a SQUID system, with patients being moved under the sensor with the aid of an X-Y bed. The magnetic field associated with the remanent magnetization of the needle is acquired on-line and mapped over a plane. In all six cases that occurred, the technique allowed surgical localization of the needles with ease and high precision. This procedure can decrease the surgery time for extraction of foreign bodies by a large factor, and also reduce the generally high odds of failure.     

Improved conductivity reconstruction from multi-echo MREIT utilizing weighted voxel-specific signal-to-noise ratios

Magnetic resonance electrical impedance tomography (MREIT) is a non-invasive method to visualize cross-sectional electrical conductivity and/or current density by measuring a magnetic flux density signal when an electrical current is injected into a subject. In the MREIT system, it is crucial to reduce the scan duration while maintaining spatial resolution and contrast for practical in vivo implementation. The purpose of the study is to optimize the measured magnetic flux density using an interleaved multiple-echo pulse sequence (injected current nonlinear encoding) that acquires each spatial position multiple times, although these pixels vary between echoes in their signal-to-noise ratio due to (a) T*2 decay and (b) the current density passing through the pixel. Using the acquired multiple measured magnetic flux densities, the noise level for the measured magnetic flux density B(z) at each pixel is estimated using the relationship between the intensity of the magnitude and the width of the injected current. We determine an optimal combination of the multiply acquired magnetic flux densities, which optimally reduces the random noise artifacts. We develop a new denoising technique and apply it to a recovered conductivity distribution with a known noise level of the recovered magnetic flux density, which is designed to provide a stable internal conductivity distribution, while sustaining resolution. The proposed method uses three key steps: the first step is optimizing the magnetic flux density by using the multiple-echo magnetic flux densities at each pixel, the second step is estimating the noise level of this optimized magnetic flux density and the third step is applying a denoising technique using the pixel-specific estimated noise level. Numerical simulation experiments using a three-dimensional cylindrical phantom model validated the proposed method. Multiple-echo B(z) data were generated, including in short T*2 or low spin-density regions, as a function of T*2 and the temporal extent of the injected current. In the simulation experiment, comparing between an equally averaged and the optimized B(z) methods, relative L2-mode errors were 0.053 and 0.024, respectively. In an actual imaging experiment of an agarose gel filled with objects of various conductivities and shapes, we acquired six echoes per repetition time. The optimal weighting factors minimized the effects of noise in B(z), and provided reconstructed conductivity maps that suppressed noise artifacts that normally accumulate in the low signal-to-noise-ratio defect regions.     

A field effect transistor (FET)-based immunosensor for detection of HbA1c and Hb

A field effect transistor (FET)-based immunosensor was developed for diabetes monitoring by detecting the concentrations of glycated hemoglobin (HbA1c) and hemoglobin (Hb). This immunosensor consists of a FET-based sensor chip and a disposable extended-gate electrode chip. The sensor chip was fabricated by standard CMOS process and was integrated with signal readout circuit. The disposable electrode chip, fabricated on polyester plastic board by Micro-Electro-Mechanical-Systems (MEMS) technique, was integrated with electrodes array and micro reaction pool. Biomolecules were immobilized on the electrode based on self-assembled monolayer and gold nanoparticles. Experimental results showed that the immunosensor achieved a linear response to HbA1c with the concentration from 4 to 24 μg/ml, and a linear response to Hb with the concentration from 60 to 180 μg/ml.     

A biopsy tool with integrated piezoceramic elements for needle tract cauterization and cauterization monitoring

This paper reports the feasibility of biopsy needle tract cauterization and cauterization monitoring using an embedded array of piezoceramic microheaters. Circular heaters of lead zirconate titanate (PZT-5A), with 200 μm diameter and 70–80 μm thickness, are fabricated using a batch mode micro ultrasonic machining process. These are then assembled into cavities in the walls of 20-gauge stainless steel needles and sealed with epoxy. Experiments are performed by inserting the proposed biopsy needle into porcine tissue samples. The needle surface exceeds the minimum target temperature rise of 33°C for either radial or thickness mode vibrations. The corresponding input power levels are 236 mW and 325 mW, respectively. The tissue cauterization extends 1–1.25 mm beyond the perimeter of the needle and is uniform in all directions. After cauterization, the fundamental anti-resonance frequency and the corresponding impedance magnitude of the PZT heater decrease by 4.1% and 42.6%, respectively, thereby providing a method to monitor the extent of tissue cauterization. A sensing interface circuit capable of measuring the resonance frequency shift of the PZT elements is built and tested using discrete integrated circuit components. The circuit detects the resonance frequency shift from 8.22 MHz to 7.96 MHz of the PZT elements when the biopsy needle is inserted into wax medium. An interface circuit for actuation of the PZT elements for tissue cauterization is also described.     

