Frequency-Resolved High-Frequency Broadband Measurement of Acoustic Longitudinal Waves by Laser-Based Excitation and Detection.

Optoacoustics is a metrology widely used for material characterisation. In this study, a measurement setup for the selective determination of the frequency-resolved phase velocities and attenuations of longitudinal waves over a wide frequency range (3-55 MHz) is presented. The ultrasonic waves in this setup were excited by a pulsed laser within an absorption layer in the thermoelastic regime and directed through a layer of water onto a sample. The acoustic waves were detected using a self-built adaptive interferometer with a photorefractive crystal. The instrument transmits compression waves only, is low-contact, non-destructive, and has a sample-independent excitation. The limitations of the approach were studied both by simulation and experiments to determine how the frequency range and precision can be improved. It was shown that measurements are possible for all investigated materials (silicon, silicone, aluminium, and water) and that the relative error for the phase velocity is less than 0.2%.

Guided Acoustic Waves in Polymer Rods with Varying Immersion Depth in Liquid.

Monitoring tanks and vessels play an important part in public infrastructure and several industrial processes. The goal of this work is to propose a new kind of guided acoustic wave sensor for measuring immersion depth. Common sensor types such as pressure sensors and airborne ultrasonic sensors are often limited to non-corrosive media, and can fail to distinguish between the media they reflect on or are submerged in. Motivated by this limitation, we developed a guided acoustic wave sensor made from polyethylene using piezoceramics. In contrast to existing sensors, low-frequency Hanning-windowed sine bursts were used to excite the L(0,1) mode within a solid polyethylene rod. The acoustic velocity within these rods changes with the immersion depth in the surrounding fluid. Thus, it is possible to detect changes in the surrounding media by measuring the time shifts of zero crossings through the rod after being reflected on the opposite end. The change in time of zero crossings is monotonically related to the immersion depth. This relative measurement method can be used in different kinds of liquids, including strong acids or bases.

Quantification of Humidity and Salt Detection in Historical Building Materials via Broadband Radar Measurement.

For the investigation of moisture and salt content in historic masonry, destructive drilling samples followed by a gravimetric investigation is still the preferred method. In order to prevent the destructive intrusion into the building substance and to enable a large-area measurement, a nondestructive and easy-to-use measuring principle is needed. Previous systems for moisture measurement usually fail due to a strong dependence on contained salts. In this work, a ground penetrating radar (GPR) system was used to determine the frequency-dependent complex permittivity in the range between 1 and 3 GHz on salt-loaded samples of historical building materials. By choosing this frequency range, it was possible to determine the moisture in the samples independently of the salt content. In addition, it was possible to make a quantitative statement about the salt level. The applied method demonstrates that with ground penetrating radar measurements in the frequency range selected here, a salt-independent moisture determination can be carried out.

Ultrasonic Interferometric Procedure for Quantifying the Bone-Implant Interface.

The loosening of an artificial joint is a frequent and critical complication in orthopedics and trauma surgery. Due to a lack of accuracy, conventional diagnostic methods such as projection radiography cannot reliably diagnose loosening in its early stages or detect whether it is associated with the formation of a biofilm at the bone-implant interface. In this work, we present a non-invasive ultrasound-based interferometric measurement procedure for quantifying the thickness of the layer between bone and prosthesis as a correlate to loosening. In principle, it also allows for the material characterization of the interface. A well-known analytical model for the superposition of sound waves reflected in a three-layer system was combined with a new method in data processing to be suitable for medical application at the bone-implant interface. By non-linear fitting of the theoretical prediction of the model to the actual shape of the reflected sound waves in the frequency domain, the thickness of the interlayer can be determined and predictions about its physical properties are possible. With respect to determining the layer's thickness, the presented approach was successfully applied to idealized test systems and a bone-implant system in the range of approx. 200 µm to 2 mm. After further optimization and adaptation, as well as further experimental tests, the procedure offers great potential to significantly improve the diagnosis of prosthesis loosening at an early stage and may also be applicable to detecting the formation of a biofilm.

