A platform of BRET-FRET hybrid biosensors for optogenetics, chemical screening, and in vivo imaging.

PubMed

Komatsu, Naoki; Terai, Kenta; Imanishi, Ayako; Kamioka, Yuji; Sumiyama, Kenta; Jin, Takashi; Okada, Yasushi; Nagai, Takeharu; Matsuda, Michiyuki

2018-06-12

Genetically encoded biosensors based on the principle of Förster resonance energy transfer comprise two major classes: biosensors based on fluorescence resonance energy transfer (FRET) and those based on bioluminescence energy transfer (BRET). The FRET biosensors visualize signaling-molecule activity in cells or tissues with high resolution. Meanwhile, due to the low background signal, the BRET biosensors are primarily used in drug screening. Here, we report a protocol to transform intramolecular FRET biosensors to BRET-FRET hybrid biosensors called hyBRET biosensors. The hyBRET biosensors retain all properties of the prototype FRET biosensors and also work as BRET biosensors with dynamic ranges comparable to the prototype FRET biosensors. The hyBRET biosensors are compatible with optogenetics, luminescence microplate reader assays, and non-invasive whole-body imaging of xenograft and transgenic mice. This simple protocol will expand the use of FRET biosensors and enable visualization of the multiscale dynamics of cell signaling in live animals.

Photonic crystal waveguide-based biosensor for detection of diseases

NASA Astrophysics Data System (ADS)

Chopra, Harshita; Kaler, Rajinder S.; Painam, Balveer

2016-07-01

A biosensor is a device that is used to detect the analytes or molecules of a sample by means of a binding mechanism. A two-dimensional photonic crystal waveguide-based biosensor is designed with a diamond-shaped ring resonator and two waveguides: a bus waveguide and a drop waveguide. The sensing mechanism is based on change in refractive index of the analytes, leading to a shift in the peak resonant wavelength. This mechanism can be used in the field of biomedical treatment where different body fluids such as blood, tears, saliva, or urine can be used as the analyte in which different components of the fluid can be detected. It can also be used to differentiate between the cell lines of a normal and an unhealthy human being. Average value of quality factor for this device comes out to be 1082.2063. For different analytes used, the device exhibits enhanced sensitivity and, hence, it is useful for the detection of diseases.

An immuno-biosensor system based on quartz crystal microbalance for avian influenza virus detection

NASA Astrophysics Data System (ADS)

Liu, Shengping; Chen, Guoming; Zhou, Qi; Wei, Yunlong

2007-12-01

For the quick detection of Avian Influenza Virus (AIV), a biosensor based on Quartz Crystal Microbalance (QCM) was fabricated according to the specific bonding principle between antibody and antigen. Staphylococcal Protein A (SPA) was extracted from Staphylococcus and purified. Then SPA was coated on the surface of QCM for immobilizing AIV monoclonal antibodies. The use of AIV monoclonal antibody could enhance the specificity of the immuno-biosensor. A multi-channel piezoelectricity detection system for the immuno-biosensor was developed. The system can work for the quick detection of AIV antigen in the case of the entirely aqueous status owe to one special oscillating circuit designed in this work. The optimum conditions of SPA coating and AIV monoclonal antibody immobilization were investigated utilizing the multi-channel detection system. The preliminary application of the immuno-biosensor system for detection of AIV was evaluated. Results indicate that the immuno-biosensor system can detect the AIV antigens with a linear range of 3-200ng/ml. The system can accomplish the detection of AIV antigens around 40 minutes.

Easter microplate dynamics

NASA Astrophysics Data System (ADS)

Neves, M. C.; Searle, R. C.; Bott, M. H. P.

2003-04-01

We use two-dimensional elastic finite element analysis, supplemented by strength estimates, to investigate the driving mechanism of the Easter microplate. Modeled stresses are compared with the stress indicators compiled from earthquake focal mechanisms and structural observations. The objective is to constrain the tectonic forces that govern the Easter microplate rotation and to test the microplate driving hypothesis proposed by [1993]. We infer that the mantle basal drag cannot drive the microplate rotation but opposes it, and that the asthenospheric viscosity is no more than about 1 × 1018 Pa s. At most, the basal drag comprises 20% of the force resisting microplate rotation. The outward pull of the main plates can drive the rotation by shear drag applied along the northern and southern boundaries of the microplate. However, we propose an additional driving force which arises from the strong variation of the ridge resistance force along the east and west rifts, so that the main driving torques come from the pull of the major plates acting across the narrowing and slowing rifts. This requires the strength to increase substantially toward the rift tips due to thickening of the brittle lithosphere as the spreading rate slows.

Detection of Myoglobin with an Open-Cavity-Based Label-Free Photonic Crystal Biosensor.

PubMed

Zhang, Bailin; Tamez-Vela, Juan Manuel; Solis, Steven; Bustamante, Gilbert; Peterson, Ralph; Rahman, Shafiqur; Morales, Andres; Tang, Liang; Ye, Jing Yong

2013-01-01

The label-free detection of one of the cardiac biomarkers, myoglobin, using a photonic-crystal-based biosensor in a total-internal-reflection configuration (PC-TIR) is presented in this paper. The PC-TIR sensor possesses a unique open optical microcavity that allows for several key advantages in biomolecular assays. In contrast to a conventional closed microcavity, the open configuration allows easy functionalization of the sensing surface for rapid biomolecular binding assays. Moreover, the properties of PC structures make it easy to be designed and engineered for operating at any optical wavelength. Through fine design of the photonic crystal structure, biochemical modification of the sensor surface, and integration with a microfluidic system, we have demonstrated that the detection sensitivity of the sensor for myoglobin has reached the clinically significant concentration range, enabling potential usage of this biosensor for diagnosis of acute myocardial infarction. The real-time response of the sensor to the myoglobin binding may potentially provide point-of-care monitoring of patients and treatment effects.

Distributed Feedback Laser Based on Single Crystal Perovskite

NASA Astrophysics Data System (ADS)

Sun, Shang; Xiao, Shumin; Song, Qinghai

2017-06-01

We demonstrate a single crystal perovskite based, with grating-structured photoresist on top, highly polarized distributed feedback laser. A lower laser threshold than the Fabry-Perot mode lasers from the same single crystal CH3NH3PbBr3 microplate was obtained. Single crystal CH3NH3PbBr3 microplates was synthesized with one-step solution processed precipitation method. Once the photoresist on top of the microplate was patterned with electron beam, the device was realized. This one-step fabrication process utilized the advantage of single crystal to the greatest extend. The ultra-low defect density in single crystalline microplate offer an opportunity for lower threshold lasing action compare with poly-crystal perovskite films. In the experiment, the lasing action based on the distributed feedback grating design was found with lower threshold and higher intensity than the Fabry-Perot mode lasers supported by the flat facets of the same microplate.

Design of highly sensitive multichannel bimetallic photonic crystal fiber biosensor

NASA Astrophysics Data System (ADS)

Hameed, Mohamed Farhat O.; Alrayk, Yassmin K. A.; Shaalan, Abdelhamid A.; El Deeb, Walid S.; Obayya, Salah S. A.

2016-10-01

A design of a highly sensitive multichannel biosensor based on photonic crystal fiber is proposed and analyzed. The suggested design has a silver layer as a plasmonic material coated by a gold layer to protect silver oxidation. The reported sensor is based on detection using the quasi transverse electric (TE) and quasi transverse magnetic (TM) modes, which offers the possibility of multichannel/multianalyte sensing. The numerical results are obtained using a finite element method with perfect matched layer boundary conditions. The sensor geometrical parameters are optimized to achieve high sensitivity for the two polarized modes. High-refractive index sensitivity of about 4750 nm/RIU (refractive index unit) and 4300 nm/RIU with corresponding resolutions of 2.1×10-5 RIU, and 2.33×10-5 RIU can be obtained according to the quasi TM and quasi TE modes of the proposed sensor, respectively. Further, the reported design can be used as a self-calibration biosensor within an unknown analyte refractive index ranging from 1.33 to 1.35 with high linearity and high accuracy. Moreover, the suggested biosensor has advantages in terms of compactness and better integration of microfluidics setup, waveguide, and metallic layers into a single structure.

Edge-driven microplate kinematics

USGS Publications Warehouse

Schouten, Hans; Klitgord, Kim D.; Gallo, David G.

1993-01-01

It is known from plate tectonic reconstructions that oceanic microplates undergo rapid rotation about a vertical axis and that the instantaneous rotation axes describing the microplate's motion relative to the bounding major plates are frequently located close to its margins with those plates, close to the tips of propagating rifts. We propose a class of edge-driven block models to illustrate how slip across the microplate margins, block rotation, and propagation of rifting may be related to the relative motion of the plates on either side. An important feature of these edge-driven models is that the instantaneous rotation axes are always located on the margins between block and two bounding plates. According to those models the pseudofaults or traces of disrupted seafloor resulting from the propagation of rifting between microplate and major plates may be used independently to approximately trace the continuous kinematic evolution of the microplate back in time. Pseudofault geometries and matching rotations of the Easter microplate show that for most of its 5 m.y. history, block rotation could be driven by the drag of the Nazca and Pacific plates on the microplate's edges rather than by a shear flow of mantle underneath.

Advantages and application of label-free detection assays in drug screening.

PubMed

Cunningham, Brian T; Laing, Lance G

2008-08-01

Adoption is accelerating for a new family of label-free optical biosensors incorporated into standard format microplates owing to their ability to enable highly sensitive detection of small molecules, proteins and cells for high-throughput drug discovery applications. Label-free approaches are displacing other detection technologies owing to their ability to provide simple assay procedures for hit finding/validation, accessing difficult target classes, screening the interaction of cells with drugs and analyzing the affinity of small molecule inhibitors to target proteins. This review describes several new drug discovery applications that are under development for microplate-based photonic crystal optical biosensors and the key issues that will drive adoption of the technology. Microplate-based optical biosensors are enabling a variety of cell-based assays, inhibition assays, protein-protein binding assays and protein-small molecule binding assays to be performed with high-throughput and high sensitivity.

Biochemical characterization and immobilization of Erwinia carotovoral-asparaginase in a microplate for high-throughput biosensing of l-asparagine.

PubMed

Labrou, Nikolaos E; Muharram, Magdy Mohamed

2016-10-01

l-Asparaginases (l-ASNase, E.C. 3.5.1.1) catalyze the conversion of l-asparagine to l-aspartic acid and ammonia. In the present work, a new form of l-ASNase from a strain of Erwinia carotovora (EcaL-ASNase) was cloned, expressed in Escherichia coli as a soluble protein and characterized. The enzyme was purified to homogeneity by a single-step procedure comprising ion-exchange chromatography. The properties of the recombinant enzyme were investigated employing kinetic analysis and molecular modelling and the kinetic parameters (Km, kcat) were determined for a number of substrates. The enzyme was used to assemble a microplate-based biosensor that was used for the development of a simple assay for the determination of l-asparagine in biological samples. In this sensor, the enzyme was immobilized by crosslinking with glutaraldehyde and deposited into the well of a microplate in 96-well format. The sensing scheme was based on the colorimetric measurement of ammonia formation using the Nessler's reagent. This format is ideal for micro-volume applications and allows the use of the proposed biosensor in high-throughput applications for monitoring l-asparagine levels in serum and foods samples. Calibration curve was obtained for l-asparagine, with useful concentration range 10-200μΜ. The biosensor had a detection limit of 10μM for l-asparagine. The method's reproducibility was in the order of ±3-6% and l-asparagine mean recoveries were 101.5%. Copyright © 2016 Elsevier Inc. All rights reserved.

Size-dependent phase transition in methylammonium lead iodide perovskite microplate crystals

PubMed Central

Li, Dehui; Wang, Gongming; Cheng, Hung-Chieh; Chen, Chih-Yen; Wu, Hao; Liu, Yuan; Huang, Yu; Duan, Xiangfeng

2016-01-01

Methylammonium lead iodide perovskite has attracted considerable recent interest for solution processable solar cells and other optoelectronic applications. The orthorhombic-to-tetragonal phase transition in perovskite can significantly alter its optical, electrical properties and impact the corresponding applications. Here, we report a systematic investigation of the size-dependent orthorhombic-to-tetragonal phase transition using a combined temperature-dependent optical, electrical transport and transmission electron microscopy study. Our studies of individual perovskite microplates with variable thicknesses demonstrate that the phase transition temperature decreases with reducing microplate thickness. The sudden decrease of mobility around phase transition temperature and the presence of hysteresis loops in the temperature-dependent mobility confirm that the orthorhombic-to-tetragonal phase transition is a first-order phase transition. Our findings offer significant fundamental insight on the temperature- and size-dependent structural, optical and charge transport properties of perovskite materials, and can greatly impact future exploration of novel electronic and optoelectronic devices from these materials. PMID:27098114

Size-dependent phase transition in methylammonium lead iodide perovskite microplate crystals

DOE PAGES

Li, Dehui; Wang, Gongming; Cheng, Hung -Chieh; ...

2016-04-21

Methylammonium lead iodide perovskite has attracted considerable recent interest for solution processable solar cells and other optoelectronic applications. The orthorhombic-to-tetragonal phase transition in perovskite can significantly alter its optical, electrical properties and impact the corresponding applications. Here, we report a systematic investigation of the size-dependent orthorhombic-to-tetragonal phase transition using a combined temperature-dependent optical, electrical transport and transmission electron microscopy study. Our studies of individual perovskite microplates with variable thicknesses demonstrate that the phase transition temperature decreases with reducing microplate thickness. The sudden decrease of mobility around phase transition temperature and the presence of hysteresis loops in the temperature-dependent mobility confirmmore » that the orthorhombic-to-tetragonal phase transition is a first-order phase transition. Lastly, our findings offer significant fundamental insight on the temperature-and size-dependent structural, optical and charge transport properties of perovskite materials, and can greatly impact future exploration of novel electronic and optoelectronic devices from these materials.« less

Quartz crystal microbalance (QCM) as biosensor for the detecting of Escherichia coli O157:H7

NASA Astrophysics Data System (ADS)

Thanh Ngo, Vo Ke; Giang Nguyen, Dang; Phuong Uyen Nguyen, Hoang; Tran, Van Man; Nguyen, Thi Khoa My; Phat Huynh, Trong; Lam, Quang Vinh; Dat Huynh, Thanh; Truong, Thi Ngoc Lien

2014-12-01

Although Escherichia coli (E. coli) is a commensalism organism in the intestine of humans and warm-blooded animals, it can be toxic at higher density and causes diseases, especially the highly toxic E. coli O157:H7. In this paper a quartz crystal microbalance (QCM) biosensor was developed for the detection of E. coli O157:H7 bacteria. The anti-E. coli O157:H7 antibodies were immobilized on a self-assembly monolayer (SAM) modified 5 MHz AT-cut quartz crystal resonator. The SAMs were activated with 16-mercaptopropanoic acid, in the presence of 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide (EDC) and ester N-hydroxysuccinimide (NHS). The result of changing frequency due to the adsorption of E. coli O157:H7 was measured by the QCM biosensor system designed and fabricated by ICDREC-VNUHCM. This system gave good results in the range of 102-107 CFU mL-1 E. coli O157:H7. The time of bacteria E. coli O157:H7 detection in the sample was about 50 m. Besides, QCM biosensor from SAM method was comparable to protein A method-based piezoelectric immunosensor in terms of the amount of immobilized antibodies and detection sensitivity.

Biosensor of endotoxin and sepsis

NASA Astrophysics Data System (ADS)

Shao, Yang; Wang, Xiang; Wu, Xi; Gao, Wei; He, Qing-hua; Cai, Shaoxi

2001-09-01

To investigate the relation between biosensor of endotoxin and endotoxin of plasma in sepsis. Method: biosensor of endotoxin was designed with technology of quartz crystal microbalance bioaffinity sensor ligand of endotoxin were immobilized by protein A conjugate. When a sample soliton of plasma containing endotoxin 0.01, 0.03, 0.06, 0.1, 0.5, 1.0Eu, treated with perchloric acid and injected into slot of quartz crystal surface respectively, the ligand was released from the surface of quartz crystal to form a more stable complex with endotoxin in solution. The endotoxin concentration corresponded to the weight change on the crystal surface, and caused change of frequency that occurred when desorbed. The result was biosensor of endotoxin might detect endotoxin of plasma in sepsis, measurements range between 0.05Eu and 0.5Eu in the stop flow mode, measurement range between 0.1Eu and 1Eu in the flow mode. The sensor of endotoxin could detect the endotoxin of plasm rapidly, and use for detection sepsis in clinically.

A semistatic microplate-based phytotoxicity test

DOE Office of Scientific and Technical Information (OSTI.GOV)

Radetski, C.M.; Ferard, J.F.; Blaise, C.

1995-02-01

A novel phytotoxicity test is described herein that employs a microplate equipped with membrane-bottomed wells. This MultiScreen[trademark] (Millipore Corp., Bedford, MA) microplate allows performance of a semistatic algal test, in which test medium is renewed periodically. With such a design, the algal test becomes comparable to other short-term tests used to evaluate chronic toxicity of chemicals and effluents. The EC50s obtained for Cu[sup 2+], Cd[sup 2+], Cr[sup 6+], atrazine, and one leachate sample (municipal sludge incinerator residue) with static and semistatic algal microplate tests were compared in this study. The semistatic microplate test revealed greater sensitivity than did the staticmore » microplate test.« less

Hydrogenated amorphous silicon nitride photonic crystals for improved-performance surface electromagnetic wave biosensors.

PubMed

Sinibaldi, Alberto; Descrovi, Emiliano; Giorgis, Fabrizio; Dominici, Lorenzo; Ballarini, Mirko; Mandracci, Pietro; Danz, Norbert; Michelotti, Francesco

2012-10-01

We exploit the properties of surface electromagnetic waves propagating at the surface of finite one dimensional photonic crystals to improve the performance of optical biosensors with respect to the standard surface plasmon resonance approach. We demonstrate that the hydrogenated amorphous silicon nitride technology is a versatile platform for fabricating one dimensional photonic crystals with any desirable design and operating in a wide wavelength range, from the visible to the near infrared. We prepared sensors based on photonic crystals sustaining either guided modes or surface electromagnetic waves, also known as Bloch surface waves. We carried out for the first time a direct experimental comparison of their sensitivity and figure of merit with surface plasmon polaritons on metal layers, by making use of a commercial surface plasmon resonance instrument that was slightly adapted for the experiments. Our measurements demonstrate that the Bloch surface waves on silicon nitride photonic crystals outperform surface plasmon polaritons by a factor 1.3 in terms of figure of merit.

Scribed transparency microplates mounted on a modified standard microplate.

PubMed

Cheong, Brandon Huey-Ping; Chua, Wei Seong; Liew, Oi Wah; Ng, Tuck Wah

2014-08-01

The immense cost effectiveness of using transparencies as analyte handling implements in microplate instrumentation offers the possibility of application even in resource-limited laboratories. In this work, a standard microplate was adapted to serve as the permanent base for disposable scribed transparencies. The approach is shown to ameliorate evaporation, which can affect assay accuracy when analytes need to be incubated for some time. It also offers assurance against fluorescence measurement errors due to the cross-talk of samples from adjacent wells. Copyright © 2014 Elsevier Inc. All rights reserved.

The Galapagos Microplate Revealed

NASA Astrophysics Data System (ADS)

Smith, D. K.; Schouten, H.; Cann, J. R.; Zhu, W.; Montesi, L. G.; Mitchell, G. A.

2009-12-01

We report a new bathymetry survey of the Galapagos microplate (GMP), which separates the Pacific, Nazca, and Cocos plates at the Galapagos Triple Junction. Prior to the formation of the microplate, 1.5-1.0 Ma, there was a succession of transient minor rifts forming triple junctions north and south of the propagating Cocos-Nazca rift (see Schouten et al. abstract). As proposed by Lonsdale (1988) the formation of a large near-axis seamount coincided with the initiation of the GMP and stabilized rifting on its southern boundary, now called Dietz Deep Rift. Lonsdale also proposed that the GMP was rotating clockwise at 6 degrees/my. Schouten et al. (1993) and Klein et al. (2005) applied an edge-driven microplate model to the GMP to understand its kinematics and predicted rotation rates of 30-40 degrees/my and 22 degrees/my, respectively. These interpretations and predictions were based on sparse bathymetry data. In early 2009 (AT 15-41), we mapped the Galapagos microplate in its entirety to understand more fully the conditions that led to the stabilization of the southern triple junction at Dietz Deep Rift and to constrain the rotation rate of the microplate. Our new data show the two highly contrasted sections of Dietz Deep Rift. The northeastern section contains Dietz Deep, a 2 km deep basin, within a fault-dominated rift valley about 20 km wide; subsidiary rifts occur to the south. Sidescan data indicate that extension in this broadly rifted area has been largely amagmatic. The southwestern section of Dietz Deep Rift is dominated by a variety of volcanic constructions in which faulting plays a minor part. The volcanism has resulted in two large seamounts and a number of volcanic ridges running parallel to the fault dominated rift valley. The largest volcanic ridge is steep-sided and straight, and extends to intersect the East Pacific Rise (EPR) at 1 10’N to form the triple junction. Other minor volcanic ridges occur in the SW section of the microplate fanning

A novel assay for detecting canine parvovirus using a quartz crystal microbalance biosensor.

PubMed

Kim, Yong Kwan; Lim, Seong-In; Choi, Sarah; Cho, In-Soo; Park, Eun-Hye; An, Dong-Jun

2015-07-01

Rapid and accurate diagnosis is crucial to reduce both the shedding and clinical signs of canine parvovirus (CPV). The quartz crystal microbalance (QCM) is a new tool for measuring frequency changes associated with antigen-antibody interactions. In this study, the QCM biosensor and ProLinker™ B were used to rapidly diagnosis CPV infection. ProLinker™ B enables antibodies to be attached to a gold-coated quartz surface in a regular pattern and in the correct orientation for antigen binding. Receiver operating characteristics (ROC) curves were used to set a cut-off value using reference CPVs (two groups: one CPV-positive and one CPV-negative). The ROC curves overlapped and the point of intersection was used as the cut-off value. A QCM biosensor with a cut-off value of -205 Hz showed 95.4% (104/109) sensitivity and 98.0% (149/152) specificity when used to test 261 field fecal samples compared to PCR. In conclusion, the QCM biosensor described herein is eminently suitable for the rapid diagnosis of CPV infection with high sensitivity and specificity. Therefore, it is a promising analytical tool that will be useful for clinical diagnosis, which requires rapid and reliable analyses. Copyright © 2015 Elsevier B.V. All rights reserved.

Biosensors for hepatitis B virus detection.

PubMed

Yao, Chun-Yan; Fu, Wei-Ling

2014-09-21

A biosensor is an analytical device used for the detection of analytes, which combines a biological component with a physicochemical detector. Recently, an increasing number of biosensors have been used in clinical research, for example, the blood glucose biosensor. This review focuses on the current state of biosensor research with respect to efficient, specific and rapid detection of hepatitis B virus (HBV). The biosensors developed based on different techniques, including optical methods (e.g., surface plasmon resonance), acoustic wave technologies (e.g., quartz crystal microbalance), electrochemistry (amperometry, voltammetry and impedance) and novel nanotechnology, are also discussed.

Study and development of label-free optical biosensors for biomedical applications

NASA Astrophysics Data System (ADS)

Choi, Charles J.

For the majority of assays currently performed, fluorescent or colorimetric chemical labels are commonly attached to the molecules under study so that they may be readily visualized. The methods of using labels to track biomolecular binding events are very sensitive and effective, and are employed as standardized assay protocol across research labs worldwide. However, using labels induces experimental uncertainties due to the effect of the label on molecular conformation, active binding sites, or inability to find an appropriate label that functions equivalently for all molecules in an experiment. Therefore, the ability to perform highly sensitive biochemical detection without the use of fluorescent labels would further simplify assay protocols and would provide quantitative kinetic data, while removing experimental artifacts from fluorescent quenching, shelf-life, and background fluorescence phenomena. In view of the advantages mentioned above, the study and development of optical label-free sensor technologies have been undertaken here. In general, label-free photonic crystal (PC) biosensors and metal nanodome array surface-enhanced Raman scattering (SERS) substrates, both of which are fabricated by nanoreplica molding process, have been used as the method to attack the problem. Chapter 1 shows the work on PC label-free biosensor incorporated microfluidic network for bioassay performance enhancement and kinetic reaction rate constant determination. Chapter 2 describes the work on theoretical and experimental comparison of label-free biosensing in microplate, microfluidic, and spot-based affinity capture assays. Chapter 3 shows the work on integration of PC biosensor with actuate-to-open valve microfluidic chip for pL-volume combinatorial mixing and screening application. In Chapter 4, the development and characterization of SERS nanodome array is shown. Lastly, Chapter 5 describes SERS nanodome sensor incorporated tubing for point-of-care monitoring of

Microplates with adaptive surfaces.

PubMed

Akbulut, Meshude; Lakshmi, Dhana; Whitcombe, Michael J; Piletska, Elena V; Chianella, Iva; Güven, Olgun; Piletsky, Sergey A

2011-11-14

Here we present a new and versatile method for the modification of the well surfaces of polystyrene microtiter plates (microplates) with poly(N-phenylethylene diamine methacrylamide), (poly-NPEDMA). The chemical grafting of poly-NPEDMA to the surface of microplates resulted in the formation of thin layers of a polyaniline derivative bearing pendant methacrylamide double bonds. These were used as the attachment point for various functional polymers through photochemical grafting of various, for example, acrylate and methacrylate, polymers with different functionalities. In a model experiment, we have modified poly-NPEDMA-coated microplates with a small library of polymers containing different functional groups using a two-step approach. In the first step, double bonds were activated by UV irradiation in the presence of N,N-diethyldithiocarbamic acid benzyl ester (iniferter). This enabled grafting of the polymer library in the second step by UV irradiation of solutions of the corresponding monomers in the microplate wells. The uniformity of coatings was confirmed spectrophotometrically, by microscopic imaging and by contact angle measurements (CA). The feasibility of the current technology has been shown by the generation of a small library of polymers grafted to the microplate well surfaces and screening of their affinity to small molecules, such as atrazine, a trio of organic dyes, and a model protein, bovine serum albumin (BSA). The stability of the polymers, reproducibility of measurement, ease of preparation, and cost-effectiveness make this approach suitable for applications in high-throughput screening in the area of materials research.

Liquid crystal based biosensors for bile acid detection

NASA Astrophysics Data System (ADS)

He, Sihui; Liang, Wenlang; Tanner, Colleen; Fang, Jiyu; Wu, Shin-Tson

2013-03-01

The concentration level of bile acids is a useful indicator for early diagnosis of liver diseases. The prevalent measurement method in detecting bile acids is the chromatography coupled with mass spectrometry, which is precise yet expensive. Here we present a biosensor platform based on liquid crystal (LC) films for the detection of cholic acid (CA). This platform has the advantage of low cost, label-free, solution phase detection and simple analysis. In this platform, LC film of 4-Cyano-4'-pentylbiphenyl (5CB) was hosted by a copper grid supported with a polyimide-coated glass substrate. By immersing into sodium dodecyl sulfate (SDS) solution, the LC film was coated with SDS which induced a homeotropic anchoring of 5CB. Addition of CA introduced competitive adsorption between CA and SDS at the interface, triggering a transition from homeotropic to homogeneous anchoring. The detection limit can be tuned by changing the pH value of the solution from 12uM to 170uM.

Counter-rotating microplates at the Galapagos triple junction.

PubMed

Klein, Emily M; Smith, Deborah K; Williams, Clare M; Schouten, Hans

2005-02-24

An 'incipient' spreading centre east of (and orthogonal to) the East Pacific Rise at 2 degrees 40' N has been identified as forming a portion of the northern boundary of the Galapagos microplate. This spreading centre was described as a slowly diverging, westward propagating rift, tapering towards the East Pacific Rise. Here we present evidence that the 'incipient rift' has also rifted towards the east and opens anticlockwise about a pivot at its eastern end. The 'incipient rift' then bounds a second microplate, north of the clockwise-rotating Galapagos microplate. The Galapagos triple junction region, in the eastern equatorial Pacific Ocean, thus consists of two counter-rotating microplates partly separated by the Hess Deep rift. Our kinematic solution for microplate motion relative to the major plates indicates that the two counter-rotating microplates may be treated as rigid blocks driven by drag on the microplates' edges3.

Kinematics of the Danakil microplate

NASA Astrophysics Data System (ADS)

Eagles, Graeme; Gloaguen, Richard; Ebinger, Cynthia

2002-10-01

A refinement and extrapolation of recent motion estimates for the Danakil microplate, based on ancient kinematic indicators in the Afar region, describes the evolution of a microplate in the continental realm. The Danakil horst is an elevated part of this microplate, exposing a Precambrian basement within the Afar depression, the site of the Nubia-Somalia-Arabia triple junction. We compare evidence for strike- or oblique-slip faults in data from the Afar depression and southern Red Sea to small circles about published poles of rotation for the Danakil microplate with respect to Nubia. A reconstruction about the preferred pole reunites lengths of a Precambrian shear zone on the Nubia and Danakil sides and preserves a uniform basement fabric strike through Nubia, Danakil and Yemen. Since at least magnetic chron C5 (˜11 Ma) Danakil rotated about a different pole with respect to Nubia than either Somalia or Arabia, but between chrons C5 and C2A Nubia-Danakil motion was a close approximation to Nubia-Somalia motion. Since C2A relative motions of the Danakil microplate have been independent of movements on any of the neighbouring plate boundaries. We relate this to the onset of oceanic-type accretion within Afar. The resulting eastwards acceleration of Danakil was accommodated by westwards propagation of the Gulf of Aden rift that became the new, discrete, plate boundary between the Danakil microplate and the Somalia plate. Present-day activity suggests that the Red Sea and Aden rifts will link through Afar, thereby isolating the Danakil horst as a microcontinent on the Arabian margin.

Novel multichannel surface plasmon resonance photonic crystal fiber biosensor

NASA Astrophysics Data System (ADS)

Hameed, Mohamed Farhat O.; Alrayk, Yassmin K. A.; Shaalan, A. A.; El Deeb, Walid S.; Obayya, S. S. A.

2016-04-01

In this paper, a novel design of highly sensitive biosensor based on photonic crystal fiber is presented and analyzed using full vectorial finite element method. The suggested design depends on using silver layer as a plasmonic active material coated by a gold layer to protect silver oxidation. The reported sensor is based on the detection using the quasi transverse electric (TE) and quasi transverse magnetic (TM) modes which offers the possibility of multi-channel/multi-analyte sensing. The sensor geometrical parameters are optimized to achieve high sensitivity for the two polarized modes. High refractive index sensitivity of about 4750 nm/RIU (refractive index unit) and 4300 nm/RIU with corresponding resolutions of 2.1×10-5 RIU, and 2.33×10-5 RIU can be obtained for the quasi TM and quasi TE modes, respectively.

