Biosensors2015, 5(2), 199-222; doi:10.3390/bios5020199 - published 16 April 2015 Show/Hide Abstract
Abstract: The xCELLigence technology is a real-time cellular biosensor, which measures the net adhesion of cells to high-density gold electrode arrays printed on custom-designed E-plates. The strength of cellular adhesion is influenced by a myriad of factors that include cell type, cell viability, growth, migration, spreading and proliferation. We therefore hypothesised that xCELLigence biosensor technology would provide a valuable platform for the measurement of drug responses in a multitude of different experimental, clinical or pharmacological contexts. In this manuscript, we demonstrate how xCELLigence technology has been invaluable in the identification of (1) not only if cells respond to a particular drug, but (2) the window of drug responsiveness. The latter aspect is often left to educated guess work in classical end-point assays, whereas biosensor technology reveals the temporal profile of the response in real time, which enables both acute responses and longer term responses to be profiled within the same assay. In our experience, the xCELLigence biosensor technology is suitable for highly targeted drug assessment and also low to medium throughput drug screening, which produces high content temporal data in real time.
Biosensors2015, 5(2), 187-198; doi:10.3390/bios5020187 - published 13 April 2015 Show/Hide Abstract
Abstract: Despite the fact that the theoretical foundations of the sensitivity of waveguide grating based (bio)sensors are well-known, understood and their implications anticipated by the scientific community since several decades, to our knowledge, no prior publication has experimentally confirmed waveguide sensitivity for multiple film thicknesses, wavelengths and polarization of the propagating light. In this paper, the bulk refractive index sensitivity versus waveguide thickness of said refractometric sensors is experimentally determined and compared with predictions based on established theory. The effective refractive indices and the corresponding sensitivity were determined via the sensors’ coupling angles at different cover refractive indices for transverse electric as well as transverse magnetic polarized illumination at various wavelengths in the visible and near-infrared. The theoretical sensitivity was calculated by solving the mode equation for a three layer waveguide.
Biosensors2015, 5(2), 172-186; doi:10.3390/bios5020172 - published 2 April 2015 Show/Hide Abstract
Abstract: We present a novel tunnel magnetoresistance (TMR) scanning microscopeset-up capable of quantitatively imaging the magnetic stray field patterns of micron-sizedelements in 3D. By incorporating an Anderson loop measurement circuit for impedancematching, we are able to detect magnetoresistance changes of as little as 0.006%/Oe. By 3Drastering a mounted TMR sensor over our magnetic barcodes, we are able to characterisethe complex domain structures by displaying the real component, the amplitude and thephase of the sensor’s impedance. The modular design, incorporating a TMR sensor withan optical microscope, renders this set-up a versatile platform for studying and imagingimmobilised magnetic carriers and barcodes currently employed in biosensor platforms,magnetotactic bacteria and other complex magnetic domain structures of micron-sizedentities. The quantitative nature of the instrument and its ability to produce vector maps ofmagnetic stray fields has the potential to provide significant advantages over other commonlyused scanning magnetometry techniques.
Biosensors2015, 5(2), 158-171; doi:10.3390/bios5020158 - published 1 April 2015 Show/Hide Abstract
Abstract: We report use of U-shaped biconically tapered optical fibers (BTOF) as probes for label-free immunoassays. The tapered regions of the sensors were functionalized by immobilization of immunoglobulin-G (Ig-G) and tested for detection of anti-IgG at concentrations of 50 ng/mL to 50 µg/mL. Antibody-antigen reaction creates a biological nanolayer modifying the waveguide structure leading to a change in the sensor signal, which allows real-time monitoring. The kinetics of the antibody (mouse Ig-G)-antigen (rabbit anti-mouse IgG) reactions was studied. Hydrofluoric acid treatment makes the sensitive region thinner to enhance sensitivity, which we confirmed by experiments and simulations. The limit of detection for the sensor was estimated to be less than 50 ng/mL. Utilization of the rate of the sensor peak shift within the first few minutes of the antibody-antigen reaction is proposed as a rapid protein detection method.
Biosensors2015, 5(2), 141-157; doi:10.3390/bios5020141 - published 1 April 2015 Show/Hide Abstract
Abstract: A full label-free morphological and biochemical characterization is desirable to select spermatozoa during preparation for artificial insemination. In order to study these fundamental parameters, we take advantage of two attractive techniques: digital holography (DH) and Raman spectroscopy (RS). DH presents new opportunities for studying morphological aspect of cells and tissues non-invasively, quantitatively and without the need for staining or tagging, while RS is a very specific technique allowing the biochemical analysis of cellular components with a spatial resolution in the sub-micrometer range. In this paper, morphological and biochemical bovine sperm cell alterations were studied using these techniques. In addition, a complementary DH and RS study was performed to identify X- and Y-chromosome-bearing sperm cells. We demonstrate that the two techniques together are a powerful and highly efficient tool elucidating some important criterions for sperm morphological selection and sex-identification, overcoming many of the limitations associated with existing protocols.
Biosensors2015, 5(2), 131-140; doi:10.3390/bios5020131 - published 26 March 2015 Show/Hide Abstract
Abstract: An antibiotic rapamycin is one of the most commonly used immunosuppressive drugs, and also implicated for its anti-cancer activity. Hence, the determination of its blood level after organ transplantation or tumor treatment is of great concern in medicine. Although there are several rapamycin detection methods, many of them have limited sensitivity, and/or need complicated procedures and long assay time. As a novel fluorescent biosensor for rapamycin, here we propose “Q’-body”, which works on the fluorescence quench-release principle inspired by the antibody-based quenchbody (Q-body) technology. We constructed rapamycin Q’-bodies by linking the two interacting domains FKBP12 and FRB, whose association is triggered by rapamycin. The fusion proteins were each incorporated position-specifically with one of fluorescence dyes ATTO520, tetramethylrhodamine, or ATTO590 using a cell-free translation system. As a result, rapid rapamycin dose-dependent fluorescence increase derived of Q’-bodies was observed, especially for those with ATTO520 with a lowest detection limit of 0.65 nM, which indicates its utility as a novel fluorescent biosensor for rapamycin.