On Dispersion Compensation for GAW-Based Structural Health Monitoring.

Guided acoustic waves (GAW) have proven to be a useful tool for structural health monitoring (SHM). However, the dispersive nature of commonly used Lamb waves compromises the spatial resolution making it difficult to detect small or weakly reflective defects. Here we demonstrate an approach that can compensate for the dispersive effects, allowing advanced algorithms to be used with significantly higher signal-to-noise ratio and spatial resolution. In this paper, the sign coherence factor (SCF) extension of the total focusing method (TFM) algorithm is used. The effectiveness is examined by numerical simulation and experimentally demonstrated by detecting weakly reflective layers with a highly dispersive A0 mode on an aluminum plate, which are not detectable without compensating for the dispersion effects.

Acoustic Limescale Layer and Temperature Measurement in Ultrasonic Flow Meters.

Guided acoustic waves are commonly used in domestic water meters to measure the flow rate. The accuracy of this measurement method is affected by factors such as variations in temperature and limescale deposition inside of the pipe. In this work, a new approach using signals from different sound propagation paths is used to determine these quantities and allow for subsequent compensation. This method evaluates the different propagation times of guided Lamb waves in flow measurement applications. A finite element method-based model is used to identify the calibration curves for the device under test. The simulated dependencies on temperature and layer thickness are validated by experimental data. Finally, a test on simulated data with varying temperatures and limescale depositions proves that this method can be used to separate both effects. Based on these values, a flow measurement correction scheme can be derived that provides an improved resolution of guided acoustic wave-based flow meters.

Elastic Properties Measurement Using Guided Acoustic Waves.

Nondestructive evaluation of elastic properties plays a critical role in condition monitoring of thin structures such as sheets, plates or tubes. Recent research has shown that elastic properties of such structures can be determined with remarkable accuracy by utilizing the dispersive nature of guided acoustic waves propagating in them. However, existing techniques largely require complicated and expensive equipment or involve accurate measurement of an additional quantity, rendering them impractical for industrial use. In this work, we present a new approach that requires only a pair of piezoelectric transducers used to measure the group velocities ratio of fundamental guided wave modes. A numerical model based on the spectral collocation method is used to fit the measured data by solving a bound-constrained nonlinear least squares optimization problem. We verify our approach on both simulated and experimental data and achieve accuracies similar to those reported by other authors. The high accuracy and simple measurement setup of our approach makes it eminently suitable for use in industrial environments.

[Lab on a Chip]. / "Lab on a Chip".

BACKGROUND: Miniaturization has not only driven microelectronics and generated new unforeseen options but has also dramatically changed sensors and analytics. DEVELOPMENTS: The Lab on a Chip (LOC) technology enables laboratory processes to run fully automated in canals in the micrometre range. The biggest challenge for LOC is to keep production costs low despite miniaturization and application-specific design. If this is achieved medical laboratory analyses can usually be carried out faster and with less hands on time. This explains why LOCs are already integrated into many laboratory instruments and why point-of-care testing (POCT) can no longer be imagined without it. New markers, such as in liquid biopsies and measurement techniques, such as Raman spectroscopy and mass spectroscopy, create further potentials that will enable faster and more specific laboratory analyses to be made using LOC technology. CONCLUSION: The LOC technology has the potential of changing the medical practice especially in cases when the central laboratory is not available or is unable to provide results fast enough.

Monitoring of Soft Deposition Layers in Liquid-Filled Tubes with Guided Acoustic Waves Excited by Clamp-on Transducers.