Silicon on-chip bandpass filters for the multiplexing of high sensitivity photonic crystal microcavity biosensors

PubMed Central

Chakravarty, Swapnajit; Yang, Chun-Ju; Wang, Zheng; Tang, Naimei; Fan, Donglei; Chen, Ray T.

2015-01-01

A method for the dense integration of high sensitivity photonic crystal (PC) waveguide based biosensors is proposed and experimentally demonstrated on a silicon platform. By connecting an additional PC waveguide filter to a PC microcavity sensor in series, a transmission passband is created, containing the resonances of the PC microcavity for sensing purpose. With proper engineering of the passband, multiple high sensitivity PC microcavity sensors can be integrated into microarrays and be interrogated simultaneously between a single input and a single output port. The concept was demonstrated with a 2-channel L55 PC biosensor array containing PC waveguide filters. The experiment showed that the sensors on both channels can be monitored simultaneously from a single output spectrum. Less than 3 dB extra loss for the additional PC waveguide filter is observed. PMID:25829549

Silicon on-chip bandpass filters for the multiplexing of high sensitivity photonic crystal microcavity biosensors

DOE Office of Scientific and Technical Information (OSTI.GOV)

Yan, Hai, E-mail: hai.yan@utexas.edu; Zou, Yi; Yang, Chun-Ju

A method for the dense integration of high sensitivity photonic crystal (PC) waveguide based biosensors is proposed and experimentally demonstrated on a silicon platform. By connecting an additional PC waveguide filter to a PC microcavity sensor in series, a transmission passband is created, containing the resonances of the PC microcavity for sensing purpose. With proper engineering of the passband, multiple high sensitivity PC microcavity sensors can be integrated into microarrays and be interrogated simultaneously between a single input and a single output port. The concept was demonstrated with a 2-channel L55 PC biosensor array containing PC waveguide filters. The experimentmore » showed that the sensors on both channels can be monitored simultaneously from a single output spectrum. Less than 3 dB extra loss for the additional PC waveguide filter is observed.« less

Chitosan-coated polystyrene microplate for covalent immobilization of enzyme.

PubMed

Zhang, Yaodong; Li, Li; Yu, Caihong; Hei, Tingting

2011-10-01

Microplates made of polystyrene have been widely used for immunoassays. Protein molecules that have been immobilized on a hydrophobic polystyrene microplate by passive adsorption lose their activity and suffer considerable denaturation. A new chitosan-coated microplate suitable for the covalent immobilization of enzymes has been developed. The primary amino groups of chitosan were exploited for this covalent coupling of proteins. The optical transmittance of the chitosan-coated microplate, at wavelengths of 400-800 nm, was estimated to be suitable for its application in chromogenic reaction-based bioassays. The immobilization efficiency of the chitosan-coated microplate was demonstrated to be far superior to that of a conventional microplate when tested using acetylcholinesterase (AChE) and β-glucosidase as model biomolecules, and the chitosan-coated microplate may thus have potential applications in biosensing and bioreactor systems. © Springer-Verlag 2011

Low-cost and highly efficient DNA biosensor for heavy metal ion using specific DNAzyme-modified microplate and portable glucometer-based detection mode.

PubMed

Zhang, Jin; Tang, Ying; Teng, Liumei; Lu, Minghua; Tang, Dianping

2015-06-15

A simple and low-cost DNA sensing platform based on Pb(2+)-specific DNAzyme-modified microplate was successfully developed for highly sensitive monitoring of lead ion (Pb(2+), one kind of toxic heavy metal ion) in the environmental samples coupling with a portable personal glucometer (PGM)-based detection mode. The detection cell was first prepared simply by means of immobilizing the DNAzyme on the streptavidin-modified microplate. Gold nanoparticle labeled with single-stranded DNA and invertase (Enz-AuNP-DNA) was utilized as the signal-transduction tag to produce PGM substrate (glucose). Upon addition of lead ion into the microplate, the substrate strand of the immobilized DNAzyme was catalytically cleaved by target Pb(2+), and the newly generated single-strand DNA in the microplate could hybridize again with the single-stranded DNA on the Enz-AuNP-DNA. Accompanying with the Enz-AuNP-DNA, the carried invertase could convert sucrose into glucose. The as-produced glucose could be monitored by using a widely accessible PGM for in situ amplified digital readout. Based on Enz-AuNP-DNA amplification strategy, as low as 1.0 pM Pb(2+) could be detected under the optimal conditions. Moreover, the methodology also showed good reproducibility and high selectivity toward target Pb(2+) against other metal ions because of highly specific Pb(2+)-dependent DNAzyme, and was applicable for monitoring Pb(2+) in the naturally contaminated sewage and spiked drinking water samples. Copyright © 2015 Elsevier B.V. All rights reserved.

Evaluation of a novel label-free photonic-crystal biosensor imaging system for the detection of prostate cancer cells

NASA Astrophysics Data System (ADS)

DeLuna, Frank; Ding, XiaoFie; Sun, Lu-Zhe; Ye, Jing Yong

2017-02-01

Biomarker screening for prostate-specific antigen (PSA) is the current clinical standard for detection of prostate cancer. However this method has shown many limitations, mainly in its specificity, which can lead to a high false positive rate. Thus, there is a growing need in developing a more specific detection system for prostate cancer. Using a Photonic- Crystal-based biosensor in a Total-Internal-Reflection (PC-TIR) configuration, we demonstrate the use of refractive index (RI) to accomplish label-free detection of prostate cancer cells against non-cancerous prostate epithelial cells. The PC-TIR biosensor possesses an open microcavity, which in contrast to traditional closed microcavities, allows for easier access of analyte molecules or cells to interact with its sensing surface. In this study, an imaging system was designed using the PC-TIR biosensor to quantify cell RI as the contrast parameter for prostate cancer detection. Non-cancerous BPH-1 prostate epithelial cells and prostate cancer PC-3 cells were placed on a single biosensor and measured concurrently. Recorded image data was then analyzed through a home-built MatLab program. Results demonstrate that RI is a suitable variable for differentiation between prostate cancer cells and non-cancerous prostate epithelial cells. Our study shows clinical potential in utilizing RI test for the detection of prostate cancer.

170-MHz electrodeless quartz crystal microbalance biosensor: capability and limitation of higher frequency measurement.

PubMed

Ogi, Hirotsugu; Nagai, Hironao; Naga, Hironao; Fukunishi, Yuji; Hirao, Masahiko; Nishiyama, Masayoshi

2009-10-01

We develop a highly sensitive quartz crystal microbalance (QCM) biosensor with a fundamental resonance frequency of 170 MHz. A naked AT-cut quartz plate of 9.7 microm thick is set in a sensor cell. Its shear vibration is excited by the line wire, and the vibration signals are detected by the other line wire, achieving the noncontacting measurement of the resonance frequency. The mass sensitivity of the 170 MHz QCM biosensor is 15 pg/(cm2 Hz), which is better than that of a conventional 5 MHz QCM by 3 orders of magnitude. Its high sensitivity is confirmed by detecting human immunoglobulin G (hIgG) via Staphylococcus protein A immobilized nonspecifically on both surfaces of the quartz plate. The detection limit is 0.5 pM. Limitation of the high-frequency QCM measurement is then theoretically discussed with a continuum mechanics model for a plate with point masses connected by elastic springs. The result indicates that a QCM measurement will break down at frequencies one-order-of-magnitude higher than the local resonance frequency at specific binding cites.

Three-dimensionally ordered macroporous (3DOM) gold-nanoparticle-doped titanium dioxide (GTD) photonic crystals modified electrodes for hydrogen peroxide biosensor.

PubMed

Li, Jianlin; Han, Tao; Wei, Nannan; Du, Jiangyan; Zhao, Xiangwei

2009-12-15

Gold nanoparticles have been introduced into the wall framework of titanium dioxide photonic crystals by the colloidal crystal template technique. The three-dimensionally ordered macroporous gold-nanoparticle-doped titanium dioxide (3DOM GTD) film was modified on the indium-tin oxide (ITO) electrode surface and used for the hydrogen peroxide biosensor. The direct electron transfer and electrocatalysis of horseradish peroxidase (HRP) immobilized on this film have been investigated. The 3DOM GTD film could provide a good microenvironment for retaining the biological bioactivity, large internal area, and superior conductivity. The HRP/3DOM GTD/ITO electrode exhibited two couples of redox peaks corresponding to the HRP intercalated in the mesopores and adsorbed on the external surface of the film with the formal potential of -0.19 and -0.52V in 0.1M PBS (pH 7.4), respectively. The HRP intercalated in the mesopores showed a surface-controlled process with a single proton transfer. The direct electron transfer between the adsorbed HRP and the electrode is achieved without the aid of an electron mediator. The H(2)O(2) biosensor displayed a rapid eletrocatalytic response (less than 3s), a wide linear range from 0.5 microM to 1.4mM with a detection limit of 0.2 microM, high sensitivity (179.9 microAmM(-1)), good stability and reproducibility. Compared with the free-Au doped titanium dioxide photonic crystals modified electrode, the GTD modified electrode could greatly enhance the response current signal, linear detection range and higher sensitivity. The 3DOM GTD provided a new matrix for protein immobilization and direct transfer study and opened a way for low conductivity electrode biosensor.

Plasma-Treated Microplates with Enhanced Protein Recoveries and Minimized Extractables

PubMed Central

Weikart, Christopher M.; Klibanov, Alexander M.; Breeland, Adam P.; Taha, Ahmad H.; Maurer, Brian R.; Martin, Steven P.

2016-01-01

SiO2 Medical Products, Inc. (SiO) has developed a proprietary technology that greatly enhances protein recoveries and reduces extractables from commercial microplates used for bioanalytical assays and storage of biologics. SiO technology is based on plasma treatment that chemically modifies the surface of polypropylene with predominantly hydrogen-bond-acceptor uncharged polar groups. The resultant surface resists nonspecific protein adsorption over a wide range of protein concentrations, thereby eliminating the need to passivate (and hence potentially contaminate) the microplates with blocking proteins. High shelf-life stability and cleanliness of the plasma-treated microplates have been demonstrated using five different proteins for two common microplate formats. The protein recovery performance of plasma-treated microplates is found to be higher compared with commercial low-protein-binding microplates. PMID:27651466

India-Eurasia collision triggers formation of an oceanic microplate

NASA Astrophysics Data System (ADS)

Matthews, Kara; Müller, Dietmar; Sandwell, David

2016-04-01

Detailed mapping of seafloor tectonic fabric in the Indian Ocean, using high-resolution satellite-derived vertical gravity gradient data, reveals an extinct Pacific-style oceanic microplate - the Mammerickx Microplate - west of the Ninetyeast Ridge. It is one of the first Pacific-style microplates to be mapped outside the Pacific basin, suggesting that geophysical conditions during formation probably resembled those that have dominated at eastern Pacific ridges. The microplate formed at the Indian-Antarctic ridge and is bordered by an extinct ridge in the north and pseudofault in the south, whose conjugate is located north of the Kerguelen Plateau. Independent microplate rotation is indicated by asymmetric pseudofaults and rotated abyssal hill fabric, also identified in multibeam data. Magnetic anomaly picks and age estimates calculated from published spreading rates suggest formation during chron 21o (~47.3 Ma). Plate reorganizations can trigger ridge propagation and microplate development, and we propose that formation of the Mammerickx Microplate is linked with the initial 'soft' stage of the India-Eurasia collision. The collision altered the stress regime at the Indian-Antarctic ridge, leading to a change in segmentation and ridge propagation from an establishing transform fault. Fast Indian-Antarctic spreading that preceded microplate formation, and Kerguelen Plume activity may have facilitated ridge propagation via the production of thin and weak lithosphere. However, both factors had been present for tens of millions of years and are therefore unlikely to have triggered the event. Prior to the collision, this combination of fast spreading and plume activity was responsible for the production of a wide region of undulate seafloor to the north of the extinct ridge and 'W' shaped lineations that record back and forth ridge propagation. Microplate formation provides a means of dating the onset of the India-Eurasia collision, and is completely independent of and

A highly-sensitive label-free biosensor based on two dimensional photonic crystals with negative refraction

NASA Astrophysics Data System (ADS)

Malmir, Narges; Fasihi, Kiazand

2017-11-01

In this work, we present a novel high-sensitive optical label-free biosensor based on a two-dimensional photonic crystal (2D PC). The suggested structure is composed of a negative refraction structure in a hexagonal lattice PC, along with a positive refraction structure which is arranged in a square lattice PC. The frequency shift of the transmission peak is measured respect to the changes of refractive indices of the studied materials (the blood plasma, water, dry air and normal air). The studied materials are filled into a W1 line-defect waveguide which is located in the PC structure with positive refraction (the microfluidic nanochannel). Our numerical simulations, which are based on finite-difference time-domain (FDTD) method, show that in the proposed structure, a sensitivity about 1100 nm/RIU and a transmission efficiency more than 75% can be achieved. With this design, to the best of our knowledge, the obtained sensitivity and the transmission efficiency are one of the highest values in the reported PC label-free biosensors.

Laryngotracheal reconstruction with resorbable microplate buttressing.

PubMed

Javia, Luv R; Zur, Karen B

2012-04-01

In patients undergoing laryngotracheal reconstruction (LTR), malacic segments of trachea can pose challenges to successful reconstruction. Malacic segments may inadequately support cartilage grafts used in augmentation surgery, sometimes requiring cricotracheal or tracheal resections. We describe a novel technique of LTR with resorbable microplate buttressing of malacic lateral tracheal segments. Retrospective case series. Review of technique, treatment outcomes, and complications of seven children with subglottic stenosis and tracheomalacia requiring a microplate-augmented LTR technique. Seven infants ranging from 26 months to 9 years of age successfully underwent LTR for subglottic stenosis. Six children had a grade III subglottic stenosis. The seventh child had grade II subglottic stenosis, bilateral vocal fold paralysis, an elliptical cricoid, and an obstructing giant suprastomal fibroma. Five children underwent a double-stage LTR with resorbable microplates sutured bilaterally to support severely malacic lateral tracheal segments. A cricotracheal resection would not have been feasible in one child due to the resection length and inadequate tracheal mobilization. Two children underwent a single-stage LTR with unilateral application of a microplate. Six children were decannulated within 3 months and continue without airway symptoms or complications. One child, who is just over 2 months from reconstructive surgery, is being setup for decannulation. No complications were encountered. LTR with resorbable microplate buttressing of malacic lateral tracheal segments is technically feasible, safe, and can avoid more extensive surgery requiring tracheal resection. Further experience may support the use of this technique in challenging airway reconstructions. Copyright © 2012 The American Laryngological, Rhinological, and Otological Society, Inc.

High-throughput living cell-based optical biosensor for detection of bacterial lipopolysaccharide (LPS) using a red fluorescent protein reporter system.

PubMed

Jiang, Hui; Jiang, Donglei; Shao, Jingdong; Sun, Xiulan; Wang, Jiasheng

2016-11-14

Due to the high toxicity of bacterial lipopolysaccharide (LPS), resulting in sepsis and septic shock, two major causes of death worldwide, significant effort is directed toward the development of specific trace-level LPS detection systems. Here, we report sensitive, user-friendly, high-throughput LPS detection in a 96-well microplate using a transcriptional biosensor system, based on 293/hTLR4A-MD2-CD14 cells that are transformed by a red fluorescent protein (mCherry) gene under the transcriptional control of an NF-κB response element. The recognition of LPS activates the biosensor cell, TLR4, and the co-receptor-induced NF-κB signaling pathway, which results in the expression of mCherry fluorescent protein. The novel cell-based biosensor detects LPS with specificity at low concentration. The cell-based biosensor was evaluated by testing LPS isolated from 14 bacteria. Of the tested bacteria, 13 isolated Enterobacteraceous LPSs with hexa-acylated structures were found to increase red fluorescence and one penta-acylated LPS from Pseudomonadaceae appeared less potent. The proposed biosensor has potential for use in the LPS detection in foodstuff and biological products, as well as bacteria identification, assisting the control of foodborne diseases.

Guidelines for Microplate Selection in High Content Imaging.

PubMed

Trask, Oscar J

2018-01-01

Since the inception of commercialized automated high content screening (HCS) imaging devices in the mid to late 1990s, the adoption of media vessels typically used to house and contain biological specimens for interrogation has transitioned from microscope slides and petri dishes into multi-well microtiter plates called microplates. The early 96- and 384-well microplates commonly used in other high-throughput screening (HTS) technology applications were often not designed for optical imaging. Since then, modifications and the use of next-generation materials with improved optical clarity have enhanced the quality of captured images, reduced autofocusing failures, and empowered the use of higher power magnification objectives to resolve fine detailed measurements at the subcellular pixel level. The plethora of microplates and their applications requires practitioners of high content imaging (HCI) to be especially diligent in the selection and adoption of the best plates for running longitudinal studies or larger screening campaigns. While the highest priority in experimental design is the selection of the biological model, the choice of microplate can alter the biological response and ultimately may change the experimental outcome. This chapter will provide readers with background, troubleshooting guidelines, and considerations for choosing an appropriate microplate.

Overview of Piezoelectric Biosensors, Immunosensors and DNA Sensors and Their Applications.

PubMed

Pohanka, Miroslav

2018-03-19

Piezoelectric biosensors are a group of analytical devices working on a principle of affinity interaction recording. A piezoelectric platform or piezoelectric crystal is a sensor part working on the principle of oscillations change due to a mass bound on the piezoelectric crystal surface. In this review, biosensors having their surface modified with an antibody or antigen, with a molecularly imprinted polymer, with genetic information like single stranded DNA, and biosensors with bound receptors of organic of biochemical origin, are presented and discussed. The mentioned recognition parts are frequently combined with use of nanoparticles and applications in this way are also introduced. An overview of the current literature is given and the methods presented are commented upon.

Microplates in liquid chromatography--new solution in clinical research? A review.

PubMed

Krcmova, Lenka; Solichova, Dagmar; Solich, Petr

2013-10-15

Microplates are routinely used in Radio- or Immuno-assays. Recently, microplates have found use not only in analytical but also in the pre-analytical phase in bioanalyses (sample storage, sample preparation). New connection of this technology to liquid chromatography could be economical, fast and simple solution for many routine laboratories handling large sequences of biological samples. This review summarises the application of microplates in bioanalytical laboratories. Different types of sorbents, materials and shapes of microplates are discussed, and the main advantages and disadvantages of microplates used in clinical research are presented. Copyright © 2013 Elsevier B.V. All rights reserved.

Oceanic microplate formation records the onset of India-Eurasia collision

NASA Astrophysics Data System (ADS)

Matthews, Kara J.; Dietmar Müller, R.; Sandwell, David T.

2016-01-01

Mapping of seafloor tectonic fabric in the Indian Ocean, using high-resolution satellite-derived vertical gravity gradient data, reveals an extinct Pacific-style oceanic microplate ('Mammerickx Microplate') west of the Ninetyeast Ridge. It is one of the first Pacific-style microplates to be mapped outside the Pacific basin, suggesting that geophysical conditions during formation probably resembled those that have dominated at eastern Pacific ridges. The microplate formed at the Indian-Antarctic ridge and is bordered by an extinct ridge in the north and pseudofault in the south, whose conjugate is located north of the Kerguelen Plateau. Independent microplate rotation is indicated by asymmetric pseudofaults and rotated abyssal hill fabric, also seen in multibeam data. Magnetic anomaly picks and age estimates calculated from published spreading rates suggest formation during chron 21o (∼47.3 Ma). Plate reorganizations can trigger ridge propagation and microplate development, and we propose that Mammerickx Microplate formation is linked with the India-Eurasia collision (initial 'soft' collision). The collision altered the stress regime at the Indian-Antarctic ridge, leading to a change in segmentation and ridge propagation from an establishing transform. Fast Indian-Antarctic spreading that preceded microplate formation, and Kerguelen Plume activity, may have facilitated ridge propagation via the production of thin and weak lithosphere; however both factors had been present for tens of millions of years and are therefore unlikely to have triggered the event. Prior to the collision, the combination of fast spreading and plume activity was responsible for the production of a wide region of undulate seafloor to the north of the extinct ridge and 'W' shaped lineations that record back and forth ridge propagation. Microplate formation provides a precise means of dating the onset of the India-Eurasia collision, and is completely independent of and complementary to timing

GMR-based PhC biosensor: FOM analysis and experimental studies

DOE Office of Scientific and Technical Information (OSTI.GOV)

Syamprasad, Jagadeesh; Narayanan, Roshni; Joseph, Joby

2014-02-20

Guided Mode Resonance based Photonic crystal biosensor has a lot of potential applications. In our work, we are trying to improve their figure of merit values in order to achieve an optimum level through design and fabrication techniques. A robust and low-cost alternative for current biosensors is also explored through this research.

Counter-Rotating Magellan and Trinidad Microplates at the Mesozoic Pacific-Phoenix-Farallon Triple Junction

NASA Astrophysics Data System (ADS)

Schouten, H.; Smith, D. K.

2005-12-01

Magellan and Trinidad microplates developed at the Mesozoic triple junction between the Pacific, Phoenix and Farallon plates; the microplates were instrumental in the transition from a transform-ridge-transform to a ridge-ridge-ridge triple junction, which took several tens of millions of years. Contrasting qualitative models for the evolution of these microplates [e.g., Tamaki and Larson, 1988; Nakanishi et al., 1992] provide meager insight in the mechanics of microplate evolution and triple junction transformation. We propose a quantitative model for the evolution of Magellan and Trinidad microplates based on the edge-driven microplate kinematic principles [Schouten et al., 1993] that have provided successful quantitative solutions for the motions of Easter, Juan Fernandez, and Galapagos microplates. In these edge-driven solutions, two angular velocity vectors (describing motion between microplate and driving plates) are located on the microplate boundaries at the tip of rifts that propagate between microplate and driving plates. The rift propagation leaves pseudofaults on microplate and driving plates; the pseudofaults, which can be recognized in the seafloor topography, then become proxies for the trajectories of the angular velocity vectors from which a quantitative solution of microplate motion is derived. Using the estimated seafloor topography of the region and published marine magnetic anomaly lineations we propose the following scenario. The Magellan microplate rotated counterclockwise as evidenced by the fanning of magnetic lineations about the Magellan Trough and the rotation of the older Mid-Pac Mountains lineation set. The Trinidad microplate rotated clockwise relative to the Pacific plate to judge from the wedge-shaped region about the Trinidad trough that has its narrow tip on the Victoria fracture zone (recognized in the estimated seafloor topograpy). The clockwise motion of the Trinidad microplate was driven by Pacific-Phoenix motion; the

A continuous perfusion microplate for cell culture.

PubMed

Goral, Vasiliy N; Zhou, Chunfeng; Lai, Fang; Yuen, Po Ki

2013-03-21

We describe a 96-well microplate with fluidically connected wells that enables the continuous fluid perfusion between wells without the need for external pumping. A single unit in such a perfusion microplate consists of three wells: a source well, a sample (cell culture) well in the middle and a waste well. Fluid perfusion is achieved using a combination of the hydrostatic pressure generated by different liquid levels in the wells and the fluid wicking through narrow strips of a cellulose membrane connecting the wells. There is an excellent correspondence between the observed perfusion flow dynamics and the flow simulations based on Darcy's Law. Hepatocytes (C3A cells) cultured for 4 days in the perfusion microplate with no media exchange in the cell culture well had the same viability as hepatocytes exposed to a daily exchange of media. EOC 20 cells that require media conditioned by LADMAC cells were shown to be equally viable in the adjacent cell culture well of the perfusion microplate with LADMAC cells cultured in the source well. Tegafur, a prodrug, when added to primary human hepatocytes in the source well, was metabolized into a cytotoxic metabolite that kills colon cancer cells (HCT 116) cultured in the adjacent cell culture well; no toxicity was observed when only medium was in the source well. These results suggest that the perfusion microplate is a useful tool for a variety of cell culture applications with benefits ranging from labor savings to enabling in vivo-like toxicity studies.

Structural patterns and tectonic history of the Bauer microplate, Eastern Tropical Pacific

USGS Publications Warehouse

Eakins, B.W.; Lonsdale, P.F.

2003-01-01

The Bauer microplate was an independent slab of oceanic lithosphere that from 17 Ma to 6 Ma grew from 1.4 ?? 105 km2 to 1.2 ?? 106 km2 between the rapidly diverging Pacific and Nazca plates. Growth was by accretion at the lengthening and overlapping axes of the (Bauer-Nazca) Galapagos Rise (GR) and the (Pacific-Bauer) East Pacific Rise (EPR). EPR and GR axial propagation to create and rapidly grow the counter-clockwise spinning microplate occurred in two phases: (1) 17-15Ma, when the EPR axis propagated north and the GR axis propagated south around a narrow (100- to 200-km-wide) core of older lithosphere; and (2) 8-6 Ma, when rapid northward propagation of the EPR axis resumed, overlapping ???400 km of the fast-spreading Pacific-Nazca rise-crest and appending a large (200- to 400-km-wide) area of the west flank of that rise as a 'northern annex' to the microplate. Between 15 and 8 Ma the microplate grew principally by crustal accretion at the crest of its rises. The microplate was captured by the Nazca plate and the Galapagos Rise axis became extinct soon after 6 Ma, when the south end of the Pacific-Bauer EPR axis became aligned with the southern Pacific-Nazca EPR axis and its north end was linked by the Quebrada Transform to the northern Pacific-Nazca EPR axis. Incomplete multibeam bathymetry of the microplate margins, and of both flanks of the Pacific-Bauer and Bauer-Nazca Rises, together with archival magnetic and satellite altimetry data, clarifies the growth and (counter-clockwise) rotation of the microplate, and tests tectonic models derived from studies of the still active, much smaller, Easter and Juan Fernandez microplates. Our interpretations differ from model predictions in that Euler poles were not located on the microplate boundary, propagation in the 15-8 Ma phase of growth was not toward these poles, and microplate rotation rates were small (5??/m.y.) for much of its history, when long, bounding transform faults reduced coupling to Nazca plate

Label-free in vitro prostate cancer cell detection via photonic-crystal biosensor

NASA Astrophysics Data System (ADS)

DeLuna, Frank; Ding, XiaoFei; Sagredo, Ismael; Bustamante, Gilbert; Sun, Lu-Zhe; Ye, Jing Yong

2018-02-01

Prostate-specific antigen (PSA) biomarker assays are the current clinical method for mass screening of prostate cancer. However, high false-positive rates are often reported due to PSA's low specificity, leading to an urgent need for the development of a more specific detection system independent of PSA levels. In our previous research, we demonstrated the feasibility of using cellular refractive indices (RI) as a unique contrast parameter to accomplish label-free detection of prostate cancer cells via variance testing, but were unable to determine if a specific cell was cancerous or noncancerous. In this paper, we report the use of our Photonic-Crystal biosensor in a Total-Internal-Reflection (PC-TIR) configuration to construct a label-free imaging system, which allows for the detection of individual prostate cancer cells utilizing cellular RI as the only contrast parameter. Noncancerous prostate (BPH-1) cells and prostate cancer (PC-3) cells were mixed at varied ratios and measured concurrently. Additionally, we isolated and induced PC-3 cells to undergo epithelial-mesenchymal transition (EMT) by exposing these cells to soluble factors such as TGF-β1. The biophysical characteristics of the cellular RI were quantified extensively in comparison to non-induced PC-3 cells as well as BPH-1 cells. EMT is a crucial mechanism for the invasion and metastasis of epithelial tumors characterized by the loss of cell-cell adhesion and increased cell mobility. Our study shows promising clinical potential in utilizing the PC-TIR biosensor imaging system to not only detect prostate cancer cells, but also evaluate prostate cancer progression.

Nano-machining of biosensor electrodes through gold nanoparticles deposition produced by femtosecond laser ablation

NASA Astrophysics Data System (ADS)

Della Ventura, B.; Funari, R.; Anoop, K. K.; Amoruso, S.; Ausanio, G.; Gesuele, F.; Velotta, R.; Altucci, C.

2015-06-01

We report an application of femtosecond laser ablation to improve the sensitivity of biosensors based on a quartz crystal microbalance device. The nanoparticles produced by irradiating a gold target with 527-nm, 300-fs laser pulses, in high vacuum, are directly deposited on the quartz crystal microbalance electrode. Different gold electrodes are fabricated by varying the deposition time, thus addressing how the nanoparticles surface coverage influences the sensor response. The modified biosensor is tested by weighting immobilized IgG antibody from goat and its analyte (IgG from mouse), and the results are compared with a standard electrode. A substantial increase of biosensor sensitivity is achieved, thus demonstrating that femtosecond laser ablation and deposition is a viable physical method to improve the biosensor sensitivity by means of nanostructured electrodes.

Philippine microplate tectonics and hydrocarbon exploration

DOE Office of Scientific and Technical Information (OSTI.GOV)

Gallagher, J.J. Jr.

1986-07-01

Hydrocarbon traps in the Philippine Islands developed during a long, complex history of microplate tectonics. Carbonate and clastic stratigraphic traps formed during Mesozoic and early Cenozoic rifting and drifting. Hydrocarbons, generated in deep rift basins, migrated to the traps during drifting. Later Cenozoic compressional tectonic activity and concomitant faulting enhanced some traps and destroyed others. Seismic data offshore from Palawan Island reveal the early trap histories. Later trap histories can be interpreted from seismic, outcrop, and remote-sensing data. Understanding the microplate tectonic history of the Philippines is the key to interpreting trap histories.

Highly selective detection of single-nucleotide polymorphisms using a quartz crystal microbalance biosensor based on the toehold-mediated strand displacement reaction.

PubMed

Wang, Dingzhong; Tang, Wei; Wu, Xiaojie; Wang, Xinyi; Chen, Gengjia; Chen, Qiang; Li, Na; Liu, Feng

2012-08-21

Toehold-mediated strand displacement reaction (SDR) is first introduced to develop a simple quartz crystal microbalance (QCM) biosensor without an enzyme or label at normal temperature for highly selective and sensitive detection of single-nucleotide polymorphism (SNP) in the p53 tumor suppressor gene. A hairpin capture probe with an external toehold is designed and immobilized on the gold electrode surface of QCM. A successive SDR is initiated by the target sequence hybridization with the toehold domain and ends with the unfolding of the capture probe. Finally, the open-loop capture probe hybridizes with the streptavidin-coupled reporter probe as an efficient mass amplifier to enhance the QCM signal. The proposed biosensor displays remarkable specificity to target the p53 gene fragment against single-base mutant sequences (e.g., the largest discrimination factor is 63 to C-C mismatch) and high sensitivity with the detection limit of 0.3 nM at 20 °C. As the crucial component of the fabricated biosensor for providing the high discrimination capability, the design rationale of the capture probe is further verified by fluorescence sensing and atomic force microscopy imaging. Additionally, a recovery of 84.1% is obtained when detecting the target sequence in spiked HeLa cells lysate, demonstrating the feasibility of employing this biosensor in detecting SNPs in biological samples.