Enhanced endothelial activity reflected in cutaneous blood flow oscillations of athletes

Functional alterations of vascular endothelial cells may be evaluated by analysing differences in effects of endothelium-dependent [acetylcholine (ACh)] and endothelium-independent [sodium nitroprusside (SNP)] vasodilators. We evaluated whether a dynamic approach using spectral analysis of the blood flow signal, resulting from the cutaneous red cell flux and recorded by the technique of laser Doppler flowmetry (LDF), can detect higher endothelial responsiveness in trained versus less trained individuals. There was a 1.6 times higher ACh-induced cutaneous perfusion in athletes than in controls (P<0.05), both when evaluated as a mean value of the LDF signal or as the amplitudes of its spectral components. In the frequency interval from 0.009 to 1.6 Hz, ACh induced a 1.6 times higher average spectral amplitude (P<0.01) in athletes compared with controls. ACh also induced a 1.6 times higher absolute spectral amplitude of the oscillator at around 0.01 Hz (P<0.05) in the athletes compared with the controls, whereas the endothelial oscillation at around 0.01 Hz during basal unstimulated perfusion was 1.5 times higher (P<0.01). There were no significant differences in absolute or relative amplitude during iontophoresis with SNP. These results indicate that athletes have higher endothelial activity than less trained individuals.     

Integrated polymerase chain reaction chips utilizing digital microfluidics

This study reports an integrated microfluidic chip for polymerase chain reaction (PCR) applications utilizing digital microfluidic chip (DMC) technology. Several crucial procedures including sample transportation, mixing, and DNA amplification were performed on the integrated chip using electro-wetting-on-dielectric (EWOD) effect. An innovative concept of hydrophobic/hydrophilic structure has been successfully demonstrated to integrate the DMC chip with the on-chip PCR device. Sample droplets were generated, transported and mixed by the EWOD-actuation. Then the mixture droplets were transported to a PCR chamber by utilizing the hydrophilic/hydrophobic interface to generate required surface tension gradient. A micro temperature sensor and two micro heaters inside the PCR chamber along with a controller were used to form a micro temperature control module, which could perform precise PCR thermal cycling for DNA amplification. In order to demonstrate the performance of the integrated DMC/PCR chips, a detection gene for Dengue II virus was successfully amplified and detected. The new integrated DMC/PCR chips only required an operation voltage of 12V_RMS at a frequency of 3 KHz for digital microfluidic actuation and 9V_DC for thermal cycling. When compared to its large-scale counterparts for DNA amplification, the developed system consumed less sample and reagent and could reduce the detection time. The developed chips successfully demonstrated the feasibility of Lab-On-a-Chip (LOC) by utilizing EWOD-based digital microfluidics.     

Hysteretic Buckling for Actuation of Magnet–Polymer Composites

The development of bioinspired, novel, high‐performance, “artificial muscle” materials is a topic of high current interest. Flexibility for folding and actuation are key parameters in the development of such artificial muscles, especially for self‐assembly and origami engineering. A current challenge is to develop a multifunctional material that combines the biological functions of actuation, sensing, and healing. In this work, for the first time experimental and modeling results of the actuation capability of a multifunctional magnet–polymer (Magpol) composite material that is already capable of damage sensing and self‐healing are reported. Actuation by constrained buckling of Magpol in an external magnetic field shows two buckling modes. The stress and strain are studied and optimized for various actuator geometries. Work density of up to 16 J kg^?1 is obtained, comparable to electroactive polymers. Magpol is successfully able to lift more than four times its own weight at relatively low applied magnetic field. Good agreement is observed between the experimental and modeling results. An Ashby design chart is employed to compare its performance to other actuators. This work provides insight into the development of next generation flexible, active multifunctional materials.