The monitoring of liquid-filled tubes with respect to the formation of soft deposition layers such as biofilms on the inner walls calls for non-invasive and long-term stable sensors, which can be attached to existing pipe structures. For this task a method is developed, which uses an ultrasonic clamp-on device. This method is based on the impact of such deposition layers on the propagation of circumferential guided waves on the pipe wall. Such waves are partly converted into longitudinal compressional waves in the liquid, which are back-converted to guided waves in a circular cross section of the pipe. Validating this approach, laboratory experiments with gelatin deposition layers on steel tubes exhibited a distinguishable sensitivity of both wave branches with respect to the thickness of such layers. This allows the monitoring of the layer growth.

Modeling size controlled nanoparticle precipitation with the co-solvency method by spinodal decomposition.

The co-solvency method is a method for the size controlled preparation of nanoparticles like polymersomes, where a poor co-solvent is mixed into a homogeneous copolymer solution to trigger precipitation of the polymer. The size of the resulting particles is determined by the rate of co-solvent addition. We use the Cahn-Hilliard equation with a Flory-Huggins free energy model to describe the precipitation of a polymer under changing solvent quality by applying a time dependent Flory-Huggins interaction parameter. The analysis focuses on the characteristic size R of polymer aggregates that form during the initial spinodal decomposition stage, and especially on how R depends on the rate s of solvent quality change. Both numerical results and a perturbation analysis predict a power law dependence R∼s(-⅙), which is in agreement with power laws for the final particle sizes that have been reported from experiments and molecular dynamics simulations. Hence, our model results suggest that the nanoparticle size in size-controlled precipitation is essentially determined during the spinodal decomposition stage.

Integrated DNA and RNA extraction and purification on an automated microfluidic cassette from bacterial and viral pathogens causing community-acquired lower respiratory tract infections.

In this paper, we describe the development of an automated sample preparation procedure for etiological agents of community-acquired lower respiratory tract infections (CA-LRTI). The consecutive assay steps, including sample re-suspension, pre-treatment, lysis, nucleic acid purification, and concentration, were integrated into a microfluidic lab-on-a-chip (LOC) cassette that is operated hands-free by a demonstrator setup, providing fluidic and valve actuation. The performance of the assay was evaluated on viral and Gram-positive and Gram-negative bacterial broth cultures previously sampled using a nasopharyngeal swab. Sample preparation on the microfluidic cassette resulted in higher or similar concentrations of pure bacterial DNA or viral RNA compared to manual benchtop experiments. The miniaturization and integration of the complete sample preparation procedure, to extract purified nucleic acids from real samples of CA-LRTI pathogens to, and above, lab quality and efficiency, represent important steps towards its application in a point-of-care test (POCT) for rapid diagnosis of CA-LRTI.

Fast nucleic acid amplification for integration in point-of-care applications.

An ultrafast microfluidic PCR module (30 PCR cycles in 6 min) based on the oscillating fluid plug concept was developed. A robust amplification of native genomic DNA from whole blood samples could be achieved at operational conditions established from systematic investigations of key parameters including heat transfer and in particular flow velocities. Experimental data were augmented with results from computational fluid dynamics simulations. The reproducibility of the current system was substantially improved compared to previous concepts by integration of a closed reservoir instead of utilizing a vented channel end at ambient pressure rendering the devised module suitable for integration into complex sample-to-answer analysis platforms such as point-of-care applications.

Towards a "Sample-In, Answer-Out" Point-of-Care Platform for Nucleic Acid Extraction and Amplification: Using an HPV E6/E7 mRNA Model System.

The paper presents the development of a "proof-of-principle" hands-free and self-contained diagnostic platform for detection of human papillomavirus (HPV) E6/E7 mRNA in clinical specimens. The automated platform performs chip-based sample preconcentration, nucleic acid extraction, amplification, and real-time fluorescent detection with minimal user interfacing. It consists of two modular prototypes, one for sample preparation and one for amplification and detection; however, a common interface is available to facilitate later integration into one single module. Nucleic acid extracts (n = 28) from cervical cytology specimens extracted on the sample preparation chip were tested using the PreTect HPV-Proofer and achieved an overall detection rate for HPV across all dilutions of 50%-85.7%. A subset of 6 clinical samples extracted on the sample preparation chip module was chosen for complete validation on the NASBA chip module. For 4 of the samples, a 100% amplification for HPV 16 or 33 was obtained at the 1 : 10 dilution for microfluidic channels that filled correctly. The modules of a "sample-in, answer-out" diagnostic platform have been demonstrated from clinical sample input through sample preparation, amplification and final detection.