Photosensitizer adhered to cell culture microplates induces phototoxicity in carcinoma cells.

PubMed

Ziegler, Verena; Kiesslich, Tobias; Krammer, Barbara; Plaetzer, Kristjan

2013-01-01

In vitro experiments in plastic receptacles are the basis of characterization of new photosensitizers (PSs) for the photodynamic therapy. We recently reported that lipophilic PSs adhere to cell culture microplates in a kinetic-like manner (Engelhardt et al., 2011). In the current study, we examined the interaction and phototoxic effects of the microplate-adhered PS in cancer cells. Therefore, we preloaded microplates with hypericin, Foscan, PVP-hypericin, or aluminum (III) phthalocyanine tetrasulfonate chloride (AlPCS4) for 24 hours and measured the PS distribution after addition of A431 human carcinoma cells: following another 24 hours up to 68% of hypericin were detected in the cell fraction. The hydrophilic PVP-hypericin and AlPCS4 also diffused into the cells, but the quantities of PS adherence were considerably lower. Microplate-adhered Foscan appeared not to be redistributed. In contrast to the hydrophilic PSs, the cellular phototoxicity of microplate-adhered lipophilic PS was high, independent of whether the PS (i) was pre-loaded onto microplates or (ii) added simultaneously with the cells or (iii) one day after cell seeding. Based on these results, we suggest testing lipophilic PS dyes for their adherence to microplates. Furthermore, the ability of plastic materials to (reversibly) store PSs might represent a new approach for the PS delivery or the development of antimicrobial coatings.

Photosensitizer Adhered to Cell Culture Microplates Induces Phototoxicity in Carcinoma Cells

PubMed Central

Ziegler, Verena; Kiesslich, Tobias; Krammer, Barbara; Plaetzer, Kristjan

2013-01-01

In vitro experiments in plastic receptacles are the basis of characterization of new photosensitizers (PSs) for the photodynamic therapy. We recently reported that lipophilic PSs adhere to cell culture microplates in a kinetic-like manner (Engelhardt et al., 2011). In the current study, we examined the interaction and phototoxic effects of the microplate-adhered PS in cancer cells. Therefore, we preloaded microplates with hypericin, Foscan, PVP-hypericin, or aluminum (III) phthalocyanine tetrasulfonate chloride (AlPCS4) for 24 hours and measured the PS distribution after addition of A431 human carcinoma cells: following another 24 hours up to 68% of hypericin were detected in the cell fraction. The hydrophilic PVP-hypericin and AlPCS4 also diffused into the cells, but the quantities of PS adherence were considerably lower. Microplate-adhered Foscan appeared not to be redistributed. In contrast to the hydrophilic PSs, the cellular phototoxicity of microplate-adhered lipophilic PS was high, independent of whether the PS (i) was pre-loaded onto microplates or (ii) added simultaneously with the cells or (iii) one day after cell seeding. Based on these results, we suggest testing lipophilic PS dyes for their adherence to microplates. Furthermore, the ability of plastic materials to (reversibly) store PSs might represent a new approach for the PS delivery or the development of antimicrobial coatings. PMID:23509741

Forecastable and Guidable Bubble-Propelled Microplate Motors for Cell Transport.

PubMed

Hu, Narisu; Zhang, Bin; Gai, Meiyu; Zheng, Ce; Frueh, Johannes; He, Qiang

2017-06-01

Cell transport is important to renew body functions and organs with stem cells, or to attack cancer cells with immune cells. The main hindrances of this method are the lack of understanding of cell motion as well as proper transport systems. In this publication, bubble-propelled polyelectrolyte microplates are used for controlled transport and guidance of HeLa cells. Cells survive attachment on the microplates and up to 22 min in 5% hydrogen peroxide solution. They can be guided by a magnetic field whereby increased friction of cells attached to microplates decreases the speed by 90% compared to pristine microplates. The motion direction of the cell-motor system is easier to predict due to the cell being opposite to the bubbles. © 2017 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

A biosensor based on photonic crystal surface waves with an independent registration of the liquid refractive index.

PubMed

Konopsky, Valery N; Alieva, Elena V

2010-01-15

A high-precision optical biosensor technique capable of independently determining the refractive index (RI) of liquids is presented. Photonic crystal surface waves were used to detect surface binding events, while an independent registration of the critical angle was used for accurate determination of the liquid RI. This technique was tested using binding of biotin molecules to a streptavidin monolayer at low and high biotin concentrations. The attained baseline noise is 5x10(-13) m/Hz(1/2) for adlayer thickness changes and 9x10(-8) RIU/Hz(1/2) for RI changes. Copyright 2009 Elsevier B.V. All rights reserved.

High throughput screening using acoustic droplet ejection to combine protein crystals and chemical libraries on crystallization plates at high density

DOE PAGES

Teplitsky, Ella; Joshi, Karan; Ericson, Daniel L.; ...

2015-07-01

We describe a high throughput method for screening up to 1728 distinct chemicals with protein crystals on a single microplate. Acoustic droplet ejection (ADE) was used to co-position 2.5 nL of protein, precipitant, and chemicals on a MiTeGen in situ-1 crystallization plate™ for screening by co-crystallization or soaking. ADE-transferred droplets follow a precise trajectory which allows all components to be transferred through small apertures in the microplate lid. The apertures were large enough for 2.5 nL droplets to pass through them, but small enough so that they did not disrupt the internal environment created by the mother liquor. Using thismore » system, thermolysin and trypsin crystals were efficiently screened for binding to a heavy-metal mini-library. Fluorescence and X-ray diffraction were used to confirm that each chemical in the heavy-metal library was correctly paired with the intended protein crystal. Moreover, a fragment mini-library was screened to observe two known lysozyme We describe a high throughput method for screening up to 1728 distinct chemicals with protein crystals on a single microplate. Acoustic droplet ejection (ADE) was used to co-position 2.5 nL of protein, precipitant, and chemicals on a MiTeGen in situ-1 crystallization plate™ for screening by co-crystallization or soaking. ADE-transferred droplets follow a precise trajectory which allows all components to be transferred through small apertures in the microplate lid. The apertures were large enough for 2.5 nL droplets to pass through them, but small enough so that they did not disrupt the internal environment created by the mother liquor. Using this system, thermolysin and trypsin crystals were efficiently screened for binding to a heavy-metal mini-library. Fluorescence and X-ray diffraction were used to confirm that each chemical in the heavy-metal library was correctly paired with the intended protein crystal. A fragment mini-library was screened to observe two known

Microplate-based method for high-throughput screening of microalgae growth potential.

PubMed

Van Wagenen, Jon; Holdt, Susan Løvstad; De Francisci, Davide; Valverde-Pérez, Borja; Plósz, Benedek Gy; Angelidaki, Irini

2014-10-01

Microalgae cultivation conditions in microplates will differ from large-scale photobioreactors in crucial parameters such as light profile, mixing and gas transfer. Hence volumetric productivity (P(v)) measurements made in microplates cannot be directly scaled up. Here we demonstrate that it is possible to use microplates to measure characteristic exponential growth rates and determine the specific growth rate light intensity dependency (μ-I curve), which is useful as the key input for several models that predict P(v). Nannochloropsis salina and Chlorella sorokiniana specific growth rates were measured by repeated batch culture in microplates supplied with continuous light at different intensities. Exponential growth unlimited by gas transfer or self-shading was observable for a period of several days using fluorescence, which is an order of magnitude more sensitive than optical density. The microplate datasets were comparable to similar datasets obtained in photobioreactors and were used an input for the Huesemann model to accurately predict P(v). Copyright © 2014 Elsevier Ltd. All rights reserved.

Absorbance and fluorometric sensing with capillary wells microplates.

PubMed

Tan, Han Yen; Cheong, Brandon Huey-Ping; Neild, Adrian; Liew, Oi Wah; Ng, Tuck Wah

2010-12-01

Detection and readout from small volume assays in microplates are a challenge. The capillary wells microplate approach [Ng et al., Appl. Phys. Lett. 93, 174105 (2008)] offers strong advantages in small liquid volume management. An adapted design is described and shown here to be able to detect, in a nonimaging manner, fluorescence and absorbance assays minus the error often associated with meniscus forming at the air-liquid interface. The presence of bubbles in liquid samples residing in microplate wells can cause inaccuracies. Pipetting errors, if not adequately managed, can result in misleading data and wrong interpretations of assay results; particularly in the context of high throughput screening. We show that the adapted design is also able to detect for bubbles and pipetting errors during actual assay runs to ensure accuracy in screening.

Graphene as a signal amplifier for preparation of ultrasensitive electrochemical biosensors.

PubMed

Filip, Jaroslav; Kasák, Peter; Tkac, Jan

2015-01-01

Early diagnostics of diseases performed with minimal money and time consumption has become achievable due to recent advances in development of biosensors. These devices use biorecognition elements for selective interaction with an analyte and signal readout is obtained via different types of transducers. Operational characteristics of biosensors have been reported to improve substantially, when a diverse range of nanomaterials was employed. This review presents construction of electrochemical biosensors based on graphene, atomically thin 2D carbon crystals, which is currently intensively studied nanomaterial. The most attractive directions of graphene applications in biosensor preparation are discussed here including novel detection and amplification schemes exploiting graphene's unique electrochemical, physical and chemical properties. The future of graphene-based biosensors is most likely bright, but there is still a lot of work to do to fulfill high expectations.

Crude and purified proteasome activity assays are affected by type of microplate.

PubMed

Cui, Ziyou; Gilda, Jennifer E; Gomes, Aldrin V

2014-02-01

Measurement of proteasome activity is fast becoming a commonly used assay in many laboratories. The most common method to measure proteasome activity involves measuring the release of fluorescent tags from peptide substrates in black microplates. Comparisons of black plates used for measuring fluorescence with different properties show that the microplate properties significantly affect the measured activities of the proteasome. The microplate that gave the highest reading of trypsin-like activity of the purified 20S proteasome gave the lowest reading of chymotrypsin-like activity of the 20S proteasome. Plates with medium binding surfaces from two different companies showed an approximately 2-fold difference in caspase-like activity for purified 20S proteasomes. Even standard curves generated using free 7-amino-4-methylcoumarin (AMC) were affected by the microplate used. As such, significantly different proteasome activities, as measured in nmol AMC released/mg/min, were obtained for purified 20S proteasomes as well as crude heart and liver samples when using different microplates. The naturally occurring molecule betulinic acid activated the chymotrypsin-like proteasome activity in three different plates but did not affect the proteasome activity in the nonbinding surface microplate. These findings suggest that the type of proteasome activity being measured and sample type are important when selecting a microplate. Copyright © 2013 Elsevier Inc. All rights reserved.

Absorbance and fluorometric sensing with capillary wells microplates

DOE Office of Scientific and Technical Information (OSTI.GOV)

Tan, Han Yen; Cheong, Brandon Huey-Ping; Neild, Adrian

2010-12-15

Detection and readout from small volume assays in microplates are a challenge. The capillary wells microplate approach [Ng et al., Appl. Phys. Lett. 93, 174105 (2008)] offers strong advantages in small liquid volume management. An adapted design is described and shown here to be able to detect, in a nonimaging manner, fluorescence and absorbance assays minus the error often associated with meniscus forming at the air-liquid interface. The presence of bubbles in liquid samples residing in microplate wells can cause inaccuracies. Pipetting errors, if not adequately managed, can result in misleading data and wrong interpretations of assay results; particularly inmore » the context of high throughput screening. We show that the adapted design is also able to detect for bubbles and pipetting errors during actual assay runs to ensure accuracy in screening.« less

Immobilization of E. coli with autodisplayed Z-domains to a surface-modified microplate for immunoassay.

PubMed

Yoo, Gu; Park, Min; Lee, Eun-Hang; Jose, Joachim; Pyun, Jae-Chul

2011-11-30

Escherichia coli with autodisplayed Z-domains was reported to improve the sensitivity of immunoassays by the orientation control of antibodies. In this work, a sensitive microplate-based immunoassay is presented by immobilizing E. coli cells to a surface-modified microplate. The microplate was prepared by coating parylene-H film with formyl groups, and then covalently coupling poly-L-lysine to the parylene-H film. The E. coli cells were bound to the microplate by charge interactions between the negatively charged E. coli outer membrane and the positively charged microplate surface. In this work, the preparation of the microplate coated with poly-L-lysine is presented. The immobilization efficiency of E. coli to the modified surface was estimated to be far higher than non-specific interaction by fluorescence microscope and the optical transmittance of the modified microplate was measured to be feasible for immunoassay. The microplate-based immunoassay is demonstrated to be feasible for medical diagnosis of inflammatory diseases by using C-reactive protein as a target analyte for the medical diagnosis of inflammatory diseases. Copyright © 2011 Elsevier B.V. All rights reserved.

Slow light Mach-Zehnder interferometer as label-free biosensor with scalable sensitivity

DOE PAGES

Qin, Kun; Hu, Shuren; Retterer, Scott T.; ...

2016-02-05

Our design, fabrication, and characterization of a label-free Mach–Zehnder interferometer (MZI) optical biosensor that incorporates a highly dispersive one-dimensional (1D) photonic crystal in one arm are presented. The sensitivity of this slow light MZI-based sensor scales with the length of the slow light photonic crystal region. The numerically simulated sensitivity of a MZI sensor with a 16 μm long slow light region is 115,000 rad/RIU-cm, which is sevenfold higher than traditional MZI biosensors with millimeter-length sensing regions. Moreover, the experimental bulk refractive index detection sensitivity of 84,000 rad/RIU-cm is realized and nucleic acid detection is also demonstrated.

Enhancement of Fluorescence-Based Sandwich Immunoassay Using Multilayered Microplates Modified with Plasma-Polymerized Films

PubMed Central

Yano, Kazuyoshi; Iwasaki, Akira

2016-01-01

A functional modification of the surface of a 96-well microplate coupled with a thin layer deposition technique is demonstrated for enhanced fluorescence-based sandwich immunoassays. The plasma polymerization technique enabling the deposition of organic thin films was employed for the modification of the well surface of a microplate. A silver layer and a plasma-polymerized film were consecutively deposited on the microplate as a metal mirror and the optical interference layer, respectively. When Cy3-labeled antibody was applied to the wells of the resulting multilayered microplate without any immobilization step, greatly enhanced fluorescence was observed compared with that obtained with the unmodified one. The same effect could be also exhibited for an immunoassay targeting antigen directly adsorbed on the multilayered microplate. Furthermore, a sandwich immunoassay for the detection of interleukin 2 (IL-2) was performed with the multilayered microplates, resulting in specific and 88-fold–enhanced fluorescence detection. PMID:28029144

Magmatic evolution of the Easter microplate-Crough Seamount region (South East Pacific)

USGS Publications Warehouse

Hekinian, R.; Stoffers, P.; Akermand, D.; Binard, N.; Francheteau, Jean; Devey, C.; Garbe-Schonberg, D.

1995-01-01

The Easter microplate-Crough Seamount region located between 25?? S-116?? W and 25?? S-122?? W consists of a chain of seamounts forming isolated volcanoes and elongated (100-200 km in length) en echelon volcanic ridges oriented obliquely NE (N 065??), to the present day general spreading direction (N 100??) of the Pacific-Nazca plates. The extension of this seamount chain into the southwestern edge of the Easter microplate near 26??30??? S-115?? W was surveyed and sampled. The southern boundary including the Orongo fracture zone and other shallow ridges ( 0.25) MORBs which are similar in composition to other more recent basalts from the Southwest and East Rifts spreading axes of the Easter microplate. Incompatible element ratios normalized to chondrite values [(Ce/Yb)N = 1-2.5}, {(La/Sm)N = 0.4-1.2} and {(Zr/Y)N = 0.7-2.5} of the basalts are also similar to present day volcanism found in the Easter microplate. The volcanics from the Easter microplate-Crough region are unrelated to other known South Pacific intraplate magmatism (i.e. Society, Pitcairn, and Salas y Gomez Islands). Instead their range in incompatible element ratios is comparable to the submarine basalts from the recently investigated Ahu and Umu volcanic field (Easter hotspot) (Scientific Party SO80, 1993) and centered at about 80 km west of Easter Island. The oblique ridges and their associated seamounts are likely to represent ancient leaky transform faults created during the initial stage of the Easter microplate formation (??? 5 Ma). It appears that volcanic activity on seamounts overlying the oblique volcanic ridges has continued during their westward drift from the microplate as shown by the presence of relatively fresh lava observed on one of these structures, namely the first Oblique Volcanic Ridge near 25?? S-118?? W at about 160 km west of the Easter microplate West Rift. Based on a reconstruction of the Easter microplate, it is suggested that the Crough seamount (< 800 m depth) was formed

Crustal Accretion and Mantle Geodynamics at Microplates: Constraints from Gravity Analysis

NASA Astrophysics Data System (ADS)

Ames, K.; Georgen, J. E.; Dordevic, M. M.

2013-12-01

Oceanic crustal accretion occurs in a variety of locations, including mid-ocean ridges and back-arc spreading centers, and in unique settings within these systems, such as plate boundary triple junctions, intra-transform spreading centers, and microplates. This study focuses on crustal accretion and mantle geodynamics at microplates. The Easter and Juan Fernandez microplates are located in the South Pacific along the Pacific, Nazca and Antarctic plate boundaries. Both microplates formed 3-5 Ma and they are currently rotating clockwise at 15 deg/Ma and 9 deg/Ma respectively (e.g., Searle et al. J. Geol. Soc. Lond. 1993). The study area also encompasses the Easter/Sala y Gomez mantle plume and the Foundation seamount chain, both of which are located close to spreading centers. We calculate mantle Bouguer anomaly (MBA) from satellite gravity measurements and shipboard soundings in order to gain a better understanding of the thermal structure of these two oceanic microplates and to quantify the effect that melting anomalies may have on their boundaries. We assume a crustal thickness of 6.0 km, a 1.7 g/cm^3 density difference at the water/crust interface, and a 0.6 g/cm^3 density difference at the crust/mantle interface. The west rift of the Easter microplate has an MBA low ranging from approximately -50 to -100 mGal, while the east rift has slightly higher MBA values ranging from roughly 10 to -50 mGal. The west rift of the Juan Fernandez microplate has a maximum MBA low of about -100 mGal with a sharp increase to -20 mGal at -35 deg S. The east rift of the Juan Fernandez microplate is characterized by more variable MBA, ranging from 0 to -140 mGal. The MBA low associated with the Easter/Sala y Gomez mantle plume has a maximum amplitude about 150 mGal. Likewise, the Foundation seamounts show a gravity low of -140 to -150 mGal. These spatial variations in gravity, as well as published isotopic data and exploratory numerical models, are used to constrain upper mantle

Customizable PCR-microplate array for differential identification of multiple pathogens

PubMed Central

Woubit, Abdela; Yehualaeshet, Teshome; Roberts, Sherrelle; Graham, Martha; Kim, Moonil; Samuel, Temesgen

2014-01-01

Customizable PCR-microplate arrays were developed for the rapid identification of Francisella tularensis subsp. tularensis, Salmonella Typhi, Shigella dysenteriae, Yersinia pestis, Vibrio cholerae Escherichia coli O157:H7, Salmonella Typhimurium, Salmonella Saintpaul, Francisella tularensis subsp. novicida, Vibrio parahaemolyticus, and Yersinia pseudotuberculosis. Previously, we identified highly specific primers targeting each of the pathogens above. Here, we report the development of customizable PCR-microplate arrays for simultaneous identification of the pathogens using the primers. A mixed aliquot of genomic DNA from 38 different strains was used to validate three PCR-microplate array formats. Identical PCR conditions were used to run all the samples on the three formats. Results show specific amplifications on all the three custom plates. In a preliminary test to evaluate the sensitivity of these assays in laboratory-inoculated samples, detection limits as low as 9 cfu/g/ml S. Typhimurium were obtained from beef hot dog, and 78 cfu/ml from milk. Such microplate arrays could serve as valuable tools for initial identification or secondary confirmation of these pathogens. PMID:24215700

Customizable PCR-microplate array for differential identification of multiple pathogens.

PubMed

Woubit, Abdela; Yehualaeshet, Teshome; Roberts, Sherrelle; Graham, Martha; Kim, Moonil; Samuel, Temesgen

2013-11-01

Customizable PCR-microplate arrays were developed for the rapid identification of Salmonella Typhimurium, Salmonella Saintpaul, Salmonella Typhi, Shigella dysenteriae, Escherichia coli O157:H7, Francisella tularensis subsp. tularensis, Francisella tularensis subsp. novicida, Vibrio cholerae, Vibrio parahaemolyticus, Yersinia pestis, and Yersinia pseudotuberculosis. Previously, we identified highly specific primers targeting each of these pathogens. Here, we report the development of customizable PCR-microplate arrays for simultaneous identification of the pathogens using the primers identified. A mixed aliquot of genomic DNA from 38 strains was used to validate three PCR-microplate array formats. Identical PCR conditions were used to run all the samples on the three formats. Specific amplifications were obtained on all three custom plates. In preliminary tests performed to evaluate the sensitivity of these assays in samples inoculated in the laboratory with Salmonella Typhimurium, amplifications were obtained from 1 g of beef hot dog inoculated at as low as 9 CFU/ml or from milk inoculated at as low as 78 CFU/ml. Such microplate arrays could be valuable tools for initial identification or secondary confirmation of contamination by these pathogens.

Development of LEDs-based microplate reader for bioanalytical assay measurements

NASA Astrophysics Data System (ADS)

Alaruri, Sami D.; Katzlinger, Michael; Schinwald, Bernhard; Kronberger, Georg; Atzler, Joseph

2013-10-01

The optical design for an LEDs-based microplate reader that can perform fluorescence intensity (top and bottom), absorbance, luminescence and time-resolved fluorescence measurements is described. The microplate reader is the first microplate reader in the marketplace that incorporates LEDs as excitation light sources. Absorbance measurements over the 0-3.5 optical density range for caffeine solution are presented. Additionally, fluorescence intensity readings collected at 535 and 625 nm from a green and a red RediPlateTM are reported. Furthermore, fluorescence decay lifetime measurements obtained for Eu (europium) and Sm (samarium) standard solutions using 370 nm excitation are presented. The microplate reader detection limits for the fluorescence intensity top, fluorescence intensity bottom, fluorescence polarization and time-resolved fluorescence modes are 1.5 fmol 100 µL-1 fluorescein (384-well plate), 25 fmol 100 µL-1 fluorescein (384-well plate), 5 mP at 10 nM fluorescein (black 384-well plate) and 30 amol 100 µL-1 europium solution (white 384-well plate), respectively.

Saddle-nose deformity repair with microplate-adapted costal cartilage.

PubMed

Eren, Fikret; Öksüz, Sinan; Melikoğlu, Cenk; Karagöz, Hüseyin; Ülkür, Ersin

2014-08-01

Nasal deformities affecting the bone and lower two-thirds of the nose due to the loss of septal height and tip support are defined as "saddle-nose" deformity. Reconstruction of a saddle-nose deformity essentially necessitates structural grafting. This article presents an alternative approach for correction of saddle-nose deformity using a microplate and costal cartilage. The results are compared with those of the previously applied costal cartilage repair methods. Between 2004 and 2013, 16 patients were treated with costal cartilage autografts. Of these 16 patients, 7 were treated with a microplate and costal cartilage autograft combination, 4 were treated with a costal cartilage autograft and Kirschner (K)-wire, and 5 were treated with onlay costal cartilage grafts. The mean follow-up periods were 16 months for group treated with microplate-adapted autologous costal cartilage, 12 months for the group treated with K-wire and autologous costal cartilage, and 16 months for the group treated with onlay costal cartilage. The patients treated with K-wire inserted cartilages and the patients treated onlay dorsal costal cartilages encountered complications such as extrusion of the wire and warping, respectively. The seven patients treated with microplate and dorsal onlay costal cartilage graft did not experience any infection, warping, or extrusion complication. The warping tendency of the costal cartilage autograft can be efficiently prevented without a prominent complication risk by using microplate-adapted costal cartilage grafts. This journal requires that authors assign a level of evidence to each article. For a full description of these Evidence-Based Medicine ratings, please refer to the Table of Contents or the online Instructions to Authors www.springer.com/00266 .

Determination of High-affinity Antibody-antigen Binding Kinetics Using Four Biosensor Platforms.

PubMed

Yang, Danlin; Singh, Ajit; Wu, Helen; Kroe-Barrett, Rachel

2017-04-17

Label-free optical biosensors are powerful tools in drug discovery for the characterization of biomolecular interactions. In this study, we describe the use of four routinely used biosensor platforms in our laboratory to evaluate the binding affinity and kinetics of ten high-affinity monoclonal antibodies (mAbs) against human proprotein convertase subtilisin kexin type 9 (PCSK9). While both Biacore T100 and ProteOn XPR36 are derived from the well-established Surface Plasmon Resonance (SPR) technology, the former has four flow cells connected by serial flow configuration, whereas the latter presents 36 reaction spots in parallel through an improvised 6 x 6 crisscross microfluidic channel configuration. The IBIS MX96 also operates based on the SPR sensor technology, with an additional imaging feature that provides detection in spatial orientation. This detection technique coupled with the Continuous Flow Microspotter (CFM) expands the throughput significantly by enabling multiplex array printing and detection of 96 reaction sports simultaneously. In contrast, the Octet RED384 is based on the BioLayer Interferometry (BLI) optical principle, with fiber-optic probes acting as the biosensor to detect interference pattern changes upon binding interactions at the tip surface. Unlike the SPR-based platforms, the BLI system does not rely on continuous flow fluidics; instead, the sensor tips collect readings while they are immersed in analyte solutions of a 384-well microplate during orbital agitation. Each of these biosensor platforms has its own advantages and disadvantages. To provide a direct comparison of these instruments' ability to provide quality kinetic data, the described protocols illustrate experiments that use the same assay format and the same high-quality reagents to characterize antibody-antigen kinetics that fit the simple 1:1 molecular interaction model.

Improvement of up-converting phosphor technology-based biosensor

NASA Astrophysics Data System (ADS)

Xie, Chengke; Huang, Lihua; Zhang, Youbao; Guo, Xiaoxian; Qu, Jianfeng; Huang, Huijie

2008-12-01

A novel biosensor based on up-converting phosphor technology (UPT) was developed several years ago. It is a kind of optical biosensor using up-converting phosphor (UCP) particles as the biological marker. From then on, some improvements have been made for this UPT-based biosensor. The primary aspects of the improvement lie in the control system. On one hand, the hardware of the control system has been optimized, including replacing two single chip microcomputers (SCM) with only one, the optimal design of the keyboard interface circuit and the liquid crystal module (LCM) control circuit et al.. These result in lower power consumption and higher reliability. On the other hand, a novel signal processing algorithm is proposed in this paper, which can improve the automation and operating simplicity of the UPT-based biosensor. It has proved to have high sensitivity (~ng/ml), high stability and good repeatability (CV<5%), which is better than the former system. It can meet the need of some various applications such as rapid immunoassay, chemical and biological detection and so on.

Novel versatile smart phone based Microplate readers for on-site diagnoses.

PubMed

Fu, Qiangqiang; Wu, Ze; Li, Xiuqing; Yao, Cuize; Yu, Shiting; Xiao, Wei; Tang, Yong

2016-07-15

Microplate readers are important diagnostic instruments, used intensively for various readout test kits (biochemical analysis kits and ELISA kits). However, due to their expensive and non-portability, commercial microplate readers are unavailable for home testing, community and rural hospitals, especially in developing countries. In this study, to provide a field-portable, cost-effective and versatile diagnostic tool, we reported a novel smart phone based microplate reader. The basic principle of this devise relies on a smart phone's optical sensor that measures transmitted light intensities of liquid samples. To prove the validity of these devises, developed smart phone based microplate readers were applied to readout results of various analytical targets. These targets included analanine aminotransferase (ALT; limit of detection (LOD) was 17.54 U/L), alkaline phosphatase (AKP; LOD was 15.56 U/L), creatinine (LOD was 1.35μM), bovine serum albumin (BSA; LOD was 0.0041mg/mL), prostate specific antigen (PSA; LOD was 0.76pg/mL), and ractopamine (Rac; LOD was 0.31ng/mL). The developed smart phone based microplate readers are versatile, portable, and inexpensive; they are unique because of their ability to perform under circumstances where resources and expertize are limited. Copyright © 2016 Elsevier B.V. All rights reserved.

Measurement of filter paper activities of cellulase with microplate-based assay.

PubMed

Yu, Xiaoxiao; Liu, Yan; Cui, Yuxiao; Cheng, Qiyue; Zhang, Zaixiao; Lu, Jia Hui; Meng, Qingfan; Teng, Lirong; Ren, Xiaodong

2016-01-01

It is always a challenge to determine the total cellulase activity efficiently without reducing accuracy. The most common total cellulase activity assay is the filter paper assay (FPA) established by the International Union of Pure and Applied Chemistry (IUPAC). A new procedure to measure the FPA with microplate-based assay was studied in this work, which followed the main idea of IUPAC to dilute cellulase preparation to get fixed glucose release. FPAs of six cellulase preparations were determined with the microplate-based assay. It is shown that FPAs of cellulase Youtell, RCconc, R-10, Lerkam, Yishui and Sinopharm were 67.9, 46.0, 46.1, 27.4, 7.6 and 8.0 IU/ml respectively. There was no significant difference at the 95% confidence level between the FPA determined with IUPAC and the microplate-based assay. It could be concluded that the FPA could be determined by the microplate-based assay with the same accuracy and much more efficiency compared with that by IUPAC.

Measurement of filter paper activities of cellulase with microplate-based assay

PubMed Central

Yu, Xiaoxiao; Liu, Yan; Cui, Yuxiao; Cheng, Qiyue; Zhang, Zaixiao; Lu, Jia Hui; Meng, Qingfan; Teng, Lirong; Ren, Xiaodong

2015-01-01

It is always a challenge to determine the total cellulase activity efficiently without reducing accuracy. The most common total cellulase activity assay is the filter paper assay (FPA) established by the International Union of Pure and Applied Chemistry (IUPAC). A new procedure to measure the FPA with microplate-based assay was studied in this work, which followed the main idea of IUPAC to dilute cellulase preparation to get fixed glucose release. FPAs of six cellulase preparations were determined with the microplate-based assay. It is shown that FPAs of cellulase Youtell, RCconc, R-10, Lerkam, Yishui and Sinopharm were 67.9, 46.0, 46.1, 27.4, 7.6 and 8.0 IU/ml respectively. There was no significant difference at the 95% confidence level between the FPA determined with IUPAC and the microplate-based assay. It could be concluded that the FPA could be determined by the microplate-based assay with the same accuracy and much more efficiency compared with that by IUPAC. PMID:26858572

Nanophotonic label-free biosensors for environmental monitoring.