     

Effects of 50 Hz electromagnetic fields on electroencephalographic alpha activity, dental pain threshold and cardiovascular parameters in humans

Recent studies indicate that exposure to extremely low frequency magnetic fields (ELF MFs) influences human electroencephalographic (EEG) alpha activity and pain perception. In the present study we analyse the effect on electrical EEG activity in the alpha band (8-13 Hz) and on nociception in 40 healthy male volunteers after 90-min exposure of the head to 50 Hz ELF MFs at a flux density of 40 or 80 microT in a double-blind randomized sham-controlled study. Since cardiovascular regulation is functionally related to pain modulation, we also measured blood pressure (BP) and heart rate (HR) during treatment. Alpha activity after 80 microT magnetic treatment almost doubled compared to sham treatment. Pain threshold after 40 microT magnetic treatment was significantly lower than after sham treatment. No effects were found for BP and HR. We suggest that these results may be explained by a modulation of sensory gating processes through the opioidergic system, that in turn is influenced by magnetic exposure.     

Implantable drug delivery device using frequency-controlled wireless hydrogel microvalves

This paper reports a micromachined drug delivery device that is wirelessly operated using radiofrequency magnetic fields for implant applications. The controlled release from the drug reservoir of the device is achieved with the microvalves of poly(N-isopropylacrylamide) thermoresponsive hydrogel that are actuated with a wireless resonant heater, which is activated only when the field frequency is tuned to the resonant frequency of the heater circuit. The device is constructed by bonding a 1-mm-thick polyimide component with the reservoir cavity to the heater circuit that uses a planar coil with the size of 5–10 mm fabricated on polyimide film, making all the outer surfaces to be polyimide. The release holes created in a reservoir wall are opened/closed by the hydrogel microvalves that are formed inside the reservoir by in-situ photolithography that uses the reservoir wall as a photomask, providing the hydrogel structures self-aligned to the release holes. The wireless heaters exhibit fast and strong response to the field frequency, with a temperature increase of up to 20°C for the heater that has the 34-MHz resonant frequency, achieving 38-% shrinkage of swelled hydrogel when the heater is excited at its resonance. An active frequency range of ~2 MHz is observed for the hydrogel actuation. Detailed characteristics in the fabrication and actuation of the hydrogel microvalves as well as experimental demonstrations of frequency-controlled temporal release are reported.     

Hydrogel swelling as a trigger to release biodegradable polymer microneedles in skin

Biodegradable polymeric microneedles were developed as a method for achieving sustained transdermal drug release. These microneedles have potential as a patient-friendly substitute for conventional sustained release methods. However, they have limitations related to the difficulty of achieving separation of the needles into the skin. We demonstrated that microneedle separation into the skin was mediated by hydrogel swelling in response to contact with body fluid after the needles were inserted into the skin. The hydrogel microparticles were synthesized by an emulsification method using poly-N-isopropylacrylamide (PNIPAAm). The microneedles were fabricated by micromolding poly-lactic-co-glycolic acid (PLGA) after filling the cavities of the mold with the hydrogel microparticles. The failure of microneedle tips caused by hydrogel swelling was studied in regard to contact with water, insertion of microneedles into porcine cadaver skin in vitro, stress-strain behavior, and insertion into the back skin of a hairless mouse in vivo. The drug delivery property of the hydrogel particles was investigated qualitatively by inserting polymer microneedles into porcine cadaver skin in vitro, and the sustained release property of PLGA microneedles containing hydrogel microparticles was studied quantitatively using the Franz cell model. The hydrogel particles absorbed water quickly, resulting in the cracking of the microneedles due to the difference in volume expansion between the needle matrix polymer and the hydrogel particles. The swollen particles caused the microneedles to totally breakdown, leaving the microneedle tips in the porcine cadaver skin in vitro and in the hairless mouse skin in vivo. Model drugs encapsulated in biodegradable polymer microneedles and hydrogel microparticles were successfully delivered by releasing microneedles into the skin.     

脉部磁场对细胞缝隙连接通讯功能影响的研究

本文用细胞内微注射法对不同强度的脉冲磁场对细胞缝隙连接通讯（GJIC）功能的效应进行了研究,并与正弦磁场的作用以及脉冲磁场所感生的不同电场的作用进行了比较和探索。研究发现0.41mT以上强度的脉冲磁场具有抑制GJIC的作用。这种作用与磁场强度有关,其抑制GJIC的效庆比正弦磁场为强。经相同磁场不同强度感应电场作用的比较。得出,GJIC的抑制主要由磁场引起,而与感应电场关系不大,本文首次从研究对GJIC的作用提供了脉冲磁场治疗的创伤愈合等有关理论依据。