Justification of rapid prototyping in the development cycle of thermoplastic-based lab-on-a-chip.

During the developmental cycle of lab-on-a-chip devices, various microstructuring techniques are required. While in the designing and assay implementation phase direct structuring or so-called rapid-prototyping methods such as milling or laser ablation are applied, replication methods like hot embossing or injection moulding are favourable for large quantity manufacturing. This work investigated the applicability of rapid-prototyping techniques for thermoplastic chip development in general, and the reproducibility of performances in dependency of the structuring technique. A previously published chip for prenatal diagnosis that preconcentrates DNA via electrokinetic trapping and field-amplified-sample-stacking and afterwards separates it in CGE was chosen as a model. The impact of structuring, sealing, and the integration of membranes on the mobility of the EOF, DNA preconcentration, and DNA separation was studied. Structuring methods were found to significantly change the location where preconcentration of DNA occurs. However, effects on the mobility of the EOF and the separation quality of DNA were not observed. Exchange of the membrane has no effect on the chip performance, whereas the sealing method impairs the separation of DNA within the chip. The overall assay performance is not significantly influenced by different structuring methods; thus, the application of rapid-prototyping methods during a chip development cycle is well justified.

On-chip analysis of respiratory viruses from nasopharyngeal samples.

Point-of-care (PoC) testing followed by personalized efficient therapy of infectious diseases may result in a considerable reduction of associated health care costs. Lab-on-a-chip (LoC) systems represent a potentially high efficient class of PoC tools. Here, we present a LoC system for automated pathogen analysis of respiratory viruses from nasopharyngeal specimens. The device prepares total nucleic acids from extracted swab samples using magnetic silica beads. After reverse transcription the co-purified viral RNA is amplified in accordance with the QIAplex multiplex PCR technology. Hybridized to corresponding QIAGEN LiquiChip beads and labelled with streptavidin R-phycoerythrin, the amplified target sequences are finally detected using a QIAGEN LiquiChip200 workstation. All chemicals needed are either stored freeze-dried on the disposable chip or are provided in liquid form in a reagent cartridge for up to 24 runs. Magnetic stir bars for mixing as well as turning valves with metering structures are integrated into the injection-moulded disposable chip. The core of the controlling instrument is a rotating heating bar construction providing fixed temperatures for fast cycling. PCR times of about half an hour (for 30 cycles) could be achieved for 120 µl reactions, making this system the fastest currently available high-volume PCR chip. The functionality of the system was shown by comparing automatically processed nasopharyngeal samples to ones processed manually according to the QIAGEN "ResPlex™ II Panel v2.0" respiratory virus detection kit. A prototype of the present instrument revealed slightly weaker signal intensities with a similar sensitivity in comparison to the commercially available kit and automated nucleic acid preparation devices, even without protocol optimization.

Automated microsystem for electrochemical detection of cancer markers.

The development of a fully automated microsystem housing an amperometric immunosensor is presented. The microfluidic cell integrates reagent storage and electrochemical immunodetection and was applied for the detection of breast cancer markers. The main advantage of this system is that no external fluidic storage is required and the instrumental setup is thus greatly simplified. The fluidics of the microsystem is computer controlled and requires minimal end-user intervention. The analytical performance of the device was compared with a manually driven system and applied for the amperometric detection of the carcinoembryonic antigen (CEA) and cancer antigen 15-3 (CA15-3). This automation methodology greatly improves the analytical performance of the immunosensor in terms of accuracy and reproducibility as evidenced by a reduction of LOD observed for CEA and CA15-3 with respect to a manually driven system. Finally, the automated microsystem was applied for the analysis of real patient serum samples, demonstrating excellent correlation with a commercial ELISA.