PubMed

Chocarro-Ruiz, Blanca; Fernández-Gavela, Adrián; Herranz, Sonia; Lechuga, Laura M

2017-06-01

The field of environmental monitoring has experienced a substantial progress in the last years but still the on-site control of contaminants is an elusive problem. In addition, the growing number of pollutant sources is accompanied by an increasing need of having efficient early warning systems. Several years ago biosensor devices emerged as promising environmental monitoring tools, but their level of miniaturization and their fully operation outside the laboratory prevented their use on-site. In the last period, nanophotonic biosensors based on evanescent sensing have emerged as an outstanding choice for portable point-of-care diagnosis thanks to their capability, among others, of miniaturization, multiplexing, label-free detection and integration in lab-on-chip platforms. This review covers the most relevant nanophotonic biosensors which have been proposed (including interferometric waveguides, grating-couplers, microcavity resonators, photonic crystals and localized surface plasmon resonance sensors) and their recent application for environmental surveillance. Copyright © 2017 Elsevier Ltd. All rights reserved.

Glucose biosensor based on GOx/HRP bienzyme at liquid-crystal/aqueous interface.

PubMed

Khan, Mashooq; Park, Soo-Young

2015-11-01

Glucose oxidase (GOx) and horseradish peroxidase (HRP) were co-immobilized to the polyacrylicacid block of a poly(acrylicacid-b-4-cyanobiphenyl-4'-undecylacrylate) (PAA-b-LCP) copolymer in water. PAA-b-LCP was strongly anchored by the LCP block in 4-cyano-4'-pentylbiphenyl (5CB) which was contained in a transmission electron microscope (TEM) grid for glucose detection. The optimal conditions for the performance of the TEM grid glucose biosensor were studied in terms of the activity and stability of the immobilized enzymes. Glucose in water was detected by the 5CB changing from a planar to a homeotropic orientation, as observed through a polarized optical microscope. The TEM biosensor detected glucose concentrations at ⩾0.02 mM, with an optimal GOx/HRP molar ratio of 3/1. This glucose biosensor has characteristics of enzyme sensitivity and stability, reusability, the ease and selective glucose detection which may provide a new way of detecting glucose. Copyright © 2015 Elsevier Inc. All rights reserved.

Euler-Vector Clustering of GPS Velocities Defines Microplate Geometry in Southwest Japan

NASA Astrophysics Data System (ADS)

Savage, J. C.

2018-02-01

I have used Euler-vector clustering to assign 469 GEONET stations in southwest Japan to k clusters (k = 2, 3,..., 9) so that, for any k, the velocities of stations within each cluster are most consistent with rigid-block motion on a sphere. That is, I attempt to explain the raw (i.e., uncorrected for strain accumulation), 1996-2006 velocities of those 469 Global Positioning System stations by rigid motion of k clusters on the surface of a spherical Earth. Because block geometry is maintained as strain accumulates, Euler-vector clustering may better approximate the block geometry than the values of the associated Euler vectors. The microplate solution for each k is constructed by merging contiguous clusters that have closely similar Euler vectors. The best solution consists of three microplates arranged along the Nankaido Trough-Ryukyu Trench between the Amurian and Philippine Sea Plates. One of these microplates, the South Kyushu Microplate (an extension of the Ryukyu forearc into the southeast corner of Kyushu), had previously been identified from paleomagnetic rotations. Relative to ITRF2000 the three microplates rotate at different rates about neighboring poles located close to the northwest corner of Shikoku. The microplate model is identical to that proposed in the block model of Wallace et al. (2009, https://doi.org/10.1130/G2522A.1) except in southernmost Kyushu. On Shikoku and Honshu, but not Kyushu, the microplate model is consistent with that proposed in the block models of Nishimura and Hashimoto (2006, https://doi.org/10.1016/j.tecto.2006.04.017) and Loveless and Meade (2010, https://doi.org/10.1029/2008JB006248) without the low-slip-rate boundaries proposed in the latter.

Optical biosensors.

PubMed

Damborský, Pavel; Švitel, Juraj; Katrlík, Jaroslav

2016-06-30

Optical biosensors represent the most common type of biosensor. Here we provide a brief classification, a description of underlying principles of operation and their bioanalytical applications. The main focus is placed on the most widely used optical biosensors which are surface plasmon resonance (SPR)-based biosensors including SPR imaging and localized SPR. In addition, other optical biosensor systems are described, such as evanescent wave fluorescence and bioluminescent optical fibre biosensors, as well as interferometric, ellipsometric and reflectometric interference spectroscopy and surface-enhanced Raman scattering biosensors. The optical biosensors discussed here allow the sensitive and selective detection of a wide range of analytes including viruses, toxins, drugs, antibodies, tumour biomarkers and tumour cells. © 2016 The Author(s). Published by Portland Press Limited on behalf of the Biochemical Society.

Detection of influenza A virus subtypes using a solid-phase PCR microplate chip assay.

PubMed

Sun, Xin-Cheng; Wang, YunLong; Yang, Liping; Zhang, HuiRu

2015-01-01

A rapid and sensitive microplate chip based on solid PCR was developed to identify influenza A subtypes. A simple ultraviolet cross-linking method was used to immobilize DNA probes on pretreated microplates. Solid-phase PCR was proven to be a convenient method for influenza A screening. The sensitivity of the microplate chip was 10(-3) μg/mL for the enzymatic colorimetric method and 10(-4) μg/mL for the fluorescence method. The 10 sets of primers and probes for the microplate chip were highly specific and did not interfere with each other. These results suggest that the microplate chip based on solid PCR can be used to rapidly detect universal influenza A and its subtypes. This platform can also be used to detect other pathogenic microorganisms. Copyright © 2014 Elsevier B.V. All rights reserved.

Development of Singlet Oxygen Absorption Capacity (SOAC) Assay Method Using a Microplate Reader.

PubMed

Takahashi, Shingo; Iwasaki-Kino, Yuko; Aizawa, Koichi; Terao, Junji; Mukai, Kazuo

2016-01-01

Recently, a new assay method that can quantify the singlet oxygen absorption capacity (SOAC) of natural antioxidants and food extracts was developed. The SOAC values were measured in ethanol-chloroform-D2O (50 + 50 + 1, v/v/v) solution at 35°C using a UV-Vis spectrophotometer equipped with a six-channel cell positioner and an electron-temperature control unit. In the present study, measurement of the SOAC values was performed for eight representative carotenoids and three vegetable extracts (tomato, carrot, and red paprika) using a versatile instrument, the microplate reader. A 24-well glass microplate was used for measurements because a plastic microplate, commonly used in the laboratory, dissolves in the ethanol-chloroform-D2O solution. The SOAC values of eight carotenoids and three vegetable extracts measured using a microplate reader were in good agreement with the corresponding values measured using a UV-Vis spectrophotometer, suggesting that the microplate reader is an applicable instrument for the measurement of reliable SOAC values for general antioxidants and food extracts in solution.

Silaffin peptides as a novel signal enhancer for gravimetric biosensors.

PubMed

Nam, Dong Hyun; Lee, Jeong-O; Sang, Byoung-In; Won, Keehoon; Kim, Yong Hwan

2013-05-01

Application of biomimetic silica formation to gravimetric biosensors has been conducted for the first time. As a model system, silaffin peptides fused with green fluorescent protein (GFP) were immobilized on a gold quartz crystal resonator for quartz crystal microbalances using a self-assembled monolayer. When a solution of silicic acid was supplied, silica particles were successfully deposited on the Au surface, resulting in a significant change in resonance frequency (i.e., signal enhancement) with the silaffin-GFP. However, frequency was not altered when bare GFP was used as a control. The novel peptide enhancer is advantageous because it can be readily and quantitatively conjugated with sensing proteins using recombinant DNA technology. As a proof of concept, this study shows that the silaffin domains can be employed as a novel and efficient biomolecular signal enhancer for gravimetric biosensors.

A novel microfluidic microplate as the next generation assay platform for enzyme linked immunoassays (ELISA).

PubMed

Kai, Junhai; Puntambekar, Aniruddha; Santiago, Nelson; Lee, Se Hwan; Sehy, David W; Moore, Victor; Han, Jungyoup; Ahn, Chong H

2012-11-07

In this work we introduce a novel microfluidic enzyme linked immunoassays (ELISA) microplate as the next generation assay platform for unparalleled assay performances. A combination of microfluidic technology with standard SBS-configured 96-well microplate architecture, in the form of microfluidic microplate technology, allows for the improvement of ELISA workflows, conservation of samples and reagents, improved reaction kinetics, and the ability to improve the sensitivity of the assay by multiple analyte loading. This paper presents the design and characterization of the microfluidic microplate, and its application in ELISA.

Recent advances in rapid pathogen detection method based on biosensors.

PubMed

Chen, Ying; Wang, Zhenzhen; Liu, Yingxun; Wang, Xin; Li, Ying; Ma, Ping; Gu, Bing; Li, Hongchun

2018-06-01

As strain variation and drug resistance become more pervasive, the prevention and control of infection have been a serious problem in recent years. The detection of pathogen is one of the most important parts of the process of diagnosis. Having a series of advantages, such as rapid response, high sensitivity, ease of use, and low cost, biosensors have received much attention and been studied deeply. Moreover, relying on its characteristics of small size, real time, and multiple analyses, biosensors have developed rapidly and used widely and are expected to be applied for microbiological detection in order to meet higher accuracy required by clinical diagnosis. The main goal of this contribution is not to simply collect and list all papers related to pathogen detection based on biosensors published recently, but to discuss critically the development and application of many kinds of biosensors such as electrochemical (amperometric, impedimetric, potentiometric, and conductometric), optical (fluorescent, fibre optic and surface plasmon resonance), and piezoelectric (quartz crystal microbalances and atomic force microscopy) biosensors in pathogen detection as well as the comparisons with the existing clinical detection methods (traditional culture, enzyme-linked immunosorbent assay, polymerase chain reaction, and mass spectrometry).

Wireless poly(dimethylsiloxane) quartz-crystal-microbalance biosensor chip fabricated by nanoimprint lithography for micropump integration aiming at application in lab-on-a-chip

NASA Astrophysics Data System (ADS)

Kato, Fumihito; Noguchi, Hiroyuki; Kodaka, Yukinari; Oshida, Naoya; Ogi, Hirotsugu

2018-07-01

We developed a quartz-crystal-microbalance (QCM) biosensor chip that operates wirelessly via electromagnetic waves, using poly(dimethylsiloxane) (PDMS). An AT-cut quartz oscillator (22–30 µm) is packaged in a microchannel, where it is supported by micropillars without mechanical fixing. As a result, the quartz oscillator is little affected by the thermal stress caused by the difference in the thermal expansion coefficients of the components, and the leakage of the vibration energy of the quartz oscillator is reduced. Consequently, high-frequency (∼56 MHz) measurement with a stable baseline (±∼2 ppm) is realized. We succeeded in repeatedly monitoring the binding reaction between immunoglobulin G (IgG) and Staphylococcus aureus protein A (SPA) with the quartz oscillator on which SPA molecules were immobilized nonspecifically. In addition, the affinity between SPA and IgG was calculated from the association and dissociation curves, and the usefulness of our wireless PDMS QCM biosensor was demonstrated.

A Novel Cell-Based Hybrid Acoustic Wave Biosensor with Impedimetric Sensing Capabilities

PubMed Central

Liu, Fei; Li, Fang; Nordin, Anis Nurashikin; Voiculescu, Ioana

2013-01-01

A novel multiparametric biosensor system based on living cells will be presented. The biosensor system includes two biosensing techniques on a single device: resonant frequency measurements and electric cell-substrate impedance sensing (ECIS). The multiparametric sensor system is based on the innovative use of the upper electrode of a quartz crystal microbalance (QCM) resonator as working electrode for the ECIS technique. The QCM acoustic wave sensor consists of a thin AT-cut quartz substrate with two gold electrodes on opposite sides. For integration of the QCM with the ECIS technique a semicircular counter electrode was fabricated near the upper electrode on the same side of the quartz crystal. Bovine aortic endothelial live cells (BAECs) were successfully cultured on this hybrid biosensor. Finite element modeling of the bulk acoustic wave resonator using COMSOL simulations was performed. Simultaneous gravimetric and impedimetric measurements performed over a period of time on the same cell culture were conducted to validate the device's sensitivity. The time necessary for the BAEC cells to attach and form a compact monolayer on the biosensor was 35∼45 minutes for 1.5 × 104 cells/cm2 BAECs; 60 minutes for 2.0 × 104 cells/cm2 BAECs; 70 minutes for 3.0 × 104 cells/cm2 BAECs; and 100 minutes for 5.0 × 104 cells/cm2 BAECs. It was demonstrated that this time is the same for both gravimetric and impedimetric measurements. This hybrid biosensor will be employed in the future for water toxicity detection. PMID:23459387

[Comparative study on microplate and anchor fixation in open-door cervical expansive laminoplasty].

PubMed

Zeng, Yun; Xiong, Min; Yu, Hualong; He, Ning; Wang, Zhiyong; Liu, Zhigang; Han, Heng; Chen, Sen

2011-08-01

To evaluate the effectiveness of microplate fixation in open-door cervical expansive laminoplasty (ELP) by comparing with anchor fixation. Between January 2005 and October 2008, 35 patients with multi-segment cervical spondylotic myelopathy were treated. Of them, 15 patients underwent ELP by microplate fixation (microplate group) and 20 patients underwent ELP by anchor fixation (anchor group). In microplate group, there were 10 males and 5 females with the age of (51.2 +/- 11.5) years; the disease duration ranged from 6 to 60 months (mean, 14 months); and the preoperative Japanese Orthopaedic Association (JOA) score was 7.7 +/- 2.5. In anchor group, there were 13 males and 7 females with the age of (50.7 +/- 10.8) years; the disease duration ranged from 3 to 58 months (mean, 17 months); and the preoperative JOA score was 7.8 +/- 2.9. There was no significant difference in the general data, such as gender, age, and JOA score between 2 groups (P > 0.05). All incisions healed by first intention. Thirty-five cases were followed up 24-68 months (mean, 32 months). The operation time was (113 +/- 24) minutes in anchor group and (111 +/- 27) minutes in microplate group, showing no significant difference (t = 0.231 3, P = 0.818 5). The rate of spinal canal expansion in microplate group (60% +/- 24%) was significantly higher than that in anchor group (40% +/- 18%) (t = 2.820, P = 0.008). The JOA scores of 2 groups at 3 months and 24 months after operation were significantly higher than the preoperative scores (P < 0.01). There was no significant difference in JOA score between 2 groups at 3 months after operation (t = 1.620 5, P = 0.114 6), but the JOA score of microplate group was significantly higher than that of anchor group at 24 months after operation (t = 3.454 3, P = 0.001 5). X-ray film, MRI, and CT scan at 3-6 months after operation displayed that door spindle reached bony fusion. There was no occurrence of "re-close of door" in 2 groups. The rate of complication in

Whole-cell biosensor of cellobiose and application to wood decay detection.

PubMed

Toussaint, Maxime; Bontemps, Cyril; Besserer, Arnaud; Hotel, Laurence; Gérardin, Philippe; Leblond, Pierre

2016-12-10

Fungal biodegradation of wood is one of the main threats regarding its use as a material. So far, the detection of this decaying process is empirically assessed by loss of mass, when the fungal attack is advanced and woody structure already damaged. Being able to detect fungal attack on wood in earlier steps is thus of special interest for the wood economy. In this aim, we designed here a new diagnostic tool for wood degradation detection based on the bacterial whole-cell biosensor technology. It was designed in diverting the soil bacteria Streptomyces CebR sensor system devoted to cellobiose detection, a cellulolytic degradation by-product emitted by lignolytic fungi since the onset of wood decaying process. The conserved regulation scheme of the CebR system among Streptomyces allowed constructing a molecular tool easily transferable in different strains or species and enabling the screen for optimal host strains for cellobiose detection. Assays are performed in microplates using one-day culture lysates. Diagnostic is performed within one hour by a spectrophotometric measuring of the cathecol deshydrogenase activity. The selected biosensor was able to detect specifically cellobiose at concentrations similar to those measured in decaying wood and in a spruce leachate attacked by a lignolytic fungus, indicating a high potential of applicability to detect ongoing wood decay process. Copyright © 2016 Elsevier B.V. All rights reserved.

Correction of microplate location effects improves performance of the thrombin generation test

PubMed Central

2013-01-01

Background Microplate-based thrombin generation test (TGT) is widely used as clinical measure of global hemostatic potential and it becomes a useful tool for control of drug potency and quality by drug manufactures. However, the convenience of the microtiter plate technology can be deceiving: microplate assays are prone to location-based variability in different parts of the microtiter plate. Methods In this report, we evaluated the well-to-well consistency of the TGT variant specifically applied to the quantitative detection of the thrombogenic substances in the immune globulin product. We also studied the utility of previously described microplate layout designs in the TGT experiment. Results Location of the sample on the microplate (location effect) contributes to the variability of TGT measurements. Use of manual pipetting techniques and applications of the TGT to the evaluation of procoagulant enzymatic substances are especially sensitive. The effects were not sensitive to temperature or choice of microplate reader. Smallest location effects were observed with automated dispenser-based calibrated thrombogram instrument. Even for an automated instrument, the use of calibration curve resulted in up to 30% bias in thrombogenic potency assignment. Conclusions Use of symmetrical version of the strip-plot layout was demonstrated to help to minimize location artifacts even under the worst-case conditions. Strip-plot layouts are required for quantitative thrombin-generation based bioassays used in the biotechnological field. PMID:23829491

Correction of microplate location effects improves performance of the thrombin generation test.

PubMed

Liang, Yideng; Woodle, Samuel A; Shibeko, Alexey M; Lee, Timothy K; Ovanesov, Mikhail V

2013-07-05

Microplate-based thrombin generation test (TGT) is widely used as clinical measure of global hemostatic potential and it becomes a useful tool for control of drug potency and quality by drug manufactures. However, the convenience of the microtiter plate technology can be deceiving: microplate assays are prone to location-based variability in different parts of the microtiter plate. In this report, we evaluated the well-to-well consistency of the TGT variant specifically applied to the quantitative detection of the thrombogenic substances in the immune globulin product. We also studied the utility of previously described microplate layout designs in the TGT experiment. Location of the sample on the microplate (location effect) contributes to the variability of TGT measurements. Use of manual pipetting techniques and applications of the TGT to the evaluation of procoagulant enzymatic substances are especially sensitive. The effects were not sensitive to temperature or choice of microplate reader. Smallest location effects were observed with automated dispenser-based calibrated thrombogram instrument. Even for an automated instrument, the use of calibration curve resulted in up to 30% bias in thrombogenic potency assignment. Use of symmetrical version of the strip-plot layout was demonstrated to help to minimize location artifacts even under the worst-case conditions. Strip-plot layouts are required for quantitative thrombin-generation based bioassays used in the biotechnological field.

Tectonics and evolution of the Juan Fernandez microplate at the Pacific-Nazca-Antarctic triple junction

NASA Technical Reports Server (NTRS)

Anderson-Fontana, S.; Larson, R. L.; Engein, J. F.; Lundgren, P.; Stein, S.

1986-01-01

Magnetic and bathymetric profiles derived from the R/V Endeavor survey and focal mechanism studies for earthquakes on two of the Juan Fernandez microplate boundaries are analyzed. It is observed that the Nazca-Juan Fernandez pole is in the northern end of the microplate since the magnetic lineation along the East Ridge of the microplate fans to the south. The calculation of the relative motion of the Juan Fernandez-Pacific-Nazca-Antarctic four-plate system using the algorithm of Minster et al. (1974) is described. The development of tectonic and evolutionary models of the region is examined. The tectonic model reveals that the northern boundary of the Juan Fernandez microplate is a zone of compression and that the West Ridge and southwestern boundary are spreading obliquely; the evolutionary model relates the formation of the Juan Fernandez microplate to differential spreading rates at the triple junction.

Finite element analysis to determine the stress distribution, displacement and safety factor on a microplate for the fractured jaw case

NASA Astrophysics Data System (ADS)

Pratama, Juan; Mahardika, Muslim

2018-03-01

Microplate is a connecting plate that can be used for jaw bone fixation. In the last two decades, microplate has been used so many times to help reconstruction of fractured jaw bone which is called mandibular bone or mandible bone. The plate is used to provide stable fixation of the fractured bone tissue during healing and reconstruction process. In this study Finite Element Analysis was used to predict the stress concentration and distribution on a microplate, displacement on the microplate and also to determine the safety factor of the microplate based on maximum allowable stress value, and finally to ascertain whether microplate is safe to use or not. The microplate was produced from punching process using titanium grade 1 (pure titanium) as material with a thickness of 500 µm. The results of the research indicated that the microplate was safe to use according to the maximum stress around the hole, displacement around the hole and also the safety factor of the microplate.

Development of a mass sensitive quartz crystal microbalance (QCM)-based DNA biosensor using a 50 MHz electronic oscillator circuit.

PubMed

García-Martinez, Gonzalo; Bustabad, Enrique Alonso; Perrot, Hubert; Gabrielli, Claude; Bucur, Bogdan; Lazerges, Mathieu; Rose, Daniel; Rodriguez-Pardo, Loreto; Fariña, Jose; Compère, Chantal; Vives, Antonio Arnau

2011-01-01

This work deals with the design of a high sensitivity DNA sequence detector using a 50 MHz quartz crystal microbalance (QCM) electronic oscillator circuit. The oscillator circuitry is based on Miller topology, which is able to work in damping media. Calibration and experimental study of frequency noise are carried out, finding that the designed sensor has a resolution of 7.1 ng/cm(2) in dynamic conditions (with circulation of liquid). Then the oscillator is proved as DNA biosensor. Results show that the system is able to detect the presence of complementary target DNAs in a solution with high selectivity and sensitivity. DNA target concentrations higher of 50 ng/mL can be detected.

Specific Detection and Identification of Herpes B Virus by a PCR-Microplate Hybridization Assay

PubMed Central

Oya, Chika; Ochiai, Yoshitsugu; Taniuchi, Yojiro; Takano, Takashi; Ueda, Fukiko; Yoshikawa, Yasuhiro; Hondo, Ryo

2004-01-01

Herpes B virus DNA was specifically amplified by PCR, targeting the regions that did not cross-react with herpes simplex virus (HSV). The amplified products, which were shown to be highly genetic polymorphisms among herpes B virus isolates, were identified by microplate hybridization with probes generated by PCR. The products immobilized in microplate wells were hybridized with the biotin-labeled probes derived from the SMHV strain of herpes B virus. The amplified products derived from the SMHV and E2490 strains of herpes B virus were identified by microplate hybridization. PCR products amplified from the trigeminal ganglia of seropositive cynomolgus macaques were identified as herpes B virus DNA. The utility of the PCR-microplate hybridization assay for genetic detection and identification of the polymorphic region of herpes B virus was determined. PMID:15131142

Liquid crystal-based biosensor with backscattering interferometry: A quantitative approach.

PubMed

Khan, Mashooq; Park, Soo-Young

2017-01-15

We developed a new technology that uses backscattering interferometry (BSI) to quantitatively measure nematic liquid crystal (NLC)-based biosensors, those usually relied on texture reading for on/off signals. The LC-based BSI comprised an octadecyltrichlorosilane (OTS)-coated square capillary filled with 4-cyano-4'-pentylbiphenyl (5CB, a nematic LC at room temperature). The LC/water interface in the capillary was functionalized by a coating of poly(acrylicacid-b-4-cyanobiphenyl-4'-oxyundecylacrylate) (PAA-b-LCP) and immobilized with the enzymes glucose oxidase (GOx) and horseradish peroxidase (HRP) through covalent linkage to the PAA chains (5CB PAA-GOx:HRP ) for glucose detection. Laser irradiation of the LC near the LC/water interface resulted in backscattered fringes with high contrast. The change in the spatial position of the fringes (because of the change in the orientation of the LC caused by the GOx:HRP enzymatic reaction of glucose) altered the output voltage of the photodetector when its active area was aligned with the edge of one of the fringes. The change in the intensity at the photodetector allowed the detection limit of the instrument to be as low as 0.008mM with a linear range of 0.02-9mM in a short response time (~60s). This LC-based BSI technique allows for quantitative, sensitive, selective, reproducible, easily obtainable, and interference-free detection in a large linear dynamic range and for practical applications with human serum. Copyright © 2016 Elsevier B.V. All rights reserved.

Microplate-compatible total internal reflection fluorescence microscopy for receptor pharmacology

NASA Astrophysics Data System (ADS)

Chen, Minghan; Zaytseva, Natalya V.; Wu, Qi; Li, Min; Fang, Ye

2013-05-01

We report the use of total internal reflection fluorescence (TIRF) microscopy for analyzing receptor pharmacology and the development of a microplate-compatible TIRF imaging system. Using stably expressed green fluorescence protein tagged β2-adrenergic receptor as the reporter, we found that the activation of different receptors results in distinct kinetic signatures of the TIRF intensity of cells. These TIRF signatures closely resemble the characteristics of their respective label-free dynamic mass redistribution signals in the same cells. This suggests that TIRF in microplate can be used for profiling and screening drugs.

Recombinant antibodies and their use in biosensors.

PubMed

Zeng, Xiangqun; Shen, Zhihong; Mernaugh, Ray

2012-04-01

Inexpensive, noninvasive immunoassays can be used to quickly detect disease in humans. Immunoassay sensitivity and specificity are decidedly dependent upon high-affinity, antigen-specific antibodies. Antibodies are produced biologically. As such, antibody quality and suitability for use in immunoassays cannot be readily determined or controlled by human intervention. However, the process through which high-quality antibodies can be obtained has been shortened and streamlined by use of genetic engineering and recombinant antibody techniques. Antibodies that traditionally take several months or more to produce when animals are used can now be developed in a few weeks as recombinant antibodies produced in bacteria, yeast, or other cell types. Typically most immunoassays use two or more antibodies or antibody fragments to detect antigens that are indicators of disease. However, a label-free biosensor, for example, a quartz-crystal microbalance (QCM) needs one antibody only. As such, the cost and time needed to design and develop an immunoassay can be substantially reduced if recombinant antibodies and biosensors are used rather than traditional antibody and assay (e.g. enzyme-linked immunosorbant assay, ELISA) methods. Unlike traditional antibodies, recombinant antibodies can be genetically engineered to self-assemble on biosensor surfaces, at high density, and correctly oriented to enhance antigen-binding activity and to increase assay sensitivity, specificity, and stability. Additionally, biosensor surface chemistry and physical and electronic properties can be modified to further increase immunoassay performance above and beyond that obtained by use of traditional methods. This review describes some of the techniques investigators have used to develop highly specific and sensitive, recombinant antibody-based biosensors for detection of antigens in simple or complex biological samples.

Developing Highly Sensitive Micro-Biosensors for in-situ Monitoring Mercury and Chromium(IV) Contaminants by Genetically-evolving and Computer-designing Metal-binding Proteins

DOE Office of Scientific and Technical Information (OSTI.GOV)

Wang, Qinghong; Fang, Xiangdong; Goddard, William

2013-10-17

Mercury has been well known as an environmental pollutant to the environment and to cause serious effects on human health for several decades. To effectively control mercury pollution and reduce mercury damages, the sensitive determination of mercury is essential. Currently, many different types of sensor-based assays have been developed, while the whole-cell biosensor has been gaining increasingly attentions due to its easy reproducibility and the possibility to greatly reduce the cost. However, significant improvements on the specificity, sensitivity, stability and simplicity of the whole-cell biosensor are still needed prior to its eventual commercialization. Sponsored by US Department of Energy undermore » the contract agreement DE-FG02-07ER64410, we applied the special synthetic biology and directed evolution strategies to improve the effectiveness and performance of whole-cell biosensors. We have constructed different whole-cell biosensors for the mercuric ion and methylmercury detection with metalloregulator MerR, fluorescent protein mCherry and organomercurial lyase MerB. By introducing the mercuric transporter MerT, we were able to increase the detection sensitivity of whole-cell biosensors by at least one fold. By introducing the bio-amplification genetic circuit based on the gene cascade expression system of PRM-cI from bacteriophage l and Pm-XylS2 from Pseudomonas putida, we have increased the detection sensitivity of whole-cell biosensors by 1~2 folds in our tested conditions. With the directed evolution of MerR and subsequent high-throughput screening via color assay and microplate screening, we have dramatically increased the detection sensitivity by up to 10 folds at low concentration of mercury (II) of 1-10nM. Structural modeling and computational analysis of the mutated MerR showed that many mutations could cause the change of a loop to helix, which could be responsible for the increased mercury sensitivity.« less

Development of a microplate coagulation assay for Factor V in human plasma.

PubMed

Tilley, Derek; Levit, Irina; Samis, John A

2011-06-28

Factor V (FV) in its activated form, FVa, is a critical regulator of thrombin generation during fibrin clot formation. There is a need of a simple, fast, and inexpensive microplate-based coagulation assay to measure the functional activity of FV in human plasma. The objective of this study was to develop a microplate-based assay that measures FV coagulation activity during clot formation in human plasma, which is currently not available. The FV assay requires a kinetic microplate reader to measure the change in absorbance at 405nm during fibrin formation in human plasma. The FV assay accurately measures the time, initial rate, and extent of fibrin clot formation in human plasma. The FV microplate assay is simple, fast, economical, sensitive to approx 24-80pM, and multiple samples may be analyzed simultaneously. All the required materials are commercially available. Standard curves of time or initial rate of fibrin clot formation vs FV activity in the 1-stage assay (Without activation by thrombin) may be used to measure FV activity in samples of human plasma. The assay was used to demonstrate that in nine patients with disseminated intravascular coagulation (DIC), the FV 1-stage, 2-stage (With activation by thrombin), and total (2-stage activity - 1-stage activity) activities were decreased, on average, by approximately 54%, 44%, and 42%, respectively, from prolonged clot times when compared to normal pooled human reference plasma (NHP). The results indicate that the FV in the DIC patient plasmas supported both a delayed and slower rate of fibrin clot formation compared with NHP; however, the extent of fibrin clot formation in the DIC patients remained largely unchanged from that observed with NHP. The FV microplate assay may be easily adapted to measure the activity of any coagulation factor using the appropriate factor-deficient plasma and clot initiating reagent. The microplate assay will find use in both research and clinical laboratories to provide measurement

Development of a microplate coagulation assay for Factor V in human plasma

PubMed Central

2011-01-01

Background Factor V (FV) in its activated form, FVa, is a critical regulator of thrombin generation during fibrin clot formation. There is a need of a simple, fast, and inexpensive microplate-based coagulation assay to measure the functional activity of FV in human plasma. The objective of this study was to develop a microplate-based assay that measures FV coagulation activity during clot formation in human plasma, which is currently not available. Methods The FV assay requires a kinetic microplate reader to measure the change in absorbance at 405nm during fibrin formation in human plasma. The FV assay accurately measures the time, initial rate, and extent of fibrin clot formation in human plasma. Results The FV microplate assay is simple, fast, economical, sensitive to approx 24-80pM, and multiple samples may be analyzed simultaneously. All the required materials are commercially available. Standard curves of time or initial rate of fibrin clot formation vs FV activity in the 1-stage assay (Without activation by thrombin) may be used to measure FV activity in samples of human plasma. The assay was used to demonstrate that in nine patients with disseminated intravascular coagulation (DIC), the FV 1-stage, 2-stage (With activation by thrombin), and total (2-stage activity - 1-stage activity) activities were decreased, on average, by approximately 54%, 44%, and 42%, respectively, from prolonged clot times when compared to normal pooled human reference plasma (NHP). The results indicate that the FV in the DIC patient plasmas supported both a delayed and slower rate of fibrin clot formation compared with NHP; however, the extent of fibrin clot formation in the DIC patients remained largely unchanged from that observed with NHP. Conclusions The FV microplate assay may be easily adapted to measure the activity of any coagulation factor using the appropriate factor-deficient plasma and clot initiating reagent. The microplate assay will find use in both research and

Application of a fluorometric microplate algal toxicity assay for riverine periphytic algal species.