Integrated microfluidic platform for the electrochemical detection of breast cancer markers in patient serum samples.

A microsystem integrating electrochemical detection for the simultaneous detection of protein markers of breast cancer is reported. The microfluidic platform was realized by high precision milling of polycarbonate sheets and features two well distinguishable sections: a detection zone incorporating the electrode arrays and the fluid storage part. The detection area is divided into separate microfluidic chambers addressing selected electrodes for the measurement of samples and calibrators. The fluidic storage part of the platform consists of five reservoirs to store the reagents and sample, which are interfaced by septa. These reservoirs have the appropriate volume to run a single assay per cartridge and are manually filled. The liquids from the reservoirs are actuated by applying a positive air pressure (i.e.via a programmable syringe pump) through the septa and are driven to the detection zone via two turning valves. The application of the realised platform in the individual and simultaneous electrochemical detection of proteic cancer markers with very low detection limits are demonstrated. The microsystem has also been validated using real patient serum samples and excellent correlation with ELISA results obtained.

Compact, cost-efficient microfluidics-based stopped-flow device.

Stopped-flow technology is frequently used to monitor rapid (bio)chemical reactions with high temporal resolution, e.g., in dynamic investigations of enzyme reactions, protein interactions, or molecular transport mechanisms. However, conventional stopped-flow devices are often overly complex, voluminous, or costly. Moreover, excessive amounts of sample are often wasted owing to inefficient designs. To address these shortcomings, we propose a stopped-flow system based on microfluidic design principles. Our simple and cost-efficient approach offers distinct advantages over existing technology. In particular, the use of injection-molded disposable microfluidic chips minimizes required sample volumes and associated costs, simplifies handling, and prevents adverse cross-contamination effects. The cost of the system developed is reduced by an order of magnitude compared with the cost of commercial systems. The system contains a high-precision valve system for fluid control and features automated data acquisition capability with high temporal resolution. Analyses with two well-established reaction kinetics yielded a dead time of approximately 8-9 ms.

Amperometric immunosensor for carcinoembryonic antigen in colon cancer samples based on monolayers of dendritic bipodal scaffolds.

Detection of proteins that signal the presence or recurrence of cancer is a powerful therapeutic tool for effective early diagnosis and treatment. Carcinoembryonic antigen (CEA) has been extensively studied as a tumor marker in clinical diagnosis. We report on the development of an amperometric biosensor for the detection of CEA based on the immobilization of anti-CEA monoclonal antibody on a novel class of bipodal thiolated self-assembled monolayers containing reactive N-hydroxysuccinimide (NHS) ester end groups. The current variations showed a linear relationship with the concentration of CEA over the range of 0-200 ng/mL with a sensitivity of 3.8 nA x mL x ng(-1) and a detection limit of 0.2 ng/mL, which is well below the commonly accepted concentration threshold (5 ng/mL) used in clinical diagnosis. Real time and accelerated stability studies of the reporter antibody under various storage conditions demonstrated that the enzymatic activity and antibody affinity of the conjugate is retained for long periods of time in commercial stabilizing buffers such as StabilGuard Biomolecule Stabilizer, and a prediction of the stability trends was carried out using the kinetic and thermodynamic parameters obtained from the Arrhenius equation. The developed immunosensor as well as a commercially available enzyme-linked immunosorbent assay (ELISA) kit were successfully applied to the detection of CEA in serum samples obtained from colon cancer patients, and an excellent correlation of the levels of CEA measured was obtained. Ongoing work is looking at the incorporation of the developed biosensor into a platform for multiplexed simultaneous detection of several breast cancer related biomarkers.