PubMed

Nagai, Takashi; Taya, Kiyoshi; Annoh, Hirochica; Ishihara, Satoru

2013-08-01

Although riverine periphytic algae attached to riverbed gravel are dominant species in flowing rivers, there is limited toxicity data on them because of the difficulty in cell culture and assays. Moreover, it is well known that sensitivity to pesticides differ markedly among species, and therefore the toxicity data for multiple species need to be efficiently obtained. In this study, we investigated the use of fluorometric microplate toxicity assay for testing periphytic algal species. We selected five candidate test algal species Desmodesmus subspicatus, Achnanthidium minutissimum, Navicula pelliculosa, Nitzschia palea, and Pseudanabaena galeata. The selected species are dominant in the river, include a wide range of taxon, and represent actual species composition. Other additional species were also used to compare the sensitivity and suitability of the microplate assay. A 96-well microplate was used as a test chamber and algal growth was measured by in-vivo fluorescence. Assay conditions using microplate and fluorometric measurement were established, and sensitivities of 3,5-dichlorophenol as a reference substance were assayed. The 50 percent effect concentrations (EC50s) obtained by fluorometric microplate assay and those obtained by conventional Erlenmeyer flask assay conducted in this study were consistent. Moreover, the EC50 values of 3,5-dichlorophenol were within the reported confidence intervals in literature. These results supported the validity of our microplate assay. Species sensitivity distribution (SSD) analysis was conducted using the EC50s of five species. The SSD was found to be similar to the SSD obtained using additional tested species, suggesting that SSD using the five species largely represents algal sensitivity. Our results provide a useful and efficient method for high-tier probabilistic ecological risk assessment of pesticides. Copyright © 2013 Elsevier Inc. All rights reserved.

Rigid and non-rigid micro-plates: Philippines and Myanmar-Andaman case studies

NASA Astrophysics Data System (ADS)

Rangin, Claude

2016-01-01

Generally, tectonic plates are considered as rigid. Oblique plate convergence favors the development of micro-plates along the converging boundaries. The north-south-trending Philippines archipelago (here named Philippine Mobile Belt, PMB), a few hundreds kilometers wide, is one of such complex tectonic zones. We show here that it is composed of rigid rotating crustal blocks (here called platelets). In Myanmar, the northernmost tip of the Sumatra-Andaman subduction system is another complex zone made of various crustal blocks in-between convergent plates. Yet, contrary to PMB, it sustains internal deformation with platelet buckling, altogether indicative of a non-rigid behavior. Therefore, the two case studies, Philippine Mobile Belt and Myanmar-Andaman micro-plate (MAS), illustrate the complexity of micro-plate tectonics and kinematics at convergent plate boundaries.

Recent advances in merging photonic crystals and plasmonics for bioanalytical applications.

PubMed

Liu, Bing; Monshat, Hosein; Gu, Zhongze; Lu, Meng; Zhao, Xiangwei

2018-05-29

Photonic crystals (PhCs) and plasmonic nanostructures offer the unprecedented capability to control the interaction of light and biomolecules at the nanoscale. Based on PhC and plasmonic phenomena, a variety of analytical techniques have been demonstrated and successfully implemented in many fields, such as biological sciences, clinical diagnosis, drug discovery, and environmental monitoring. During the past decades, PhC and plasmonic technologies have progressed in parallel with their pros and cons. The merging of photonic crystals with plasmonics will significantly improve biosensor performances and enlarge the linear detection range of analytical targets. Here, we review the state-of-the-art biosensors that combine PhC and plasmonic nanomaterials for quantitative analysis. The optical mechanisms of PhCs, plasmonic crystals, and metal nanoparticles (NPs) are presented, along with their integration and potential applications. By explaining the optical coupling of photonic crystals and plasmonics, the review manifests how PhC-plasmonic hybrid biosensors can achieve the advantages, including high sensitivity, low cost, and short assay time as well. The review also discusses the challenges and future opportunities in this fascinating field.

High-throughput microplate technique for enzymatic hydrolysis of lignocellulosic biomass.

PubMed

Chundawat, Shishir P S; Balan, Venkatesh; Dale, Bruce E

2008-04-15

Several factors will influence the viability of a biochemical platform for manufacturing lignocellulosic based fuels and chemicals, for example, genetically engineering energy crops, reducing pre-treatment severity, and minimizing enzyme loading. Past research on biomass conversion has focused largely on acid based pre-treatment technologies that fractionate lignin and hemicellulose from cellulose. However, for alkaline based (e.g., AFEX) and other lower severity pre-treatments it becomes critical to co-hydrolyze cellulose and hemicellulose using an optimized enzyme cocktail. Lignocellulosics are appropriate substrates to assess hydrolytic activity of enzyme mixtures compared to conventional unrealistic substrates (e.g., filter paper, chromogenic, and fluorigenic compounds) for studying synergistic hydrolysis. However, there are few, if any, high-throughput lignocellulosic digestibility analytical platforms for optimizing biomass conversion. The 96-well Biomass Conversion Research Lab (BCRL) microplate method is a high-throughput assay to study digestibility of lignocellulosic biomass as a function of biomass composition, pre-treatment severity, and enzyme composition. The most suitable method for delivering milled biomass to the microplate was through multi-pipetting slurry suspensions. A rapid bio-enzymatic, spectrophotometric assay was used to determine fermentable sugars. The entire procedure was automated using a robotic pipetting workstation. Several parameters that affect hydrolysis in the microplate were studied and optimized (i.e., particle size reduction, slurry solids concentration, glucan loading, mass transfer issues, and time period for hydrolysis). The microplate method was optimized for crystalline cellulose (Avicel) and ammonia fiber expansion (AFEX) pre-treated corn stover. Copyright 2008 Wiley Periodicals, Inc.

A simple technique to reduce evaporation of crystallization droplets by using plate lids with apertures for adding liquids

PubMed Central

Zipper, Lauren E.; Aristide, Xavier; Bishop, Dylan P.; Joshi, Ishita; Kharzeev, Julia; Patel, Krishna B.; Santiago, Brianna M.; Joshi, Karan; Dorsinvil, Kahille; Sweet, Robert M.; Soares, Alexei S.

2014-01-01

A method is described for using plate lids to reduce evaporation in low-volume vapor-diffusion crystallization experiments. The plate lids contain apertures through which the protein and precipitants were added to different crystallization microplates (the reservoir was filled before fitting the lids). Plate lids were designed for each of these commonly used crystallization microplates. This system minimizes the dehydration of crystallization droplets containing just a few nanolitres of protein and precipitant, and results in more reproducible diffraction from the crystals. For each lid design, changes in the weight of the plates were used to deduce the rate of evaporation under different conditions of temperature, air movement, droplet size and precipitant. For comparison, the state of dehydration was also visually assessed throughout the experiment. Finally, X-ray diffraction methods were used to compare the diffraction of protein crystals that were conventionally prepared against those that were prepared on plates with plate lids. The measurements revealed that the plate lids reduced the rate of evaporation by 63–82%. Crystals grown in 5 nl drops that were set up with plate lids diffracted to higher resolution than similar crystals from drops that were set up without plate lids. The results demonstrate that plate lids can be instrumental for improving few-nanolitre crystallizations. PMID:25484231

A simple technique to reduce evaporation of crystallization droplets by using plate lids with apertures for adding liquids.

PubMed

Zipper, Lauren E; Aristide, Xavier; Bishop, Dylan P; Joshi, Ishita; Kharzeev, Julia; Patel, Krishna B; Santiago, Brianna M; Joshi, Karan; Dorsinvil, Kahille; Sweet, Robert M; Soares, Alexei S

2014-12-01

A method is described for using plate lids to reduce evaporation in low-volume vapor-diffusion crystallization experiments. The plate lids contain apertures through which the protein and precipitants were added to different crystallization microplates (the reservoir was filled before fitting the lids). Plate lids were designed for each of these commonly used crystallization microplates. This system minimizes the dehydration of crystallization droplets containing just a few nanolitres of protein and precipitant, and results in more reproducible diffraction from the crystals. For each lid design, changes in the weight of the plates were used to deduce the rate of evaporation under different conditions of temperature, air movement, droplet size and precipitant. For comparison, the state of dehydration was also visually assessed throughout the experiment. Finally, X-ray diffraction methods were used to compare the diffraction of protein crystals that were conventionally prepared against those that were prepared on plates with plate lids. The measurements revealed that the plate lids reduced the rate of evaporation by 63-82%. Crystals grown in 5 nl drops that were set up with plate lids diffracted to higher resolution than similar crystals from drops that were set up without plate lids. The results demonstrate that plate lids can be instrumental for improving few-nanolitre crystallizations.

Quantitative Microplate-Based Respirometry with Correction for Oxygen Diffusion

PubMed Central

2009-01-01

Respirometry using modified cell culture microplates offers an increase in throughput and a decrease in biological material required for each assay. Plate based respirometers are susceptible to a range of diffusion phenomena; as O2 is consumed by the specimen, atmospheric O2 leaks into the measurement volume. Oxygen also dissolves in and diffuses passively through the polystyrene commonly used as a microplate material. Consequently the walls of such respirometer chambers are not just permeable to O2 but also store substantial amounts of gas. O2 flux between the walls and the measurement volume biases the measured oxygen consumption rate depending on the actual [O2] gradient. We describe a compartment model-based correction algorithm to deconvolute the biological oxygen consumption rate from the measured [O2]. We optimize the algorithm to work with the Seahorse XF24 extracellular flux analyzer. The correction algorithm is biologically validated using mouse cortical synaptosomes and liver mitochondria attached to XF24 V7 cell culture microplates, and by comparison to classical Clark electrode oxygraph measurements. The algorithm increases the useful range of oxygen consumption rates, the temporal resolution, and durations of measurements. The algorithm is presented in a general format and is therefore applicable to other respirometer systems. PMID:19555051

Biosensors.

ERIC Educational Resources Information Center

Rechnitz, Garry A.

1988-01-01

Describes theory and principles behind biosensors that incorporate biological components as part of a sensor or probe. Projects major applications in medicine and veterinary medicine, biotechnology, food and agriculture, environmental studies, and the military. Surveys current use of biosensors. (ML)

Enhancing the sensitivity of slow light MZI biosensors through multi-hole defects

NASA Astrophysics Data System (ADS)

Qin, Kun; Zhao, Yiliang; Hu, Shuren; Weiss, Sharon M.

2018-02-01

We demonstrate enhanced detection sensitivity of a slow light Mach-Zehnder interferometer (MZI) sensor by incorporating multi-hole defects (MHDs). Slow light MZI biosensors with a one-dimensional photonic crystal in one arm have been previously shown to improve the performance of traditional MZI sensors based on the increased lightmatter interaction that takes place in the photonic crystal region of the structure. Introducing MHDs in the photonic crystal region increases the available surface area for molecular attachment and further increases the enhanced lightmatter interaction capability of slow light MZIs. The MHDs allow analyte to interact with a greater fraction of the guided wave in the MZI. For a slow light MHD MZI sensor with a 16 μm long sensing arm, a bulk sensitivity of 151,000 rad/RIU-cm is demonstrated experimentally, which is approximately two-fold higher than our previously reported slow light MZI sensors and thirteen-fold higher than traditional MZI biosensors with millimeter length sensing regions. For the label-free detection of nucleic acids, the slow light MZI with MHDs also exhibits a two-fold sensitivity improvement in experiment compared to the slow light MZI without MHDs. Because the detection sensitivity of slow light MHD MZIs scales with the length of the sensing arm, the tradeoff between detection limit and device size can be appropriately mitigated for different applications. All experimental results presented in this work are in good agreement with finite difference-time domain-calculations. Overall, the slow light MZI biosensors with MHDs are a promising platform for highly sensitive and multiplexed lab-on-chip systems.

Nanoscale Biosensor Based on Silicon Photonic Cavity for Home Healthcare Diagnostic Application

NASA Astrophysics Data System (ADS)

Ebrahimy, Mehdi N.; Moghaddam, Aydin B.; Andalib, Alireza; Naziri, Mohammad; Ronagh, Nazli

2015-09-01

In this paper, a new ultra-compact optical biosensor based on photonic crystal (phc) resonant cavity is proposed. This sensor has ability to work in chemical optical processes for the determination and analysis of liquid material. Here, we used an optical filter based on two-dimensional phc resonant cavity on a silicon layer and an active area is created in center of cavity. According to results, with increasing the refractive index of cavity, resonant wavelengths shift so that this phenomenon provides the ability to measure the properties of materials. This novel designed biosensor has more advantage to operate in the biochemical process for example sensing protein and DNA molecule refractive index. This nanoscale biosensor has quality factor higher than 1.5 × 104 and it is suitable to be used in the home healthcare diagnostic applications.

Robust Microplate-Based Methods for Culturing and in Vivo Phenotypic Screening of Chlamydomonas reinhardtii.

PubMed

Haire, Timothy C; Bell, Cody; Cutshaw, Kirstin; Swiger, Brendan; Winkelmann, Kurt; Palmer, Andrew G

2018-01-01

Chlamydomonas reinhardtii (Cr), a unicellular alga, is routinely utilized to study photosynthetic biochemistry, ciliary motility, and cellular reproduction. Its minimal culture requirements, unicellular morphology, and ease of transformation have made it a popular model system. Despite its relatively slow doubling time, compared with many bacteria, it is an ideal eukaryotic system for microplate-based studies utilizing either, or both, absorbance as well as fluorescence assays. Such microplate assays are powerful tools for researchers in the areas of toxicology, pharmacology, chemical genetics, biotechnology, and more. However, while microplate-based assays are valuable tools for screening biological systems, these methodologies can significantly alter the conditions in which the organisms are cultured and their subsequent physiology or morphology. Herein we describe a novel method for the microplate culture and in vivo phenotypic analysis of growth, viability, and photosynthetic pigments of C. reinhardtii . We evaluated the utility of our assay by screening silver nanoparticles for their effects on growth and viability. These methods are amenable to a wide assortment of studies and present a significant advancement in the methodologies available for research involving this model organism.

QCM-nanomagnetic beads biosensor for lead ion detection.

PubMed

Zhang, Qingli; Cui, Haixia; Xiong, Xingliang; Chen, Jun; Wang, Ying; Shen, Jia; Luo, Yiting; Chen, Longcong

2018-01-15

As lead poses a serious threat to humans even in small amounts, all kinds of lead detection sensors with high sensitivity and selectivity are being constantly improved and put forward. In this report, a novel, simple and label-free quartz crystal microbalance (QCM) biosensor is proposed for detecting lead ions (Pb 2+ ). The biosensor takes full advantage of the high specificity of GR-5 DNAzyme to Pb 2+ and the high sensitivity of QCM. In particular, nanomagnetic beads (NMBs) are used as a novel and effective mean of signal amplification in the biosensor because of their mass and their ability to enhance the inductive effect, which are very beneficial for both higher sensitivity and a lower detection limit. In practice, GR-5 DNAzyme, innovatively combined with NMBs, was modified on the gold electrode of the QCM through gold-sulfur self-assembly. When the electrode was exposed to Pb 2+ solution, DNAzyme was severed into two parts at the RNA site (rA), along with the release of NMBs, which caused a great increase in frequency shift of the QCM electrode. Finally, a perfect linear correlation between the logarithm of Pb 2+ concentration and the change in frequency was obtained from 1 pM to 50 nM, with a detection limit as low as 0.3 pM. Moreover, the biosensor shows both an average recovery of 97 ± 6% in a drinking water sample and an excellent specificity for Pb 2+ compared with other metal ions.

Biosensors for Cell Analysis.

PubMed

Zhou, Qing; Son, Kyungjin; Liu, Ying; Revzin, Alexander

2015-01-01

Biosensors first appeared several decades ago to address the need for monitoring physiological parameters such as oxygen or glucose in biological fluids such as blood. More recently, a new wave of biosensors has emerged in order to provide more nuanced and granular information about the composition and function of living cells. Such biosensors exist at the confluence of technology and medicine and often strive to connect cell phenotype or function to physiological or pathophysiological processes. Our review aims to describe some of the key technological aspects of biosensors being developed for cell analysis. The technological aspects covered in our review include biorecognition elements used for biosensor construction, methods for integrating cells with biosensors, approaches to single-cell analysis, and the use of nanostructured biosensors for cell analysis. Our hope is that the spectrum of possibilities for cell analysis described in this review may pique the interest of biomedical scientists and engineers and may spur new collaborations in the area of using biosensors for cell analysis.

A High-Content Assay for Biosensor Validation and for Examining Stimuli that Affect Biosensor Activity.

PubMed

Slattery, Scott D; Hahn, Klaus M

2014-12-01

Biosensors are valuable tools used to monitor many different protein behaviors in vivo. Demand for new biosensors is high, but their development and characterization can be difficult. During biosensor design, it is necessary to evaluate the effects of different biosensor structures on specificity, brightness, and fluorescence responses. By co-expressing the biosensor with upstream proteins that either stimulate or inhibit the activity reported by the biosensor, one can determine the difference between the biosensor's maximally activated and inactivated state, and examine response to specific proteins. We describe here a method for biosensor validation in a 96-well plate format using an automated microscope. This protocol produces dose-response curves, enables efficient examination of many parameters, and unlike cell suspension assays, allows visual inspection (e.g., for cell health and biosensor or regulator localization). Optimization of single-chain and dual-chain Rho GTPase biosensors is addressed, but the assay is applicable to any biosensor that can be expressed or otherwise loaded in adherent cells. The assay can also be used for purposes other than biosensor validation, using a well-characterized biosensor as a readout for effects of upstream molecules. Copyright © 2014 John Wiley & Sons, Inc.

A simple technique to reduce evaporation of crystallization droplets by using plate lids with apertures for adding liquids

DOE PAGES

Zipper, Lauren E.; Aristide, Xavier; Bishop, Dylan P.; ...

2014-11-28

A method is described for using plate lids to reduce evaporation in low-volume vapor-diffusion crystallization experiments. The plate lids contain apertures through which the protein and precipitants were added to different crystallization microplates (the reservoir was filled before fitting the lids). Plate lids were designed for each of these commonly used crystallization microplates. This system minimizes the dehydration of crystallization droplets containing just a few nanolitres of protein and precipitant, and results in more reproducible diffraction from the crystals. For each lid design, changes in the weight of the plates were used to deduce the rate of evaporation under differentmore » conditions of temperature, air movement, droplet size and precipitant. For comparison, the state of dehydration was also visually assessed throughout the experiment. Finally, X-ray diffraction methods were used to compare the diffraction of protein crystals that were conventionally prepared against those that were prepared on plates with plate lids. The measurements revealed that the plate lids reduced the rate of evaporation by 63–82%. Crystals grown in 5 nl drops that were set up with plate lids diffracted to higher resolution than similar crystals from drops that were set up without plate lids. Ultimately, the results demonstrate that plate lids can be instrumental for improving few-nanolitre crystallizations.« less

High-throughput measurements of the optical redox ratio using a commercial microplate reader.

PubMed

Cannon, Taylor M; Shah, Amy T; Walsh, Alex J; Skala, Melissa C

2015-01-01

There is a need for accurate, high-throughput, functional measures to gauge the efficacy of potential drugs in living cells. As an early marker of drug response in cells, cellular metabolism provides an attractive platform for high-throughput drug testing. Optical techniques can noninvasively monitor NADH and FAD, two autofluorescent metabolic coenzymes. The autofluorescent redox ratio, defined as the autofluorescence intensity of NADH divided by that of FAD, quantifies relative rates of cellular glycolysis and oxidative phosphorylation. However, current microscopy methods for redox ratio quantification are time-intensive and low-throughput, limiting their practicality in drug screening. Alternatively, high-throughput commercial microplate readers quickly measure fluorescence intensities for hundreds of wells. This study found that a commercial microplate reader can differentiate the receptor status of breast cancer cell lines (p < 0.05) based on redox ratio measurements without extrinsic contrast agents. Furthermore, microplate reader redox ratio measurements resolve response (p < 0.05) and lack of response (p > 0.05) in cell lines that are responsive and nonresponsive, respectively, to the breast cancer drug trastuzumab. These studies indicate that the microplate readers can be used to measure the redox ratio in a high-throughput manner and are sensitive enough to detect differences in cellular metabolism that are consistent with microscopy results.

Surface stress-based biosensors.

PubMed

Sang, Shengbo; Zhao, Yuan; Zhang, Wendong; Li, Pengwei; Hu, Jie; Li, Gang

2014-01-15

Surface stress-based biosensors, as one kind of label-free biosensors, have attracted lots of attention in the process of information gathering and measurement for the biological, chemical and medical application with the development of technology and society. This kind of biosensors offers many advantages such as short response time (less than milliseconds) and a typical sensitivity at nanogram, picoliter, femtojoule and attomolar level. Furthermore, it simplifies sample preparation and testing procedures. In this work, progress made towards the use of surface stress-based biosensors for achieving better performance is critically reviewed, including our recent achievement, the optimally circular membrane-based biosensors and biosensor array. The further scientific and technological challenges in this field are also summarized. Critical remark and future steps towards the ultimate surface stress-based biosensors are addressed. Copyright © 2013 Elsevier B.V. All rights reserved.

Bio-recognition and detection using liquid crystals.

PubMed

Hussain, A; Pina, A S; Roque, A C A

2009-09-15

Liquid crystals (LCs) are used extensively by the electronics industry as display devices. Advances in the understanding of the liquid crystalline phase and the chemistry therein lead to the development of LC exhibiting faster switching speed with greater twist angle. This in turn lead to the emergence of liquid crystal displays, rendering dial-and-needle based displays (such as those used in various meters) and cathode ray tubes obsolete. In this article, we review the history of LC and their emergence as an invaluable material for display devices and the more recent discovery of their use as sensing elements in biosensors. This new application of LC as tools in the development of fast and simple biosensors is envisaged to gain more importance in the foreseeable future.

Design and process development of a photonic crystal polymer biosensor for point-of-care diagnostics

NASA Astrophysics Data System (ADS)

Dortu, F.; Egger, H.; Kolari, K.; Haatainen, T.; Furjes, P.; Fekete, Z.; Bernier, D.; Sharp, G.; Lahiri, B.; Kurunczi, S.; Sanchez, J.-C.; Turck, N.; Petrik, P.; Patko, D.; Horvath, R.; Eiden, S.; Aalto, T.; Watts, S.; Johnson, N. P.; De La Rue, R. M.; Giannone, D.

2011-07-01

In this work, we report advances in the fabrication and anticipated performance of a polymer biosensor photonic chip developed in the European Union project P3SENS (FP7-ICT4-248304). Due to the low cost requirements of point-ofcare applications, the photonic chip is fabricated from nanocomposite polymeric materials, using highly scalable nanoimprint- lithography (NIL). A suitable microfluidic structure transporting the analyte solutions to the sensor area is also fabricated in polymer and adequately bonded to the photonic chip. We first discuss the design and the simulated performance of a high-Q resonant cavity photonic crystal sensor made of a high refractive index polyimide core waveguide on a low index polymer cladding. We then report the advances in doped and undoped polymer thin film processing and characterization for fabricating the photonic sensor chip. Finally the development of the microfluidic chip is presented in details, including the characterisation of the fluidic behaviour, the technological and material aspects of the 3D polymer structuring and the stable adhesion strategies for bonding the fluidic and the photonic chips, with regards to the constraints imposed by the bioreceptors supposedly already present on the sensors.

Biosensors of bacterial cells.

PubMed

Burlage, Robert S; Tillmann, Joshua

2017-07-01

Biosensors are devices which utilize both an electrical component (transducer) and a biological component to study an environment. They are typically used to examine biological structures, organisms and processes. The field of biosensors has now become so large and varied that the technology can often seem impenetrable. Yet the principles which underlie the technology are uncomplicated, even if the details of the mechanisms are elusive. In this review we confine our analysis to relatively current advancements in biosensors for the detection of whole bacterial cells. This includes biosensors which rely on an added labeled component and biosensors which do not have a labeled component and instead detect the binding event or bound structure on the transducer. Methods to concentrate the bacteria prior to biosensor analysis are also described. The variety of biosensor types and their actual and potential uses are described. Copyright © 2016 Elsevier B.V. All rights reserved.

Robust Microplate-Based Methods for Culturing and in Vivo Phenotypic Screening of Chlamydomonas reinhardtii

PubMed Central

Haire, Timothy C.; Bell, Cody; Cutshaw, Kirstin; Swiger, Brendan; Winkelmann, Kurt; Palmer, Andrew G.

2018-01-01

Chlamydomonas reinhardtii (Cr), a unicellular alga, is routinely utilized to study photosynthetic biochemistry, ciliary motility, and cellular reproduction. Its minimal culture requirements, unicellular morphology, and ease of transformation have made it a popular model system. Despite its relatively slow doubling time, compared with many bacteria, it is an ideal eukaryotic system for microplate-based studies utilizing either, or both, absorbance as well as fluorescence assays. Such microplate assays are powerful tools for researchers in the areas of toxicology, pharmacology, chemical genetics, biotechnology, and more. However, while microplate-based assays are valuable tools for screening biological systems, these methodologies can significantly alter the conditions in which the organisms are cultured and their subsequent physiology or morphology. Herein we describe a novel method for the microplate culture and in vivo phenotypic analysis of growth, viability, and photosynthetic pigments of C. reinhardtii. We evaluated the utility of our assay by screening silver nanoparticles for their effects on growth and viability. These methods are amenable to a wide assortment of studies and present a significant advancement in the methodologies available for research involving this model organism. PMID:29623083

Large-scale preparation of shape controlled SnO and improved capacitance for supercapacitors: from nanoclusters to square microplates

NASA Astrophysics Data System (ADS)

Wang, Lu; Ji, Hongmei; Zhu, Feng; Chen, Zhi; Yang, Yang; Jiang, Xuefan; Pinto, João; Yang, Gang

2013-07-01

Here, we first provide a facile ultrasonic-assisted synthesis of SnO using SnCl2 and the organic solvent of ethanolamine (ETA). The moderate alkalinity of ETA and ultrasound play very important roles in the synthesis of SnO. After the hydrolysis of the intermediate of ETA-Sn(ii), the as-synthesized SnO nanoclusters undergo assembly, amalgamation, and preferential growth to microplates in hydrothermal treatment. The as-synthesized SnO was characterized by scanning electron microscopy (SEM), transmission electron microscopy (TEM), high-resolution transmission electron microscopy (HRTEM), ultraviolet-visible absorption spectroscopy (UV-vis) and X-ray diffraction (XRD). To explore its potential applications in energy storage, SnO was fabricated into a supercapacitor electrode and characterized by cyclic voltammetry (CV), electrochemical impedance spectroscopy (EIS), and galvanostatic charge-discharge measurements. The as-synthesized SnO exhibits remarkable pseudocapacitive activity including high specific capacitance (208.9 F g-1 at 0.1 A g-1), good rate capability (65.8 F g-1 at 40 A g-1), and excellent cycling stability (retention 119.3% after 10 000 cycles) for application in supercapacitors. The capacitive behavior of SnO with various crystal morphologies was observed by fitted EIS using an equivalent circuit. The novel synthetic route for SnO is a convenient and potential way to large-scale production of microplates which is expected to be applicable in the synthesis of other metal oxide nanoparticles.Here, we first provide a facile ultrasonic-assisted synthesis of SnO using SnCl2 and the organic solvent of ethanolamine (ETA). The moderate alkalinity of ETA and ultrasound play very important roles in the synthesis of SnO. After the hydrolysis of the intermediate of ETA-Sn(ii), the as-synthesized SnO nanoclusters undergo assembly, amalgamation, and preferential growth to microplates in hydrothermal treatment. The as-synthesized SnO was characterized by scanning

Establishment and validation of a method for multi-dose irradiation of cells in 96-well microplates

DOE Office of Scientific and Technical Information (OSTI.GOV)

Abatzoglou, Ioannis; Zois, Christos E.; Pouliliou, Stamatia

2013-02-15

Highlights: ► We established a method for multi-dose irradiation of cell cultures within a 96-well plate. ► Equations to adjust to preferable dose levels are produced and provided. ► Up to eight different dose levels can be tested in one microplate. ► This method results in fast and reliable estimation of radiation dose–response curves. -- Abstract: Microplates are useful tools in chemistry, biotechnology and molecular biology. In radiobiology research, these can be also applied to assess the effect of a certain radiation dose delivered to the whole microplate, to test radio-sensitivity, radio-sensitization or radio-protection. Whether different radiation doses can bemore » accurately applied to a single 96-well plate to further facilitate and accelerated research by one hand and spare funds on the other, is a question dealt in the current paper. Following repeated ion-chamber, TLD and radiotherapy planning dosimetry we established a method for multi-dose irradiation of cell cultures within a 96-well plate, which allows an accurate delivery of desired doses in sequential columns of the microplate. Up to eight different dose levels can be tested in one microplate. This method results in fast and reliable estimation of radiation dose–response curves.« less

Cholinesterase-based biosensors.

PubMed

Štěpánková, Šárka; Vorčáková, Katarína

2016-01-01

Recently, cholinesterase-based biosensors are widely used for assaying anticholinergic compounds. Primarily biosensors based on enzyme inhibition are useful analytical tools for fast screening of inhibitors, such as organophosphates and carbamates. The present review is aimed at compilation of the most important facts about cholinesterase based biosensors, types of physico-chemical transduction, immobilization strategies and practical applications.

Modified microplate method for rapid and efficient estimation of siderophore produced by bacteria.

PubMed

Arora, Naveen Kumar; Verma, Maya

2017-12-01

In this study, siderophore production by various bacteria amongst the plant-growth-promoting rhizobacteria was quantified by a rapid and efficient method. In total, 23 siderophore-producing bacterial isolates/strains were taken to estimate their siderophore-producing ability by the standard method (chrome azurol sulphonate assay) as well as 96 well microplate method. Production of siderophore was estimated in percent siderophore unit by both the methods. It was observed that data obtained by both methods correlated positively with each other proving the correctness of microplate method. By the modified microplate method, siderophore production by several bacterial strains can be estimated both qualitatively and quantitatively at one go, saving time, chemicals, making it very less tedious, and also being cheaper in comparison with the method currently in use. The modified microtiter plate method as proposed here makes it far easier to screen the plant-growth-promoting character of plant-associated bacteria.

A microplate assay for measuring cell death in C2C12 cells.

PubMed

Lima, Tanes; Silveira, Leonardo

2018-03-22

The main goal of this study was to develop a straightforward and rapid microplate assay for measuring propidium iodide (PI) in C2C12 cells. The PI method proves to be an efficient quantitative assay for analyzing cell viability through PI fluorescence analysis. Importantly, the protocol takes less than 30 minutes, and the results are reproducible. C2C12 cells were exposed to an increasing concentration of palmitate for a period of 24 hours to induce cell death, and the PI fluorescence increased in a concentration-dependent manner. Evaluation of mitochondrial function and reactive oxygen species production validated the deleterious effects of palmitate treatment. Also, the microplate PI assay demonstrated high sensitivity as indicated by the detection of modest fluctuations in cell viability in response to catalase overexpression in palmitate-treated cells. The microplate PI assay, therefore, offers an accurate method to be used for in vitro studies.

Biosensor development.

PubMed

Ziegler, C; Göpel, W

1998-10-01

Current biosensor developments can be summarised by different trends. For traditional enzymatic biosensors such as glucose sensors, steady improvements of well known basic principles have been made in order to achieve better sensor stability. On the other hand, new affinity sensors such as nucleic acid sensors, transmembrane sensors, and sensors utilising whole cells or even cell networks have become of increasing interest. New ways to miniaturise biosensors and to control their interfaces down to the molecular level have been introduced (the bioelectronics approach). High-throughput screening based on various signal transduction principles has become of increasing importance.

Use of a Parasitic Wasp as a Biosensor

PubMed Central

Olson, Dawn; Rains, Glen

2014-01-01

Screening cargo for illicit substances is in need of rapid high-throughput inspection systems that accurately identify suspicious cargo. Here we investigate the ability of a parasitic wasp, Microplitis croceipes to detect and respond to methyl benzoate, the volatile component of cocaine, by examining their response to training concentrations, their sensitivity at low concentrations, and their ability to detect methyl benzoate when two concealment substances (green tea and ground coffee) are added to the testing arena. Utilizing classical associative learning techniques with sucrose as reward, we found that M. croceipes learns individual concentrations of methyl benzoate, and they can generalize this learning to concentrations 100× lower than the training concentration. Their sensitivity to methyl benzoate is very low at an estimated 3 ppb. They are also able to detect methyl benzoate when covered completely by green tea, but were not able to detect methyl benzoate when covered completely by coffee grounds. Habituation to the tea and coffee odors prior to testing improves their responses, resulting in effective detection of methyl benzoate covered by the coffee grounds. With the aid of the portable device called ‘the wasp hound’, the wasps appear to have potential to be effective on-site biosensors for the detection of cocaine. PMID:25587415

Photoelectrochemical enzymatic biosensors.

PubMed

Zhao, Wei-Wei; Xu, Jing-Juan; Chen, Hong-Yuan

2017-06-15

Enzymatic biosensors have been valuable bioanalytical devices for analysis of diverse targets in disease diagnosis, biological and biomedical research, etc. Photoelectrochemical (PEC) bioanalysis is a recently emerged method that promptly becoming a subject of new research interests due to its attractive potential for future bioanalysis with high sensitivity and specificity. PEC enzymatic biosensors integrate the inherent sensitivities of PEC bioanalysis and the selectivity of enzymes and thus share their both advantages. Currently, PEC enzymatic biosensors have become a hot topic of significant research and the recent impetus has grown rapidly as demonstrated by increased research papers. Given the pace of advances in this area, this review will make a thorough discussion and survey on the fundamentals, sensing strategies, applications and the state of the art in PEC enzymatic biosensors, followed by future prospects based on our own opinions. We hope this work could provide an accessible introduction to PEC enzymatic biosensors for any scientist. Copyright © 2016 Elsevier B.V. All rights reserved.

Slotted Photonic Crystal Sensors

PubMed Central

Scullion, Mark G.; Krauss, Thomas F.; Di Falco, Andrea

2013-01-01

Optical biosensors are increasingly being considered for lab-on-a-chip applications due to their benefits such as small size, biocompatibility, passive behaviour and lack of the need for fluorescent labels. The light guiding mechanisms used by many of them results in poor overlap of the optical field with the target molecules, reducing the maximum sensitivity achievable. This review article presents a new platform for optical biosensors, namely slotted photonic crystals, which provide higher sensitivities due to their ability to confine, spatially and temporally, the optical mode peak within the analyte itself. Loss measurements showed values comparable to standard photonic crystals, confirming their ability to be used in real devices. A novel resonant coupler was designed, simulated, and experimentally tested, and was found to perform better than other solutions within the literature. Combining with cavities, microfluidics and biological functionalization allowed proof-of-principle demonstrations of protein binding to be carried out. Higher sensitivities were observed in smaller structures than possible with most competing devices reported in the literature. This body of work presents slotted photonic crystals as a realistic platform for complete on-chip biosensing; addressing key design, performance and application issues, whilst also opening up exciting new ideas for future study. PMID:23503295

Biosensor commercialization strategy - a theoretical approach.

PubMed

Lin, Chin-Tsai; Wang, Su-Man

2005-01-01

Biosensors are analytical devices, which use biological interactions to provide either qualitative or quantitative results. They are extensively employed in many fields such as clinical diagnosis and biomedicine, military applications, anti-terrorism, farm, garden and veterinary analysis, process control, fermentation control and analysis, pharmaceutical and drug analysis, food and drink production and analysis, pollution control and monitoring, microbiology, bacterial and viral analysis, mining, and industrial and toxic gases. The biosensor market has significantly increased and will be mushrooming in the next decade. The total biosensor market is estimated to be 10.8 billion dollars by 2007. The emerging biosensor market presents both opportunities and obstacles to start-up biosensor entrepreneurs. The major challenge and threat for these entrepreneurs is how to predict the biosensor market and how to convert promising biosensor technology into commercialized biosensors. By adopting a simple commercialization strategy framework, we identify two key elements of biosensor commercialization strategy: excludability and complementary asset. We further divide biosensor commercialization environments into four distinct sub-environments: the Attacker's Advantage, Reputation-Based Idea Trading, Greenfield Competition and Ideas Factories. This paper explains how the interaction between these two key elements shapes biosensor commercialization strategy and biosensor industry dynamics. This paper also discusses alternative commercialization strategies for each specific commercialization environment and how to choose from these alternatives. The analysis of this study further provides a good reference for start-up biosensor entrepreneurs to formulate effective biosensor commercialization strategy.

Label-free liquid crystal biosensor based on specific oligonucleotide probes for heavy metal ions.

PubMed

Yang, Shengyuan; Wu, Chao; Tan, Hui; Wu, Yan; Liao, Shuzhen; Wu, Zhaoyang; Shen, Guoli; Yu, Ruqin

2013-01-02

In this study, to enhance the capability of metal ions disturbing the orientation of liquid crystals (LCs), we designed a new label-free LC biosensor for the highly selective and sensitive detection of heavy metal ions. This strategy makes use of the target-induced DNA conformational change to enhance the disruption of target molecules for the orientation of LC leading to an amplified optical signal. The Hg(2+) ion, which possesses a unique property to bind specifically to two DNA thymine (T) bases, is used as a model heavy metal ion. In the presence of Hg(2+), the specific oligonucleotide probes form a conformational reorganization of the oligonucleotide probes from hairpin structure to duplex-like complexes. The duplex-like complexes are then bound on the triethoxysilylbutyraldehyde/N,N-dimethyl-N-octadecyl (3-aminopropyl) trimethoxysilyl chloride (TEA/DMOAP)-coated substrate modified with capture probes, which can greatly distort the orientational profile of LC, making the optical image of LC cell birefringent as a result. The optical signal of LC sensor has a visible change at the Hg(2+) concentration of low to 0.1 nM, showing good detection sensitivity. The cost-effective LC sensing method can translate the concentration signal of heavy metal ions in solution into the presence of DNA duplexes and is expected to be a sensitive detection platform for heavy metal ions and other small molecule monitors.

Determination of Microbial Extracellular Enzyme Activity in Waters, Soils, and Sediments using High Throughput Microplate Assays

PubMed Central

Jackson, Colin R.; Tyler, Heather L.; Millar, Justin J.

2013-01-01

Much of the nutrient cycling and carbon processing in natural environments occurs through the activity of extracellular enzymes released by microorganisms. Thus, measurement of the activity of these extracellular enzymes can give insights into the rates of ecosystem level processes, such as organic matter decomposition or nitrogen and phosphorus mineralization. Assays of extracellular enzyme activity in environmental samples typically involve exposing the samples to artificial colorimetric or fluorometric substrates and tracking the rate of substrate hydrolysis. Here we describe microplate based methods for these procedures that allow the analysis of large numbers of samples within a short time frame. Samples are allowed to react with artificial substrates within 96-well microplates or deep well microplate blocks, and enzyme activity is subsequently determined by absorption or fluorescence of the resulting end product using a typical microplate reader or fluorometer. Such high throughput procedures not only facilitate comparisons between spatially separate sites or ecosystems, but also substantially reduce the cost of such assays by reducing overall reagent volumes needed per sample. PMID:24121617

Determination of microbial extracellular enzyme activity in waters, soils, and sediments using high throughput microplate assays.

PubMed

Jackson, Colin R; Tyler, Heather L; Millar, Justin J

2013-10-01

Much of the nutrient cycling and carbon processing in natural environments occurs through the activity of extracellular enzymes released by microorganisms. Thus, measurement of the activity of these extracellular enzymes can give insights into the rates of ecosystem level processes, such as organic matter decomposition or nitrogen and phosphorus mineralization. Assays of extracellular enzyme activity in environmental samples typically involve exposing the samples to artificial colorimetric or fluorometric substrates and tracking the rate of substrate hydrolysis. Here we describe microplate based methods for these procedures that allow the analysis of large numbers of samples within a short time frame. Samples are allowed to react with artificial substrates within 96-well microplates or deep well microplate blocks, and enzyme activity is subsequently determined by absorption or fluorescence of the resulting end product using a typical microplate reader or fluorometer. Such high throughput procedures not only facilitate comparisons between spatially separate sites or ecosystems, but also substantially reduce the cost of such assays by reducing overall reagent volumes needed per sample.

DNA nanotechnology-enabled biosensors.

PubMed

Chao, Jie; Zhu, Dan; Zhang, Yinan; Wang, Lianhui; Fan, Chunhai

2016-02-15

Biosensors employ biological molecules to recognize the target and utilize output elements which can translate the biorecognition event into electrical, optical or mass-sensitive signals to determine the quantities of the target. DNA-based biosensors, as a sub-field to biosensor, utilize DNA strands with short oligonucleotides as probes for target recognition. Although DNA-based biosensors have offered a promising alternative for fast, simple and cheap detection of target molecules, there still exist key challenges including poor stability and reproducibility that hinder their competition with the current gold standard for DNA assays. By exploiting the self-recognition properties of DNA molecules, researchers have dedicated to make versatile DNA nanostructures in a highly rigid, controllable and functionalized manner, which offers unprecedented opportunities for developing DNA-based biosensors. In this review, we will briefly introduce the recent advances on design and fabrication of static and dynamic DNA nanostructures, and summarize their applications for fabrication and functionalization of DNA-based biosensors. Copyright © 2015 Elsevier B.V. All rights reserved.

Electrochemical biosensors for hormone analyses.

PubMed

Bahadır, Elif Burcu; Sezgintürk, Mustafa Kemal

2015-06-15

Electrochemical biosensors have a unique place in determination of hormones due to simplicity, sensitivity, portability and ease of operation. Unlike chromatographic techniques, electrochemical techniques used do not require pre-treatment. Electrochemical biosensors are based on amperometric, potentiometric, impedimetric, and conductometric principle. Amperometric technique is a commonly used one. Although electrochemical biosensors offer a great selectivity and sensitivity for early clinical analysis, the poor reproducible results, difficult regeneration steps remain primary challenges to the commercialization of these biosensors. This review summarizes electrochemical (amperometric, potentiometric, impedimetric and conductometric) biosensors for hormone detection for the first time in the literature. After a brief description of the hormones, the immobilization steps and analytical performance of these biosensors are summarized. Linear ranges, LODs, reproducibilities, regenerations of developed biosensors are compared. Future outlooks in this area are also discussed. Copyright © 2014 Elsevier B.V. All rights reserved.

Cryoalgotox: Use of cryopreserved alga in a semistatic microplate test

DOE Office of Scientific and Technical Information (OSTI.GOV)

Benhra, A.; Radetski, C.M.; Ferard, J.F.

1997-03-01

Use of cryopreserved alga Selenastrum capricornutum has been evaluated as a simple and cost-efficient procedure in a new semistatic algal ecotoxicity test. Experiments have been conducted to compare performance criteria of this method, named Cryoalgotox, versus the classic microplate test using fresh algae. Cryoalgotox 72-h 50% effective concentrations (EC50s) determined with Cd{sup 2+}, Cu{sup 2+}, Cr{sup 6+}, and atrazine were more sensitive, repeatable (low coefficients of variation), and reproducible (low time effect) than the results obtained with the classical microplate tests. The effect of storage time at {minus}80 C on the sensitivity of the algae was assessed using cadmium asmore » a toxic reference; it was shown that algae stored at {minus}80 C over a 3-month period gave comparable toxicity results to those found with fresh algae.« less

A microplate reader-based method to quantify NADH-cytochrome b5 reductase activity for diagnosis of recessive congenital methaemoglobinemia.

PubMed

Kedar, Prabhakar; Desai, Anand; Warang, Prashant; Colah, Roshan

2017-05-01

Congenital methemoglobinemia due to NADH-cytochrome b5 reductase 3 (CYB5R3) deficiencies is an autosomal recessive disorder that occurs sporadically worldwide, A sensitive, accurate, and rapid analysis of NADH-CYB5R enzyme concentrations is necessary for the diagnosis of RCM. Here we present an alternative microplate method that is based on a standard 96-well microplate format and microplate reader that simplify the quantification of NADH-CYB5R activity. TECAN (Infinite 200 PRO series) microplate reader with Tecan's proven Magellan™ software measured the NADH-CYB5R enzyme activity in 250 normal controls and previously diagnosed 25 cases of RCM due to NADH-CYB5R deficiency in the Indian population using 96-well microplates using 200 μl of total reaction mixture and also compared with standard spectrophotometric assay. We have also studied stability of the hemolysate stored at 4 and -20°C temperature. Enzyme activity in all 25 samples ranged from 6.09 to 10.07 IU/g Hb (mean ± SD: 8.08 ± 1.99 IU/g Hb) where as normal control ranged (n = 250) between 13.42 and 21.58 IU/g Hb) (mean ± SD: 17.5 ± 4.08 IU/g of Hb). Data obtained from the microplate reader were compared with standard spectrophotometer method and found 100% concordance using both methods. Microplate method allows differentiating between normal, deficient and intermediate enzyme activity. It was observed that samples had significant loss of activity when stored at 4°C and retained stable activity at -20°C for 1 week time. Our new method, incorporating a whole process of enzyme assay into a microplate format is readily applicable and allows rapid monitoring of enzyme assay. It is readily applicable to quantitative assay on pediatric sample as well as large number of samples for population screening.

Functional and radiologic outcome of open reduction and internal fixation of condylar head and neck fractures using miniplate or microplate system.

PubMed

Xie, Si-Tian; Singhal, Dhruv; Chen, Chien-Tzung; Chen, Yu-Ray

2013-12-01

Although the appropriate management of condylar process fractures after miniplate or microplate fixation has been described, there has been no comparative analysis of these plating systems. A retrospective review of patients who underwent open reduction and internal fixation (ORIF) of condylar head or neck fractures at our institution from January 2000 through August 2010 identified 70 patients. Of these, 38 were treated with microplates and 32 with miniplates. The primary functional and radiographic results were the maximal mouth opening and condylar bone resorption, respectively. The rates of complications, including malocclusion, chin deviation, temporomandibular joint complaints, and facial nerve palsy, were recorded. The maximal mouth opening was larger in the microplate group than in the miniplate group throughout the follow-up period; this difference was statistically significant 12 (P = 0.020), 18 (P = 0.026), and 24 (P = 0.032) months after ORIF. Similarly, the radiographic scores for bone resorption and condyle morphology were significantly better in the microplate group than in the miniplate group throughout the follow-up period [6 (P = 0.011), 12 (P = 0.035), 24 (P = 0.026), and 48 (P = 0.040) months after ORIF]. Moreover, patients who underwent miniplate fixation experienced a significantly higher incidence of temporomandibular joint click than those who underwent microplate fixation (P = 0.014). Microplates limit dissection, providing excellent fixation for intracapsular condylar head fractures, and also provide adequate rigidity for fixation of condylar neck fractures. Microplate fixation of condylar head and neck fractures yielded excellent functional and radiographic results. The rates of complications after microplate fixation were equal to or less than those in the miniplate group. Prospective studies are needed to confirm these findings.

Magnetization measurements of Sr2RuO4-Ru eutectic microplates using dc-SQUIDs

NASA Astrophysics Data System (ADS)

Nago, Y.; Sakuma, D.; Ishiguro, R.; Kashiwaya, S.; Nomura, S.; Kono, K.; Maeno, Y.; Takayanagi, H.

2018-03-01

We report magnetization measurements of Sr2RuO4-Ru eutectic microplates using micro-dc-SQUIDs. Sr2RuO4 is considered as a chiral p-wave superconductor and hence Sr2RuO4-Ru eutectic becomes in an unstable state with a superconducting phase frustration between a chiral p-wave state of Sr2RuO4 and a s-wave state of Ru. To compensate the frustration, a single quantum vortex is spontaneously formed at the center of the Ru inclusion at sufficiently low temperatures. However, such a spontaneous vortex state has not been experimentally observed yet. In this study, we prepared a micro-dc-SQUID and a Sr2RuO4-Ru eutectic microplate containing a single Ru-inclusion at the center of the microplate. We performed magnetization measurements down below the superconducting transition temperature of the Ru inclusion to investigate the spontaneous Ru-center vortex state.

Platelet aggregation inhibitors from Philippine marine invertebrate samples screened in a new microplate assay.

PubMed

Pimentel, Sheila Marie V; Bojo, Zenaida P; Roberto, Amy V D; Lazaro, Jose Enrico H; Mangalindan, Gina C; Florentino, Leila M; Lim-Navarro, Pilar; Tasdemir, Deniz; Ireland, Chris M; Concepcion, Gisela P

2003-01-01

A new microplate assay for Ca(2+)-induced platelet aggregation as detected by Giemsa dye was used to screen marine invertebrate samples from the Philippines for inhibitors of human platelet aggregation. Out of 261 crude methanol extracts of marine sponges and tunicates, 25 inhibited aggregation at 2 mg/ml. Inhibition of agonist-induced aggregation in an aggregometer was used to confirm results of the microplate assay and to determine the specific mode of inhibition of 2 samples. The marine sponge Xestospongia sp. yielded a xestospongin/araguspongine-type molecule that inhibited collagen-induced aggregation by 87% at 2 micro g/ml, and epinephrine-induced aggregation by 78% at 20 micro g/ml, while the marine sponge Aplysina sp. yielded 5,6-dibromotryptamine, which inhibited epinephrine-induced aggregation by 51% at 20 micro g/ml. In this study we have found that the microplate assay is a simple, inexpensive, yet useful preliminary tool to qualitatively screen a large number of marine samples for antiplatelet aggregation activity.

Influence of electrical double-layer dispersion forces and size dependency on pull-in instability of clamped microplate immersed in ionic liquid electrolytes

NASA Astrophysics Data System (ADS)

Karimipour, I.; Beni, Yaghoub Tadi; Taheri, N.

2017-10-01

Plate-type clamped microplate is of the most common constructive elements for developing in-liquid-operating devices. While the electromechanical behavior of clamped microplate in non-liquid environments has exclusively been addressed in the literature, no theoretical studies have yet been conducted on precise modeling of the clamped microplate in electrolyte liquid. Herein, the electromechanical response and instability of the clamped microplate immersed in ionic electrolyte media are investigated. The electrochemical force field is determined using double layer theory and linearized Poisson-Boltzmann equation. The presence of dispersion forces, i.e., Casimir and van der Waals attractions, are included in the theoretical model considering the correction due to the presence of liquid media between the interacting surfaces (three-layer model). To this end, a kind of microplate has been designed, i.e., a square microplate with all edges clamped supported. The strain gradient elasticity is employed to model the size-dependent structural behavior of the clamped microplate. To solve the nonlinear constitutive equation of the system, Extended Kantorovich Method, is employed and the pull-in parameter of the microplate are extracted. Impacts of the dispersion forces and size effect on the instability characteristics are discussed as well as the effect of ion concentration and potential ratio. It is found that the significant difference between the pull-in instability parameters in the modified strain gradient theory and the classical theory for thin microplates is merely due to the consideration of size effect parameter in the modified strain gradient theory. To confirm the validity of formulations, the numerical values of the results are compared. The results predicted via the aforementioned approach are in excellent agreement with those in the literature. Some new examples are solved to demonstrate the applicability of the procedure.

Fluorescein Diacetate Microplate Assay in Cell Viability Detection.

PubMed

Chen, Xi; Yang, Xiu-Ying; Fang, Lian-Hua; DU, Guan-Hua

2016-12-20

Objective To investigate the application of the fluorescein diacetate (FDA) microplate assay in cell viability detection. Methods Cells were seeded in a 96-well culture plate until detection. After incubated with FDA,the plate was detected by fluorescence microplate analyzer. The effects of FDA incubation duration,concentration,and other factors on the assay's accuracy and stability were assessed. We also compared the results of FDA with methyl thiazolyl(MTT) in terms of cell numbers and H 2 O 2 injury. Results Within 0-30 minutes,the fluorescence-cell number coefficient of FDA assay increased with duration and reached 0.99 in 27-30 minutes. The optimum concentration of final FDA in this study was 10-30 μg/ml. On cell viability detection,the result of FDA method was equivalent to MTT method. As to H 2 O 2 injury assay,the sensitivity of FDA method was superior to MTT on the higher concentration H 2 O 2 treatment due to a relative shorter duration for detection. Conclusion As a stable and reliable method,FDA is feasible for cell variability detection under varied conditions.

Measurement of Factor V Activity in Human Plasma Using a Microplate Coagulation Assay

PubMed Central

Tilley, Derek; Levit, Irina; Samis, John A.

2012-01-01

In response to injury, blood coagulation is activated and results in generation of the clotting protease, thrombin. Thrombin cleaves fibrinogen to fibrin which forms an insoluble clot that stops hemorrhage. Factor V (FV) in its activated form, FVa, is a critical cofactor for the protease FXa and accelerator of thrombin generation during fibrin clot formation as part of prothrombinase 1, 2. Manual FV assays have been described 3, 4, but they are time consuming and subjective. Automated FV assays have been reported 5-7, but the analyzer and reagents are expensive and generally provide only the clot time, not the rate and extent of fibrin formation. The microplate platform is preferred for measuring enzyme-catalyzed events because of convenience, time, cost, small volume, continuous monitoring, and high-throughput 8, 9. Microplate assays have been reported for clot lysis 10, platelet aggregation 11, and coagulation Factors 12, but not for FV activity in human plasma. The goal of the method was to develop a microplate assay that measures FV activity during fibrin formation in human plasma. This novel microplate method outlines a simple, inexpensive, and rapid assay of FV activity in human plasma. The assay utilizes a kinetic microplate reader to monitor the absorbance change at 405nm during fibrin formation in human plasma (Figure 1) 13. The assay accurately measures the time, initial rate, and extent of fibrin clot formation. It requires only μl quantities of plasma, is complete in 6 min, has high-throughput, is sensitive to 24-80pM FV, and measures the amount of unintentionally activated (1-stage activity) and thrombin-activated FV (2-stage activity) to obtain a complete assessment of its total functional activity (2-stage activity - 1-stage activity). Disseminated intravascular coagulation (DIC) is an acquired coagulopathy that most often develops from pre-existing infections 14. DIC is associated with a poor prognosis and increases mortality above the pre

Measurement of factor v activity in human plasma using a microplate coagulation assay.

PubMed

Tilley, Derek; Levit, Irina; Samis, John A

2012-09-09

In response to injury, blood coagulation is activated and results in generation of the clotting protease, thrombin. Thrombin cleaves fibrinogen to fibrin which forms an insoluble clot that stops hemorrhage. Factor V (FV) in its activated form, FVa, is a critical cofactor for the protease FXa and accelerator of thrombin generation during fibrin clot formation as part of prothrombinase (1, 2). Manual FV assays have been described (3, 4), but they are time consuming and subjective. Automated FV assays have been reported (5-7), but the analyzer and reagents are expensive and generally provide only the clot time, not the rate and extent of fibrin formation. The microplate platform is preferred for measuring enzyme-catalyzed events because of convenience, time, cost, small volume, continuous monitoring, and high-throughput (8, 9). Microplate assays have been reported for clot lysis (10), platelet aggregation (11), and coagulation Factors (12), but not for FV activity in human plasma. The goal of the method was to develop a microplate assay that measures FV activity during fibrin formation in human plasma. This novel microplate method outlines a simple, inexpensive, and rapid assay of FV activity in human plasma. The assay utilizes a kinetic microplate reader to monitor the absorbance change at 405 nm during fibrin formation in human plasma (Figure 1) (13). The assay accurately measures the time, initial rate, and extent of fibrin clot formation. It requires only μl quantities of plasma, is complete in 6 min, has high-throughput, is sensitive to 24-80 pM FV, and measures the amount of unintentionally activated (1-stage activity) and thrombin-activated FV (2-stage activity) to obtain a complete assessment of its total functional activity (2-stage activity - 1-stage activity). Disseminated intravascular coagulation (DIC) is an acquired coagulopathy that most often develops from pre-existing infections (14). DIC is associated with a poor prognosis and increases mortality

Nanochannels Photoelectrochemical Biosensor.

PubMed

Zhang, Nan; Ruan, Yi-Fan; Zhang, Li-Bin; Zhao, Wei-Wei; Xu, Jing-Juan; Chen, Hong-Yuan

2018-02-06

Nanochannels have brought new opportunities for biosensor development. Herein, we present the novel concept of a nanochannels photoelectrochemical (PEC) biosensor based on the integration of a unique Cu x O-nanopyramid-islands (NPIs) photocathode, an anodic aluminum oxide (AAO) membrane, and alkaline phosphatase (ALP) catalytic chemistry. The Cu x O-NPIs photocathode possesses good performance, and further assembly with AAO yields a designed architecture composed of vertically aligned, highly ordered nanoarrays on top of the Cu x O-NPIs film. After biocatalytic precipitation (BCP) was stimulated within the channels, the biosensor was used for the successful detection of ALP activity. This study has not only provided a novel paradigm for an unconventional nanochannels PEC biosensor, which can be used for general bioanalytical purposes, but also indicated that the new concept of nanochannel-semiconductor heterostructures is a step toward innovative biomedical applications.

Fast and Accurate Microplate Method (Biolog MT2) for Detection of Fusarium Fungicides Resistance/Sensitivity.

PubMed

Frąc, Magdalena; Gryta, Agata; Oszust, Karolina; Kotowicz, Natalia

2016-01-01

The need for finding fungicides against Fusarium is a key step in the chemical plant protection and using appropriate chemical agents. Existing, conventional methods of evaluation of Fusarium isolates resistance to fungicides are costly, time-consuming and potentially environmentally harmful due to usage of high amounts of potentially toxic chemicals. Therefore, the development of fast, accurate and effective detection methods for Fusarium resistance to fungicides is urgently required. MT2 microplates (Biolog(TM)) method is traditionally used for bacteria identification and the evaluation of their ability to utilize different carbon substrates. However, to the best of our knowledge, there is no reports concerning the use of this technical tool to determine fungicides resistance of the Fusarium isolates. For this reason, the objectives of this study are to develop a fast method for Fusarium resistance to fungicides detection and to validate the effectiveness approach between both traditional hole-plate and MT2 microplates assays. In presented study MT2 microplate-based assay was evaluated for potential use as an alternative resistance detection method. This was carried out using three commercially available fungicides, containing following active substances: triazoles (tebuconazole), benzimidazoles (carbendazim) and strobilurins (azoxystrobin), in six concentrations (0, 0.0005, 0.005, 0.05, 0.1, 0.2%), for nine selected Fusarium isolates. In this study, the particular concentrations of each fungicides was loaded into MT2 microplate wells. The wells were inoculated with the Fusarium mycelium suspended in PM4-IF inoculating fluid. Before inoculation the suspension was standardized for each isolates into 75% of transmittance. Traditional hole-plate method was used as a control assay. The fungicides concentrations in control method were the following: 0, 0.0005, 0.005, 0.05, 0.5, 1, 2, 5, 10, 25, and 50%. Strong relationships between MT2 microplate and traditional hole

Fast and Accurate Microplate Method (Biolog MT2) for Detection of Fusarium Fungicides Resistance/Sensitivity

PubMed Central

Frąc, Magdalena; Gryta, Agata; Oszust, Karolina; Kotowicz, Natalia

2016-01-01

The need for finding fungicides against Fusarium is a key step in the chemical plant protection and using appropriate chemical agents. Existing, conventional methods of evaluation of Fusarium isolates resistance to fungicides are costly, time-consuming and potentially environmentally harmful due to usage of high amounts of potentially toxic chemicals. Therefore, the development of fast, accurate and effective detection methods for Fusarium resistance to fungicides is urgently required. MT2 microplates (BiologTM) method is traditionally used for bacteria identification and the evaluation of their ability to utilize different carbon substrates. However, to the best of our knowledge, there is no reports concerning the use of this technical tool to determine fungicides resistance of the Fusarium isolates. For this reason, the objectives of this study are to develop a fast method for Fusarium resistance to fungicides detection and to validate the effectiveness approach between both traditional hole-plate and MT2 microplates assays. In presented study MT2 microplate-based assay was evaluated for potential use as an alternative resistance detection method. This was carried out using three commercially available fungicides, containing following active substances: triazoles (tebuconazole), benzimidazoles (carbendazim) and strobilurins (azoxystrobin), in six concentrations (0, 0.0005, 0.005, 0.05, 0.1, 0.2%), for nine selected Fusarium isolates. In this study, the particular concentrations of each fungicides was loaded into MT2 microplate wells. The wells were inoculated with the Fusarium mycelium suspended in PM4-IF inoculating fluid. Before inoculation the suspension was standardized for each isolates into 75% of transmittance. Traditional hole-plate method was used as a control assay. The fungicides concentrations in control method were the following: 0, 0.0005, 0.005, 0.05, 0.5, 1, 2, 5, 10, 25, and 50%. Strong relationships between MT2 microplate and traditional hole

[Change in soil enzymes activities after adding biochar or straw by fluorescent microplate method].

PubMed

Zhang, Yu-Lan; Chen, Li-Jun; Duan, Zheng-Hu; Wu, Zhi-Jie; Sun, Cai-Xia; Wang, Jun-Yu

2014-02-01

The present work was aimed to study soil a-glucosidase and beta-glucosidase activities of and red soils based on fluorescence detection method combined with 96 microplates with TECAN Infinite 200 Multi-Mode Microplate Reader. We added biochar or straw (2.5 g air dry sample/50g air dry soil sample) into and red soils and the test was carried under fixed temperature and humidity condition (25 degrees C, 20% soil moisture content). The results showed that straw addition enhances soil alpha-glucosidase and beta-glucosidase activities, beta-glucosidase activity stimulated by rice straw treatment was higher than that of corn straw treatment, and activity still maintains strong after 40 days, accounting for increasing soil carbon transformation with straw inputting. Straw inputting increased soil nutrients contents and may promote microbial activity, which also lead to the increase oin enzyme Straw inputting increased soil nutrients contents and may promote microbial activity, which also lead to the increase oin enzyme activities. Different effects of straw kinds may be related to material source that needs further research. However, biochar inputting has little effect on soil alpha-glucosidase and beta-glucosidase activity. Biochar contains less available nutrients than straw and have degradation-resistant characteristics. Compared with the conventional spectrophotometric method, fluorescence microplate method is more sensitive to soil enzyme activities in suspension liquid, which can be used in a large number of samples. In brief, fluorescence microplate method is fast, accurate, and simple to determine soil enzymes activities.

Large-scale preparation of shape controlled SnO and improved capacitance for supercapacitors: from nanoclusters to square microplates.

PubMed

Wang, Lu; Ji, Hongmei; Zhu, Feng; Chen, Zhi; Yang, Yang; Jiang, Xuefan; Pinto, João; Yang, Gang

2013-08-21

Here, we first provide a facile ultrasonic-assisted synthesis of SnO using SnCl2 and the organic solvent of ethanolamine (ETA). The moderate alkalinity of ETA and ultrasound play very important roles in the synthesis of SnO. After the hydrolysis of the intermediate of ETA-Sn(II), the as-synthesized SnO nanoclusters undergo assembly, amalgamation, and preferential growth to microplates in hydrothermal treatment. The as-synthesized SnO was characterized by scanning electron microscopy (SEM), transmission electron microscopy (TEM), high-resolution transmission electron microscopy (HRTEM), ultraviolet-visible absorption spectroscopy (UV-vis) and X-ray diffraction (XRD). To explore its potential applications in energy storage, SnO was fabricated into a supercapacitor electrode and characterized by cyclic voltammetry (CV), electrochemical impedance spectroscopy (EIS), and galvanostatic charge-discharge measurements. The as-synthesized SnO exhibits remarkable pseudocapacitive activity including high specific capacitance (208.9 F g(-1) at 0.1 A g(-1)), good rate capability (65.8 F g(-1) at 40 A g(-1)), and excellent cycling stability (retention 119.3% after 10,000 cycles) for application in supercapacitors. The capacitive behavior of SnO with various crystal morphologies was observed by fitted EIS using an equivalent circuit. The novel synthetic route for SnO is a convenient and potential way to large-scale production of microplates which is expected to be applicable in the synthesis of other metal oxide nanoparticles.

A simple technique to reduce evaporation of crystallization droplets by using plate lids with apertures for adding liquids

DOE Office of Scientific and Technical Information (OSTI.GOV)

Zipper, Lauren E.; Binghamton University, 4400 Vestal Parkway East, Vestal, NY 13902; Aristide, Xavier

This article describes the use of evaporation control lids that are fitted to crystallization plates to improve the reproducibility of trials using as little as 5 nl. The plate lids contain apertures which are large enough for the transfer of protein containing droplets, but small enough to greatly reduce the rate of evaporation during the time needed to prepare the plate. A method is described for using plate lids to reduce evaporation in low-volume vapor-diffusion crystallization experiments. The plate lids contain apertures through which the protein and precipitants were added to different crystallization microplates (the reservoir was filled before fittingmore » the lids). Plate lids were designed for each of these commonly used crystallization microplates. This system minimizes the dehydration of crystallization droplets containing just a few nanolitres of protein and precipitant, and results in more reproducible diffraction from the crystals. For each lid design, changes in the weight of the plates were used to deduce the rate of evaporation under different conditions of temperature, air movement, droplet size and precipitant. For comparison, the state of dehydration was also visually assessed throughout the experiment. Finally, X-ray diffraction methods were used to compare the diffraction of protein crystals that were conventionally prepared against those that were prepared on plates with plate lids. The measurements revealed that the plate lids reduced the rate of evaporation by 63–82%. Crystals grown in 5 nl drops that were set up with plate lids diffracted to higher resolution than similar crystals from drops that were set up without plate lids. The results demonstrate that plate lids can be instrumental for improving few-nanolitre crystallizations.« less

[Microplate luminometry for toxicity bioassay of chemicals on luciferase].

PubMed

Ge, Hui-Lin; Liu, Shu-Shen; Chen, Fu; Luo, Jin-Hui; Lü, Dai-Zhu; Su, Bing-Xia

2013-10-01

A new microplate luminometry for the toxicity bioassay of chemicals on firefly luciferase, was developed using the multifunctional microplate reader (SpectraMax M5) to measure the luminous intensity of luciferase. Efects of luciferase concentration, luciferin concentration, ATP concentration, pH, temperature, and reaction time on the luminescence were systematically investigated. It was found that ATP exerted a biphasic response on the luciferase luminescence and the maximum relative light units (RLU) occurred at an ATP concentration of 1.1 x 10(-4) mol x L(-1). The method was successfully employed in the toxic effect test of NaF, NaCl, KBr and NaBF4 on luciferase. Using nonlinear least square technique, the dose-response curves (DRC) of the 4 chemicals were accurately fitted with the coefficient of determination (R2) between the fitted and observed responses being greater than 0.99. The median effective concentration (EC50) of the 4 chemicals were accurately measured from the DRC models. Compared with some literatures, the bioassay is a fast easy-operate and cost-effective method with high accuracy.

Coupling the Torpedo microplate-receptor binding assay with mass spectrometry to detect cyclic imine neurotoxins.

PubMed

Aráoz, Rómulo; Ramos, Suzanne; Pelissier, Franck; Guérineau, Vincent; Benoit, Evelyne; Vilariño, Natalia; Botana, Luis M; Zakarian, Armen; Molgó, Jordi

2012-12-04

Cyclic imine neurotoxins constitute an emergent family of neurotoxins of dinoflagellate origin that are potent antagonists of nicotinic acetylcholine receptors. We developed a target-directed functional method based on the mechanism of action of competitive agonists/antagonists of nicotinic acetylcholine receptors for the detection of marine cyclic imine neurotoxins. The key step for method development was the immobilization of Torpedo electrocyte membranes rich in nicotinic acetylcholine receptors on the surface of microplate wells and the use of biotinylated-α-bungarotoxin as tracer. Cyclic imine neurotoxins competitively inhibit biotinylated-α-bungarotoxin binding to Torpedo-nicotinic acetylcholine receptors in a concentration-dependent manner. The microplate-receptor binding assay allowed rapid detection of nanomolar concentrations of cyclic imine neurotoxins directly in shellfish samples. Although highly sensitive and specific for the detection of neurotoxins targeting nicotinic acetylcholine receptors as a class, the receptor binding assay cannot identify a given analyte. To address the low selectivity of the microplate-receptor binding assay, the cyclic imine neurotoxins tightly bound to the coated Torpedo nicotinic receptor were eluted with methanol, and the chemical nature of the eluted ligands was identified by mass spectrometry. The immobilization of Torpedo electrocyte membranes on the surface of microplate wells proved to be a high-throughput format for the survey of neurotoxins targeting nicotinic acetylcholine receptors directly in shellfish matrixes with high sensitivity and reproducibility.

Coupling the Torpedo Microplate-Receptor Binding Assay with Mass Spectrometry to Detect Cyclic Imine Neurotoxins

PubMed Central

Aráoz, Rómulo; Ramos, Suzanne; Pelissier, Franck; Guérineau, Vincent; Benoit, Evelyne; Vilariño, Natalia; Botana, Luis M.; Zakarian, Armen; Molgó, Jordi

2014-01-01

Cyclic imine neurotoxins constitute an emergent family of neurotoxins of dinoflagellate origin that are potent antagonists of nicotinic acetylcholine receptors. We developed a target-directed functional method based on the mechanism of action of competitive agonists/antagonists of nicotinic acetylcholine receptors for the detection of marine cyclic imine neurotoxins. The key step for method development was the immobilization of Torpedo electrocyte membranes rich in nicotinic acetylcholine receptors on the surface of microplate wells and the use of biotinylated-α-bungarotoxin as tracer. Cyclic imine neurotoxins competitively inhibit biotinylated-α-bungarotoxin binding to Torpedo-nicotinic acetylcholine receptors in a concentration-dependent manner. The microplate-receptor binding assay allowed rapid detection of nanomolar concentrations of cyclic imine neurotoxins directly in shellfish samples. Although highly sensitive and specific for the detection of neurotoxins targeting nicotinic acetylcholine receptors as a class, the receptor binding assay cannot identify a given analyte. To address the low selectivity of the microplate-receptor binding assay, the cyclic imine neurotoxins tightly bound to the coated Torpedo nicotinic receptor were eluted with methanol, and the chemical nature of the eluted ligands was identified by mass spectrometry. The immobilization of Torpedo electrocyte membranes on the surface of microplate wells proved to be a high-throughput format for the survey of neurotoxins targeting nicotinic acetylcholine receptors directly in shellfish matrixes with high sensitivity and reproducibility. PMID:23131021

Plasmonic Biosensors

PubMed Central

Hill, Ryan T.

2015-01-01

The unique optical properties of plasmon resonant nanostructures enable exploration of nanoscale environments using relatively simple optical characterization techniques. For this reason, the field of plasmonics continues to garner the attention of the biosensing community. Biosensors based on propagating surface plasmon resonances (SPRs) in films are the most well-recognized plasmonic biosensors, but there is great potential for the new, developing technologies to surpass the robustness and popularity of film-based SPR sensing. This review surveys the current plasmonic biosensor landscape with emphasis on the basic operating principles of each plasmonic sensing technique and the practical considerations when developing a sensing platform with the various techniques. The “gold standard” film SPR technique is reviewed briefly, but special emphasis is devoted to the up-and-coming LSPR-based and plasmonically coupled sensor technology. PMID:25377594

Dependence of seed layer thickness on sensitivity of nano-ZnO cholesterol biosensor

NASA Astrophysics Data System (ADS)

Lu, Yang-Ming; Wang, Po-Chin; Tang, Jian-Fu; Chu, Sheng-Yuan

2017-01-01

The anemone-like ZnO nanostructures have been synthesized by hydrothermal method and were further adsorbed immobilized cholesterol oxidase (ChOx) as a nano-biosensor. In this study, the sensitivity of biosensor were improved by varying the thickness of the ZnO seed layer. The SEM analysis showed changes in thickness of seed layer will not affect the morphologies of anemone-like ZnO nanostructures. The X-ray Diffraction patterns showed that the (002) plane of anemone-like ZnO grown on various thickness of the seed layer was more prouded than other crystal plane. Abioelectrode (ChOx/ZnO/ITO/glass) grown on the 30nm of ZnO seed layer with high sensitivity of 57.533μAmM-1cm-2 (1.488 μA (mg/dl) -1cm-2), a wide sensitive range from 25 to 500 mg/dl. It is concluded that the thinner sputtered ZnO seed layer for growing anemone-like ZnO nanostructure can effectively improve the sensitivity of the ZnO biosensor.

A survey of the 2001 to 2005 quartz crystal microbalance biosensor literature: applications of acoustic physics to the analysis of biomolecular interactions.

PubMed

Cooper, Matthew A; Singleton, Victoria T

2007-01-01

The widespread exploitation of biosensors in the analysis of molecular recognition has its origins in the mid-1990s following the release of commercial systems based on surface plasmon resonance (SPR). More recently, platforms based on piezoelectric acoustic sensors (principally 'bulk acoustic wave' (BAW), 'thickness shear mode' (TSM) sensors or 'quartz crystal microbalances' (QCM)), have been released that are driving the publication of a large number of papers analysing binding specificities, affinities, kinetics and conformational changes associated with a molecular recognition event. This article highlights salient theoretical and practical aspects of the technologies that underpin acoustic analysis, then reviews exemplary papers in key application areas involving small molecular weight ligands, carbohydrates, proteins, nucleic acids, viruses, bacteria, cells and lipidic and polymeric interfaces. Key differentiators between optical and acoustic sensing modalities are also reviewed. Copyright (c) 2007 John Wiley & Sons, Ltd.

Capacitive Biosensors and Molecularly Imprinted Electrodes.

PubMed

Ertürk, Gizem; Mattiasson, Bo

2017-02-17

Capacitive biosensors belong to the group of affinity biosensors that operate by registering direct binding between the sensor surface and the target molecule. This type of biosensors measures the changes in dielectric properties and/or thickness of the dielectric layer at the electrolyte/electrode interface. Capacitive biosensors have so far been successfully used for detection of proteins, nucleotides, heavy metals, saccharides, small organic molecules and microbial cells. In recent years, the microcontact imprinting method has been used to create very sensitive and selective biorecognition cavities on surfaces of capacitive electrodes. This chapter summarizes the principle and different applications of capacitive biosensors with an emphasis on microcontact imprinting method with its recent capacitive biosensor applications.

Surface derivatization with spacer molecules on glutaraldehyde-activated amino-microplates for covalent immobilization of β-glucosidase

NASA Astrophysics Data System (ADS)

Zhang, Yaodong; Zhang, Yun; Jiang, Juanjuan; Li, Li; Yu, Caihong; Hei, Tingting

2011-01-01

Protein molecules immobilized on a hydrophobic polystyrene microplate by passive adsorption lose their activity and suffer considerable denaturation. In this paper, we report a thorough evaluation of a protocol for enzyme immobilization on a microplate with relatively inexpensive reagents, involving glutaraldehyde coupling and spacer molecules, and employing β-glucosidase as a model enzyme. The recommended conditions for the developed method include 2.5% glutaraldehyde to activate the reaction, 1% chitosan in an HAc solution to increase the binding capacity, 2% bovine serum albumin to block non-specific binding sites, and 0.1 M NaBH4 to stabilize Schiff's base intermediates. Using this method, the amount of β-glucosidase immobilized on amino-microplate was 24-fold with chitosan than without spacer molecules. The procedure is efficient and quite simple, and may thus have potential applications in biosensing and bioreactor systems.

Improved high-throughput quantification of luminescent microplate assays using a common Western-blot imaging system.

PubMed

Hawkins, Liam J; Storey, Kenneth B

2017-01-01

Common Western-blot imaging systems have previously been adapted to measure signals from luminescent microplate assays. This can be a cost saving measure as Western-blot imaging systems are common laboratory equipment and could substitute a dedicated luminometer if one is not otherwise available. One previously unrecognized limitation is that the signals captured by the cameras in these systems are not equal for all wells. Signals are dependent on the angle of incidence to the camera, and thus the location of the well on the microplate. Here we show that: •The position of a well on a microplate significantly affects the signal captured by a common Western-blot imaging system from a luminescent assay.•The effect of well position can easily be corrected for.•This method can be applied to commercially available luminescent assays, allowing for high-throughput quantification of a wide range of biological processes and biochemical reactions.

Cellphone-based hand-held microplate reader for point-of-care ELISA testing (Conference Presentation)

NASA Astrophysics Data System (ADS)

Berg, Brandon; Cortazar, Bingen; Tseng, Derek; Ozkan, Haydar; Feng, Steve; Wei, Qingshan; Chan, Raymond Y.; Burbano, Jordi; Farooqui, Qamar; Lewinski, Michael; Di Carlo, Dino; Garner, Omai B.; Ozcan, Aydogan

2016-03-01

Enzyme-linked immunosorbent assay (ELISA) in a microplate format has been a gold standard first-line clinical test for diagnosis of various diseases including infectious diseases. However, this technology requires a relatively large and expensive multi-well scanning spectrophotometer to read and quantify the signal from each well, hindering its implementation in resource-limited-settings. Here, we demonstrate a cost-effective and handheld smartphone-based colorimetric microplate reader for rapid digitization and quantification of immunoserology-related ELISA tests in a conventional 96-well plate format at the point of care (POC). This device consists of a bundle of 96 optical fibers to collect the transmitted light from each well of the microplate and direct all the transmission signals from the wells onto the camera of the mobile-phone. Captured images are then transmitted to a remote server through a custom-designed app, and both quantitative and qualitative diagnostic results are returned back to the user within ~1 minute per 96-well plate by using a machine learning algorithm. We tested this mobile-phone based micro-plate reader in a clinical microbiology lab using FDA-approved mumps IgG, measles IgG, and herpes simplex virus IgG (HSV-1 and HSV-2) ELISA tests on 1138 remnant patient samples (roughly 50% training and 50% testing), and achieved an overall accuracy of ~99% or higher for each ELISA test. This handheld and cost-effective platform could be immediately useful for large-scale vaccination monitoring in low-infrastructure settings, and also for other high-throughput disease screening applications at POC.

Plasmonic biosensors.

PubMed

Hill, Ryan T

2015-01-01

The unique optical properties of plasmon resonant nanostructures enable exploration of nanoscale environments using relatively simple optical characterization techniques. For this reason, the field of plasmonics continues to garner the attention of the biosensing community. Biosensors based on propagating surface plasmon resonances (SPRs) in films are the most well-recognized plasmonic biosensors, but there is great potential for the new, developing technologies to surpass the robustness and popularity of film-based SPR sensing. This review surveys the current plasmonic biosensor landscape with emphasis on the basic operating principles of each plasmonic sensing technique and the practical considerations when developing a sensing platform with the various techniques. The 'gold standard' film SPR technique is reviewed briefly, but special emphasis is devoted to the up-and-coming localized surface plasmon resonance and plasmonically coupled sensor technology. © 2014 Wiley Periodicals, Inc.

Introduction to biosensors

PubMed Central

Bhalla, Nikhil; Jolly, Pawan; Formisano, Nello

2016-01-01

Biosensors are nowadays ubiquitous in biomedical diagnosis as well as a wide range of other areas such as point-of-care monitoring of treatment and disease progression, environmental monitoring, food control, drug discovery, forensics and biomedical research. A wide range of techniques can be used for the development of biosensors. Their coupling with high-affinity biomolecules allows the sensitive and selective detection of a range of analytes. We give a general introduction to biosensors and biosensing technologies, including a brief historical overview, introducing key developments in the field and illustrating the breadth of biomolecular sensing strategies and the expansion of nanotechnological approaches that are now available. PMID:27365030

Seismicity of the Adriatic microplate

USGS Publications Warehouse

Console, R.; Di, Giovambattista R.; Favali, P.; Presgrave, B.W.; Smriglio, G.

1993-01-01

The Adriatic microplate was previously considered to be a unique block, tectonically active only along its margins. The seismic sequences that took place in the basin from 1986 to 1990 give new information about the geodynamics of this area. Three subsets of well recorded events were relocated by the joint hypocentre determination technique. On the whole, this seismic activity was concentrated in a belt crossing the southern Adriatic sea around latitude 42??, in connection with regional E-W fault systems. Some features of this seismicity, similar to those observed in other well known active margins of the Adriatic plate, support a model of a southern Adriatic lithospheric block, detached from the Northern one. Other geophysical information provides evidence of a transitional zone at the same latitude. ?? 1993.

Microplate Bioassay for Determining Substrate Selectivity of "Candida rugosa" Lipase

ERIC Educational Resources Information Center

Wang, Shi-zhen; Fang, Bai-shan

2012-01-01

Substrate selectivity of "Candida rugosa" lipase was tested using "p"-nitrophenyl esters of increasing chain length (C[subscript 1], C[subscript 7], C[subscript 15]) using the high-throughput screening method. A fast and easy 96-well microplate bioassay was developed to help students learn and practice biotechnological specificity screen. The…

Graphene-based biosensors.

PubMed

Szunerits, Sabine; Boukherroub, Rabah

2018-06-06

Reliable data obtained from analysis of DNA, proteins, bacteria and other disease-related molecules or organisms in biological samples have become a fundamental and crucial part of human health diagnostics and therapy. The development of non-invasive tests that are rapid, sensitive, specific and simple would allow patient discomfort to be prevented, delays in diagnosis to be avoided and the status of a disease to be followed up. Bioanalysis is thus a progressive discipline for which the future holds many exciting opportunities. The use of biosensors for the early diagnosis of diseases has become widely accepted as a point-of-care diagnosis with appropriate specificity in a short time. To allow a reliable diagnosis of a disease at an early stage, highly sensitive biosensors are required as the corresponding biomarkers are generally expressed at very low concentrations. In the past 50 years, various biosensors have been researched and developed encompassing a wide range of applications. This contrasts the limited number of commercially available biosensors. When it comes to sensing of biomarkers with the required picomolar (pM) sensitivity for real-time sensing of biological samples, only a handful of sensing systems have been proposed, and these are often rather complex and costly. Lately, graphene-based materials have been considered as superior over other nanomaterials for the development of sensitive biosensors. The advantages of graphene-based sensor interfaces are numerous, including enhanced surface loading of the desired ligand due to the high surface-to-volume ratio, excellent conductivity and a small band gap that is beneficial for sensitive electrical and electrochemical read-outs, as well as tunable optical properties for optical read-outs such as fluorescence and plasmonics. In this paper, we review the advances made in recent years on graphene-based biosensors in the field of medical diagnosis.

Graphene-based field-effect transistor biosensors

DOEpatents

Chen; , Junhong; Mao, Shun; Lu, Ganhua

2017-06-14

The disclosure provides a field-effect transistor (FET)-based biosensor and uses thereof. In particular, to FET-based biosensors using thermally reduced graphene-based sheets as a conducting channel decorated with nanoparticle-biomolecule conjugates. The present disclosure also relates to FET-based biosensors using metal nitride/graphene hybrid sheets. The disclosure provides a method for detecting a target biomolecule in a sample using the FET-based biosensor described herein.

Ejection of small droplet from microplate using focused ultrasound

NASA Astrophysics Data System (ADS)

Tanaka, Hiroki; Mizuno, Yosuke; Nakamura, Kentaro

2017-08-01

We discussed an ultrasonic system for single-droplet ejection from a microplate, which is one of the basic and important procedures in the noncontact handling of droplets in air. In this system, a 1.5 MHz concave transducer located below the microplate is used for chasing the liquid surface through a pulse echo method, and also for the ejection of a 1 µL single droplet by the burst of focused ultrasound. We investigated the relationship between the droplet ejection characteristics, the distance from the transducer to the surface of liquid, the material property, and the excitation condition of the focused ultrasonic transducer. It was verified that the optimal position of the transducer was off the focal point of sound pressure by ±1 mm, because the sound intensity had to be controlled to eject a single droplet. Subsequently, we confirmed experimentally that the ejected droplet volume linearly depended on the surface tension of the liquid, and that the droplet volume and ejection velocity were determined by the Webber number, Reynolds number, and Ohnesolge number. In addition, by optimizing the duration of the burst ultrasound, the droplet volume and ejection velocity were controlled.

A microplate assay for DNA damage determination (fast micromethod).

PubMed

Batel, R; Jaksić, Z; Bihari, N; Hamer, B; Fafandel, M; Chauvin, C; Schröder, H C; Müller, W E; Zahn, R K

1999-06-01

A rapid and convenient procedure for DNA damage determination in cell suspensions and solid tissues on single microplates was developed. The procedure is based on the ability of commercially available fluorochromes to interact preferentially with dsDNA in the presence of ssDNA, RNA, and proteins at high pH (>12.0), thus allowing direct measurements of DNA denaturation without sample handling or stepwise DNA separations. The method includes a simple and rapid 40-min sample lysis in the presence of EDTA, SDS, and high urea concentration at pH 10, followed by time-dependent DNA denaturation at pH 12.4 after NaOH addition. The time course and the extent of DNA denaturation is followed in a microplate fluorescence reader at room temperature for less than 1 h. The method requires only 30 ng DNA per single well and could conveniently be used whenever fast analysis of DNA integrity in small samples has to be done, e.g., in patients' lymphocytes after irradiation or chemotherapy (about 3000 cells per sample), in solid tissues or biopsies after homogenization (about 25 microg tissue per well), or in environmental samples for genotoxicity assessment. Copyright 1999 Academic Press.

Improving the binding efficiency of quartz crystal microbalance biosensors by applying the electrothermal effect

PubMed Central

Huang, Yao-Hung; Chang, Jeng-Shian; Chao, Sheng D.; Wu, Kuang-Chong; Huang, Long-Sun

2014-01-01

A quartz crystal microbalance (QCM) serving as a biosensor to detect the target biomolecules (analytes) often suffers from the time consuming process, especially in the case of diffusion-limited reaction. In this experimental work, we modify the reaction chamber of a conventional QCM by integrating into the multi-microelectrodes to produce electrothermal vortex flow which can efficiently drive the analytes moving toward the sensor surface, where the analytes were captured by the immobilized ligands. The microelectrodes are placed on the top surface of the chamber opposite to the sensor, which is located on the bottom of the chamber. Besides, the height of reaction chamber is reduced to assure that the suspended analytes in the fluid can be effectively drived to the sensor surface by induced electrothermal vortex flow, and also the sample costs are saved. A series of frequency shift measurements associated with the adding mass due to the specific binding of the analytes in the fluid flow and the immobilized ligands on the QCM sensor surface are performed with or without applying electrothermal effect (ETE). The experimental results show that electrothermal vortex flow does effectively accelerate the specific binding and make the frequency shift measurement more sensible. In addition, the images of the binding surfaces of the sensors with or without applying electrothermal effect are taken through the scanning electron microscopy. By comparing the images, it also clearly indicates that ETE does raise the specific binding of the analytes and ligands and efficiently improves the performance of the QCM sensor. PMID:25538808

Characterization of Textile-Insulated Capacitive Biosensors

PubMed Central

Ng, Charn Loong; Reaz, Mamun Bin Ibne

2017-01-01

Capacitive biosensors are an emerging technology revolutionizing wearable sensing systems and personal healthcare devices. They are capable of continuously measuring bioelectrical signals from the human body while utilizing textiles as an insulator. Different textile types have their own unique properties that alter skin-electrode capacitance and the performance of capacitive biosensors. This paper aims to identify the best textile insulator to be used with capacitive biosensors by analysing the characteristics of 6 types of common textile materials (cotton, linen, rayon, nylon, polyester, and PVC-textile) while evaluating their impact on the performance of a capacitive biosensor. A textile-insulated capacitive (TEX-C) biosensor was developed and validated on 3 subjects. Experimental results revealed that higher skin-electrode capacitance of a TEX-C biosensor yields a lower noise floor and better signal quality. Natural fabric such as cotton and linen were the two best insulating materials to integrate with a capacitive biosensor. They yielded the lowest noise floor of 2 mV and achieved consistent electromyography (EMG) signals measurements throughout the performance test. PMID:28287493

Molecule counting with alkanethiol and DNA immobilized on gold microplates for extended gate FET.

PubMed

Cao, Zhong; Xiao, Zhong-Liang; Zhang, Ling; Luo, Dong-Mei; Kamahori, Masao; Shimoda, Maki

2013-04-01

Several molecule counting methods based on electrochemical characterization of alkanethiol and thiolated single-stranded oligonucleotide (HS-ssDNA) immobilized on gold microplates, which were used as extended gates of field effect transistors (FETs), have been investigated in this paper. The surface density of alkanethiol and DNA monolayers on gold microplates were quantitatively evaluated from the reductive desorption charge by using cyclic voltammetry (CV) and fast CV (FCV) methods in strong alkali solution. Typically, the surface density of 6-hydroxy-1-hexanethiol (6-HHT) was evaluated to be 4.639 molecules/nm(2), and the 28 base-pair dsDNA about 1.226-4.849 molecules/100 nm(2) on Au microplates after post-treatment with 6-HHT. The behaviors on surface potential and capacitance of different aminoalkanethiols on Au microplates were measured in 0.1 mol/L Na2SO4 and 10 mmol/L Tris-HCl (pH=7.4) solutions, indicating that the surface potential increases and the double-layer capacitance decreases with the length of carbon chain increased for the thiol monolayers, which obey a physics relationship for a capacitor. Comparably, a simple sensing method based on the electronic signals of biochemical reaction events on DNA immobilization and hybridization at the Au surface of the extended gate FET (EGFET) was developed, with which the surface density of the hybridized dsDNA on the gold surface of the EGFET was evaluated to be 1.36 molecules per 100 nm(2), showing that the EGFET is a promising sensing biochip for DNA molecule counting. Copyright © 2012 Elsevier B.V. All rights reserved.

ZnO-Based Amperometric Enzyme Biosensors

PubMed Central

Zhao, Zhiwei; Lei, Wei; Zhang, Xiaobing; Wang, Baoping; Jiang, Helong

2010-01-01

Nanostructured ZnO with its unique properties could provide a suitable microenvironment for immobilization of enzymes while retaining their biological activity, and thus lead to an expanded use of this nanomaterial for the construction of electrochemical biosensors with enhanced analytical performance. ZnO-based enzyme electrochemical biosensors are summarized in several tables for an easy overview according to the target biosensing analyte (glucose, hydrogen peroxide, phenol and cholesterol), respectively. Moreover, recent developments in enzyme electrochemical biosensors based on ZnO nanomaterials are reviewed with an emphasis on the fabrications and features of ZnO, approaches for biosensor construction (e.g., modified electrodes and enzyme immobilization) and biosensor performances. PMID:22205864

Intra-laboratory validation of microplate methods for total phenolic content and antioxidant activity on polyphenolic extracts, and comparison with conventional spectrophotometric methods.

PubMed

Bobo-García, Gloria; Davidov-Pardo, Gabriel; Arroqui, Cristina; Vírseda, Paloma; Marín-Arroyo, María R; Navarro, Montserrat

2015-01-01

Total phenolic content (TPC) and antioxidant activity (AA) assays in microplates save resources and time, therefore they can be useful to overcome the fact that the conventional methods are time-consuming, labour intensive and use large amounts of reagents. An intra-laboratory validation of the Folin-Ciocalteu microplate method to measure TPC and the 2,2-diphenyl-1-picrylhydrazyl (DPPH) microplate method to measure AA was performed and compared with conventional spectrophotometric methods. To compare the TPC methods, the confidence intervals of a linear regression were used. In the range of 10-70 mg L(-1) of gallic acid equivalents (GAE), both methods were equivalent. To compare the AA methodologies, the F-test and t-test were used in a range from 220 to 320 µmol L(-1) of Trolox equivalents. Both methods had homogeneous variances, and the means were not significantively different. The limits of detection and quantification for the TPC microplate method were 0.74 and 2.24 mg L(-1) GAE and for the DPPH 12.07 and 36.58 µmol L(-1) of Trolox equivalents. The relative standard deviation of the repeatability and reproducibility for both microplate methods were ≤ 6.1%. The accuracy ranged from 88% to 100%. The microplate and the conventional methods are equals in a 95% confidence level. © 2014 Society of Chemical Industry.

Fiber optic choline biosensor

NASA Astrophysics Data System (ADS)

Wang, Hong; Cao, Xiaojian; Jia, Ke; Chai, Xueting; Lu, Hua; Lu, Zuhong

2001-10-01

A fiber optic fluorescence biosensor for choline is introduced in this paper. Choline is an important neurotransmitter in mammals. Due to the growing needs for on-site clinical monitoring of the choline, much effect has been devoted to develop choline biosensors. Fiber-optic fluorescence biosensors have many advantages, including miniaturization, flexibility, and lack of electrical contact and interference. The choline fiber-optic biosensor we designed implemented a bifurcated fiber to perform fluorescence measurements. The light of the blue LED is coupled into one end of the fiber as excitation and the emission spectrum from sensing film is monitored by fiber-spectrometer (S2000, Ocean Optics) through the other end of the fiber. The sensing end of the fiber is coated with Nafion film dispersed with choline oxidase and oxygen sensitive luminescent Ru(II) complex (Tris(2,2'-bipyridyl)dichlororuthenium(II), hexahydrate). Choline oxidase catalyzes the oxidation of choline to betaine and hydrogen peroxide while consuming oxygen. The fluorescence intensity of oxygen- sensitive Ru(II) are related to the choline concentration. The response of the fiber-optic sensor in choline solution is represented and discussed. The result indicates a low-cost, high-performance, portable choline biosensor.

A Cenozoic tectonic model for Southeast Asia - microplates and basins

DOE Office of Scientific and Technical Information (OSTI.GOV)

Maher, K.A.

1995-04-01

A computer-assisted Cenozoic tectonic model was built for Southeast Asia and used to construct 23 base maps, 2 to 6 million years apart. This close temporal spacing was necessary to constrain all the local geometric shifts in a consistent and geologically feasible fashion. More than a hundred individual blocks were required to adequately treat Cenozoic microplate processes at a basic level. The reconstructions show tectonic evolution to be characterized by long periods of gradual evolution, interrupted by brief, widespread episodes of reorganization in fundamental plate geometries and kinematics. These episodes are triggered by major collisions, or by accumulation of smallermore » changes. The model takes into account difficulties inherent in the region. The Pacific and Indo-Australian plates and their predecessors have driven westward and northward since the late Paleozoic, towards each other and the relatively stationary backstop of Asia. Southeast Asia is therefore the result of a long-lived, complex process of convergent tectonics, making it difficult to reconstruct tectonic evolution as much of the continental margin and sea floor spreading record was erased. In addition, the region has been dominated by small-scale microplate processes with short time scales and internal deformation, taking place in rapidly evolving and more ductile buffer zones between the major rigid plate systems. These plate interaction zones have taken up much of the relative motion between the major plates. Relatively ephemeral crustal blocks appear and die within the buffer zones, or accrete to and disperse from the margins of the major plate systems. However, such microplate evolution is the dominant factor in Cenozoic basin evolution. This detailed testonic model aids in comprehension and prediction of basin development, regional hydrocarbon habitat, and petroleum systems.« less

Microplate-based method to screen inhibitors of isozymes of cyclic nucleotide phosphodiesterase fused to SUMO.

PubMed

Chen, Chunyan; Liu, Miaomiao; Wu, Jing; Yang, Xiaolan; Hu, Xiaolei; Pu, Jun; Long, Gaobo; Xie, Yanling; Jiang, Hairong; Yuan, Yonghua; Liao, Fei

2014-12-01

The feasibility for microplate-based screening of inhibitors of isozymes of cyclic nucleotide phosphodiesterase (PDE) was tested via the coupled action of a phosphatase on adenosine-5'-monophosphate and an improved malachite green assay of phosphate. Human full-length PDE4B2 and truncated mutant (152-528aa) were expressed in Escherichia coli via fusion to SUMO, which after purification through Ni-NTA column exhibited specific activities >0.017 U mg(-1). In the presence of proteins <30 mg L(-1), absorbance for 10 µΜ phosphate was measurable; a PDE isozyme of specific activity over 0.008 U mg(-1) after reaction for 20 min thus suited for microplate-based screening of inhibitors. By using Biotek ELX 800 microplate reader, affinities of two forms of PEDE4B2 for cAMP, rolipram and papaverine varied over three magnitudes and were consistent with those by routine assay, respectively. Hence, the proposed method was promising for high-throughput-screening of inhibitors of phosphate-releasing enzymes bearing specific activities over 0.008 U mg(-1).

Label-Free Biosensor Imaging on Photonic Crystal Surfaces.

PubMed

Zhuo, Yue; Cunningham, Brian T

2015-08-28

We review the development and application of nanostructured photonic crystal surfaces and a hyperspectral reflectance imaging detection instrument which, when used together, represent a new form of optical microscopy that enables label-free, quantitative, and kinetic monitoring of biomaterial interaction with substrate surfaces. Photonic Crystal Enhanced Microscopy (PCEM) has been used to detect broad classes of materials which include dielectric nanoparticles, metal plasmonic nanoparticles, biomolecular layers, and live cells. Because PCEM does not require cytotoxic stains or photobleachable fluorescent dyes, it is especially useful for monitoring the long-term interactions of cells with extracellular matrix surfaces. PCEM is only sensitive to the attachment of cell components within ~200 nm of the photonic crystal surface, which may correspond to the region of most interest for adhesion processes that involve stem cell differentiation, chemotaxis, and metastasis. PCEM has also demonstrated sufficient sensitivity for sensing nanoparticle contrast agents that are roughly the same size as protein molecules, which may enable applications in "digital" diagnostics with single molecule sensing resolution. We will review PCEM's development history, operating principles, nanostructure design, and imaging modalities that enable tracking of optical scatterers, emitters, absorbers, and centers of dielectric permittivity.

Label-Free Biosensor Imaging on Photonic Crystal Surfaces

PubMed Central

Zhuo, Yue; Cunningham, Brian T.

2015-01-01

We review the development and application of nanostructured photonic crystal surfaces and a hyperspectral reflectance imaging detection instrument which, when used together, represent a new form of optical microscopy that enables label-free, quantitative, and kinetic monitoring of biomaterial interaction with substrate surfaces. Photonic Crystal Enhanced Microscopy (PCEM) has been used to detect broad classes of materials which include dielectric nanoparticles, metal plasmonic nanoparticles, biomolecular layers, and live cells. Because PCEM does not require cytotoxic stains or photobleachable fluorescent dyes, it is especially useful for monitoring the long-term interactions of cells with extracellular matrix surfaces. PCEM is only sensitive to the attachment of cell components within ~200 nm of the photonic crystal surface, which may correspond to the region of most interest for adhesion processes that involve stem cell differentiation, chemotaxis, and metastasis. PCEM has also demonstrated sufficient sensitivity for sensing nanoparticle contrast agents that are roughly the same size as protein molecules, which may enable applications in “digital” diagnostics with single molecule sensing resolution. We will review PCEM’s development history, operating principles, nanostructure design, and imaging modalities that enable tracking of optical scatterers, emitters, absorbers, and centers of dielectric permittivity. PMID:26343684

Guided-Wave Optical Biosensors

PubMed Central

Passaro, Vittorio M. N.; Dell'Olio, Francesco; Casamassima, Biagio; De Leonardis, Francesco

2007-01-01

Guided-wave optical biosensors are reviewed in this paper. Advantages related to optical technologies are presented and integrated architectures are investigated in detail. Main classes of bio receptors and the most attractive optical transduction mechanisms are discussed. The possibility to use Mach-Zehnder and Young interferometers, microdisk and microring resonators, surface plasmon resonance, hollow and antiresonant waveguides, and Bragg gratings to realize very sensitive and selective, ultra-compact and fast biosensors is discussed. Finally, CMOS-compatible technologies are proved to be the most attractive for fabrication of guided-wave photonic biosensors.

Introduction to biosensors.

PubMed

Bhalla, Nikhil; Jolly, Pawan; Formisano, Nello; Estrela, Pedro

2016-06-30

Biosensors are nowadays ubiquitous in biomedical diagnosis as well as a wide range of other areas such as point-of-care monitoring of treatment and disease progression, environmental monitoring, food control, drug discovery, forensics and biomedical research. A wide range of techniques can be used for the development of biosensors. Their coupling with high-affinity biomolecules allows the sensitive and selective detection of a range of analytes. We give a general introduction to biosensors and biosensing technologies, including a brief historical overview, introducing key developments in the field and illustrating the breadth of biomolecular sensing strategies and the expansion of nanotechnological approaches that are now available. © 2016 The Author(s). Published by Portland Press Limited on behalf of the Biochemical Society.

Development of quantum dots-based biosensor towards on-farm detection of subclinical ketosis.

PubMed

Weng, Xuan; Chen, Longyan; Neethirajan, Suresh; Duffield, Todd

2015-10-15

Early detection of dairy animal health issues allows the producer or veterinarian to intervene before the animals' production levels, or even survival, is threatened. An increased concentration of β-hydroxybutyrate (βHBA) is a key biomarker for diagnosis of subclinical ketosis (SCK), and provides information on the health stress in cows well before any external symptoms are observable. In this study, quantum dots (QDs) modified with cofactor nicotinamide adenine dinucleotide (NAD(+)) were prepared for the sensing event, by which the βHBA concentration in the cow's blood and milk samples was determined via fluorescence analysis of the functionalized QDs. The detection was performed on a custom designed microfluidic platform combining with a low cost and miniaturized optical sensor. The sensing mechanism was first validated by a microplate reader method and then applied to the microfluidic platform. Standard βHBA solution, βHBA in blood and milk samples from cows were successfully measured by this novel technology with a detection limit at a level of 35 µM. Side by side comparison of the developed microfluidic biosensor with a commercial kit presented its good performance. Copyright © 2015 Elsevier B.V. All rights reserved.

Biosensors-on-chip: a topical review

NASA Astrophysics Data System (ADS)

Chen, Sensen; Shamsi, Mohtashim H.

2017-08-01

This review will examine the integration of two fields that are currently at the forefront of science, i.e. biosensors and microfluidics. As a lab-on-a-chip (LOC) technology, microfluidics has been enriched by the integration of various detection tools for analyte detection and quantitation. The application of such microfluidic platforms is greatly increased in the area of biosensors geared towards point-of-care diagnostics. Together, the merger of microfluidics and biosensors has generated miniaturized devices for sample processing and sensitive detection with quantitation. We believe that microfluidic biosensors (biosensors-on-chip) are essential for developing robust and cost effective point-of-care diagnostics. This review is relevant to a variety of disciplines, such as medical science, clinical diagnostics, LOC technologies including MEMs/NEMs, and analytical science. Specifically, this review will appeal to scientists working in the two overlapping fields of biosensors and microfluidics, and will also help new scientists to find their directions in developing point-of-care devices.

Biosensors in Clinical Practice: Focus on Oncohematology

PubMed Central

Fracchiolla, Nicola S.; Artuso, Silvia; Cortelezzi, Agostino

2013-01-01

Biosensors are devices that are capable of detecting specific biological analytes and converting their presence or concentration into some electrical, thermal, optical or other signal that can be easily analysed. The first biosensor was designed by Clark and Lyons in 1962 as a means of measuring glucose. Since then, much progress has been made and the applications of biosensors are today potentially boundless. This review is limited to their clinical applications, particularly in the field of oncohematology. Biosensors have recently been developed in order to improve the diagnosis and treatment of patients affected by hematological malignancies, such as the biosensor for assessing the in vitro pre-treatment efficacy of cytarabine in acute myeloid leukemia, and the fluorescence resonance energy transfer-based biosensor for assessing the efficacy of imatinib in chronic myeloid leukemia. The review also considers the challenges and future perspectives of biosensors in clinical practice. PMID:23673681

Ultrasensitive and high-throughput analysis of chlorophyll a in marine phytoplankton extracts using a fluorescence microplate reader.

PubMed

Mandalakis, Manolis; Stravinskaitė, Austėja; Lagaria, Anna; Psarra, Stella; Polymenakou, Paraskevi

2017-07-01

Chlorophyll a (Chl a) is the predominant pigment in every single photosynthesizing organism including phytoplankton and one of the most commonly measured water quality parameters. Various methods are available for Chl a analysis, but the majority of them are of limited throughput and require considerable effort and time from the operator. The present study describes a high-throughput, microplate-based fluorometric assay for rapid quantification of Chl a in phytoplankton extracts. Microplate sealing combined with ice cooling was proved an effective means for diminishing solvent evaporation during sample loading and minimized the analytical errors involved in Chl a measurements with a fluorescence microplate reader. A set of operating parameters (settling time, detector gain, sample volume) were also optimized to further improve the intensity and reproducibility of Chl a fluorescence signal. A quadratic regression model provided the best fit (r 2  = 0.9998) across the entire calibration range (0.05-240 pg μL -1 ). The method offered excellent intra- and interday precision (% RSD 2.2 to 11.2%) and accuracy (% relative error -3.8 to 13.8%), while it presented particularly low limits of detection (0.044 pg μL -1 ) and quantification (0.132 pg μL -1 ). The present assay was successfully applied on marine phytoplankton extracts, and the overall results were consistent (average % relative error -14.8%) with Chl a concentrations (including divinyl Chl a) measured by high-performance liquid chromatography (HPLC). More importantly, the microplate-based method allowed the analysis of 96 samples/standards within a few minutes, instead of hours or days, when using a traditional cuvette-based fluorometer or an HPLC system. Graphical abstract TChl a concentrations (i.e. sum of Chl a and divinyl Chl a in ng L -1 ) measured in seawater samples by HPLC and fluorescence microplate reader.

Structural Color Patterns by Electrohydrodynamic Jet Printed Photonic Crystals.

PubMed

Ding, Haibo; Zhu, Cun; Tian, Lei; Liu, Cihui; Fu, Guangbin; Shang, Luoran; Gu, Zhongze

2017-04-05

In this work, we demonstrate the fabrication of photonic crystal patterns with controllable morphologies and structural colors utilizing electrohydrodynamic jet (E-jet) printing with colloidal crystal inks. The final shape of photonic crystal units is controlled by the applied voltage signal and wettability of the substrate. Optical properties of the structural color patterns are tuned by the self-assembly of the silica nanoparticle building blocks. Using this direct printing technique, it is feasible to print customized functional patterns composed of photonic crystal dots or photonic crystal lines according to relevant printing mode and predesigned tracks. This is the first report for E-jet printing with colloidal crystal inks. Our results exhibit promising applications in displays, biosensors, and other functional devices.

The use of a single titanium microplate in displaced pediatric parasymphysial mandibular fractures.

PubMed

Abdullah, Walid A

2009-07-01

The objective of this study was to evaluate the use of one titanium microplate in the fixation of displaced pediatric parasymphysial mandibular fractures. The study was conducted on 7 children in the mixed dentition stage with displaced parasymphysial fracture. Patients' age ranged between 5 years 9 months and 8 years 4 months with an average of 7 years 1 month. Fractured bone segments were exposed, reduced and then fixed using 1.5 linear microplates at the inferior border of the mandible using monocortical screws, with 1.5 mm in diameter and 5 mm in length. Stainless steel wire was used as a tension band by ligating the teeth around the fracture line. Patients were followed up for occlusion and stability clinically and radiographically (panoramic X-ray and CT). According to clinical and radiographic post-operative follow-up, none of the patients showed displacement of the fixed bony segments. The present study concluded that using one microplate with 1.5 monocortical microscrews and dental tension band by a stainless steel wire could be adequate for fixing displaced pediatric parasymphysial mandibular fractures. This technique has the following advantages: decreases the amount of titanium used, decreases the risk of injury of the roots and teeth buds, and decreases the cost and time of surgery.

The use of a single titanium microplate in displaced pediatric parasymphysial mandibular fractures

PubMed Central

Abdullah, Walid A.

2009-01-01

Objective The objective of this study was to evaluate the use of one titanium microplate in the fixation of displaced pediatric parasymphysial mandibular fractures. Materials and methods The study was conducted on 7 children in the mixed dentition stage with displaced parasymphysial fracture. Patients’ age ranged between 5 years 9 months and 8 years 4 months with an average of 7 years 1 month. Fractured bone segments were exposed, reduced and then fixed using 1.5 linear microplates at the inferior border of the mandible using monocortical screws, with 1.5 mm in diameter and 5 mm in length. Stainless steel wire was used as a tension band by ligating the teeth around the fracture line. Patients were followed up for occlusion and stability clinically and radiographically (panoramic X-ray and CT). Results According to clinical and radiographic post-operative follow-up, none of the patients showed displacement of the fixed bony segments. Conclusion The present study concluded that using one microplate with 1.5 monocortical microscrews and dental tension band by a stainless steel wire could be adequate for fixing displaced pediatric parasymphysial mandibular fractures. This technique has the following advantages: decreases the amount of titanium used, decreases the risk of injury of the roots and teeth buds, and decreases the cost and time of surgery. PMID:23960466

Triple Junctions, Boninites, and a New Microplate in the Western Pacific

NASA Astrophysics Data System (ADS)

Flores, J. A.; Casey, J.

2017-12-01

A new microplate has been discovered while trying to correlate melting processes in subduction zones that are forming boninites along the southern Mariana Plate. The westward boundary between the Mariana plate and the Philippine Sea plate is along a well-defined back-arc spreading center. The southern extension of this spreading center to the intersection with the Mariana Trench does not have a recognized morphological boundary. Previous work has hypothesized that subduction beneath a spreading center provides conditions required for boninite petrogenesis. Therefore, the exact location of the trench-trench-ridge triple junction needs to be found and correlated with known boninite locations. The triple junction was found using fault plane solutions to constrain the southern boundary of the two plates as it transects across the forearc. Normal faults suggest the triple junction to be at approximately 11.9N 144.1W; slip direction of reverse faults associated with the subducting plate are dominantly north-south west of this junction and northwest-southeast on the east side. While locating the southern boundary, the nucleation of a new spreading center that creates a ridge-ridge-ridge triple junction was found. The main spreading center trends mostly north-south until about 12.5N 143W, where two other spreading centers meet. The western spreading zone trends mostly east-west and seems to be in its infancy whereas there is another spreading center trending northwest-southeast. It is this last spreading center that forms the trench-ridge-trench triple junction. Discovery of these triple junctions isolates a piece of lithosphere that we interpret to be a new microplate that we name the Challenger Microplate.

Zinc oxide nano-rods based glucose biosensor devices fabrication

NASA Astrophysics Data System (ADS)

Wahab, H. A.; Salama, A. A.; El Saeid, A. A.; Willander, M.; Nur, O.; Battisha, I. K.

2018-06-01

ZnO is distinguished multifunctional material that has wide applications in biochemical sensor devices. For extracellular measurements, Zinc oxide nano-rods will be deposited on conducting plastic substrate with annealing temperature 150 °C (ZNRP150) and silver wire with annealing temperature 250 °C (ZNRW250), for the extracellular glucose concentration determination with functionalized ZNR-coated biosensors. It was performed in phosphate buffer saline (PBS) over the range from 1 μM to 10 mM and on human blood plasma. The prepared samples crystal structure and surface morphologies were characterized by XRD and field emission scanning electron microscope FESEM respectively.

A New Laccase Based Biosensor for Tartrazine.

PubMed

Mazlan, Siti Zulaikha; Lee, Yook Heng; Hanifah, Sharina Abu

2017-12-09

Laccase enzyme, a commonly used enzyme for the construction of biosensors for phenolic compounds was used for the first time to develop a new biosensor for the determination of the azo-dye tartrazine. The electrochemical biosensor was based on the immobilization of laccase on functionalized methacrylate-acrylate microspheres. The biosensor membrane is a composite of the laccase conjugated microspheres and gold nanoparticles (AuNPs) coated on a carbon-paste screen-printed electrode. The reaction involving tartrazine can be catalyzed by laccase enzyme, where the current change was measured by differential pulse voltammetry (DPV) at 1.1 V. The anodic peak current was linear within the tartrazine concentration range of 0.2 to 14 μM ( R ² = 0.979) and the detection limit was 0.04 μM. Common food ingredients or additives such as glucose, sucrose, ascorbic acid, phenol and sunset yellow did not interfere with the biosensor response. Furthermore, the biosensor response was stable up to 30 days of storage period at 4 °C. Foods and beverage were used as real samples for the biosensor validation. The biosensor response to tartrazine showed no significant difference with a standard HPLC method for tartrazine analysis.

A New Laccase Based Biosensor for Tartrazine

PubMed Central

Mazlan, Siti Zulaikha; Lee, Yook Heng; Hanifah, Sharina Abu

2017-01-01

Laccase enzyme, a commonly used enzyme for the construction of biosensors for phenolic compounds was used for the first time to develop a new biosensor for the determination of the azo-dye tartrazine. The electrochemical biosensor was based on the immobilization of laccase on functionalized methacrylate-acrylate microspheres. The biosensor membrane is a composite of the laccase conjugated microspheres and gold nanoparticles (AuNPs) coated on a carbon-paste screen-printed electrode. The reaction involving tartrazine can be catalyzed by laccase enzyme, where the current change was measured by differential pulse voltammetry (DPV) at 1.1 V. The anodic peak current was linear within the tartrazine concentration range of 0.2 to 14 μM (R2 = 0.979) and the detection limit was 0.04 μM. Common food ingredients or additives such as glucose, sucrose, ascorbic acid, phenol and sunset yellow did not interfere with the biosensor response. Furthermore, the biosensor response was stable up to 30 days of storage period at 4 °C. Foods and beverage were used as real samples for the biosensor validation. The biosensor response to tartrazine showed no significant difference with a standard HPLC method for tartrazine analysis. PMID:29232842

Biosensor for metal analysis and speciation

DOEpatents

Aiken, Abigail M.; Peyton, Brent M.; Apel, William A.; Petersen, James N.

2007-01-30

A biosensor for metal analysis and speciation is disclosed. The biosensor comprises an electron carrier immobilized to a surface of an electrode and a layer of an immobilized enzyme adjacent to the electrode. The immobilized enzyme comprises an enzyme having biological activity inhibited by a metal to be detected by the biosensor.

Liquid crystal-based glucose biosensor functionalized with mixed PAA and QP4VP brushes.

PubMed

Khan, Mashooq; Park, Soo-Young

2015-06-15

4-Cyano-4'-pentylbiphenyl (5CB) in a transmission electron microscopy (TEM) grid was developed for glucose detection by coating with a monolayer of mixed polymer brushes using poly(acrylicacid-b-4-cynobiphenyl-4'-oxyundecylacrylate) (PAA-b-LCP) and quaternized poly(4-vinylpyridine-b-4-cynobiphenyl-4'-oxyundecylacrylate) (QP4VP-b-LCP) (LCP stands for liquid crystal polymer) at the 5CB/aqueous interface. The resultant 5CB in TEM grid was functionalized with the PAA and QP4VP brushes, which were strongly anchored by the LCP block. The PAA brush rendered the 5CB/aqueous interface pH-responsive and the QP4VP brush immobilized glucose oxidase (GOx) through electrostatic interactions without the aid of coupling agents. The glucose was detected through a homeotropic-to-planar orientational transition of the 5CB observed through a polarized optical microscope (POM) under crossed polarizers. The optimum immobilization with a 0.78 µM GOx solution on the dual-brush-coated TEM grid enabled glucose detection at concentrations higher than 0.5 mM with response times shorter than 180 s. This TEM grid glucose sensor provided a linear response of birefringence of the 5CB to glucose concentrations ranging from 0.5 to 11 mM with a Michaelis-Menten constant (Km) of 1.67 mM. This new and sensitive glucose biosensor has the advantages of low production cost, simple enzyme immobilization, high enzyme sensitivity and stability, and easy detection with POM, and may be useful for prescreening the glucose level in the human body. Copyright © 2015 Elsevier B.V. All rights reserved.

Hybridization assay of insect antifreezing protein gene by novel multilayered porous silicon nucleic acid biosensor.

PubMed

Lv, Xiaoyi; Chen, Liangliang; Zhang, Hongyan; Mo, Jiaqing; Zhong, Furu; Lv, Changwu; Ma, Ji; Jia, Zhenhong

2013-01-15

A fabrication of a novel simple porous silicon polybasic photonic crystal with symmetrical structure has been reported as a nucleic acid biosensor for detecting antifreeze protein gene in insects (Microdera puntipennis dzhungarica), which would be helpful in the development of some new transgenic plants with tolerance of freezing stress. Compared to various porous silicon-based photonic configurations, porous silicon polytype layered structure is quite easy to prepare and shows more stability; moreover, polybasic photonic crystals with symmetrical structure exhibit interesting optical properties with a sharp resonance in the reflectance spectrum, giving a higher Q factor which causes higher sensitivity for sensing performance. In this experiment, DNA oligonucleotides were immobilized into the porous silicon pores using a standard crosslink chemistry method. The porous silicon polybasic symmetrical structure sensor possesses high specificity in performing controlled experiments with non-complementary DNA. The detection limit was found to be 21.3nM for DNA oligonucleotides. The fabricated multilayered porous silicon-based DNA biosensor has potential commercial applications in clinical chemistry for determination of an antifreeze protein gene or other genes. Copyright © 2012 Elsevier B.V. All rights reserved.

Electronic Biosensors Based on III-Nitride Semiconductors.

PubMed

Kirste, Ronny; Rohrbaugh, Nathaniel; Bryan, Isaac; Bryan, Zachary; Collazo, Ramon; Ivanisevic, Albena

2015-01-01

We review recent advances of AlGaN/GaN high-electron-mobility transistor (HEMT)-based electronic biosensors. We discuss properties and fabrication of III-nitride-based biosensors. Because of their superior biocompatibility and aqueous stability, GaN-based devices are ready to be implemented as next-generation biosensors. We review surface properties, cleaning, and passivation as well as different pathways toward functionalization, and critically analyze III-nitride-based biosensors demonstrated in the literature, including those detecting DNA, bacteria, cancer antibodies, and toxins. We also discuss the high potential of these biosensors for monitoring living cardiac, fibroblast, and nerve cells. Finally, we report on current developments of covalent chemical functionalization of III-nitride devices. Our review concludes with a short outlook on future challenges and projected implementation directions of GaN-based HEMT biosensors.

Nanoshell-Enhanced Raman Spectroscopy on a Microplate for Staphylococcal Enterotoxin B Sensing.

PubMed

Wang, Wenbin; Wang, Weiwei; Liu, Liqiang; Xu, Liguang; Kuang, Hua; Zhu, Jianping; Xu, Chuanlai

2016-06-22

A sensitive surface-enhanced Raman scattering (SERS) immunosensor based on the Au nanoparticle (Au NP) shell structure was developed to detect staphylococcal enterotoxin B (SEB) on a microplate. Au NPs modified with 4-nitrothiophenol (4-NTP) and coated with Ag shell of controlled thickness at 6.6 nm exhibited excellent SERS intensity and were used as signal reporters in the detection of SEB. The engaged 4-NTP allowed the significant electromagnetic enhancement between Au NPs and the Ag shell and prevented the dissociation of the Raman reporter. More importantly, 4-NTP-differentiated SERS signals between the sample and microplate. The SERS-based immunosensor had a limit of detection of 1.3 pg/mL SEB. Analysis of SEB-spiked milk samples revealed that the developed method had high accuracy. Therefore, the SERS-encoded Au@Ag core-shell structure-based immunosensor is promising for the detection of biotoxins, pathogens, and environmental pollutants.

S-Layer Protein-Based Biosensors.

PubMed

Schuster, Bernhard

2018-04-11

The present paper highlights the application of bacterial surface (S-) layer proteins as versatile components for the fabrication of biosensors. One technologically relevant feature of S-layer proteins is their ability to self-assemble on many surfaces and interfaces to form a crystalline two-dimensional (2D) protein lattice. The S-layer lattice on the surface of a biosensor becomes part of the interface architecture linking the bioreceptor to the transducer interface, which may cause signal amplification. The S-layer lattice as ultrathin, highly porous structure with functional groups in a well-defined special distribution and orientation and an overall anti-fouling characteristics can significantly raise the limit in terms of variety and the ease of bioreceptor immobilization, compactness of bioreceptor molecule arrangement, sensitivity, specificity, and detection limit for many types of biosensors. The present paper discusses and summarizes examples for the successful implementation of S-layer lattices on biosensor surfaces in order to give a comprehensive overview on the application potential of these bioinspired S-layer protein-based biosensors.

Effects of van der Waals Force and Thermal Stresses on Pull-in Instability of Clamped Rectangular Microplates.

PubMed

Batra, Romesh C; Porfiri, Maurizio; Spinello, Davide

2008-02-15

We study the influence of von Karman nonlinearity, van der Waals force, and a athermal stresses on pull-in instability and small vibrations of electrostatically actuated mi-croplates. We use the Galerkin method to develop a tractable reduced-order model for elec-trostatically actuated clamped rectangular microplates in the presence of van der Waals forcesand thermal stresses. More specifically, we reduce the governing two-dimensional nonlineartransient boundary-value problem to a single nonlinear ordinary differential equation. For thestatic problem, the pull-in voltage and the pull-in displacement are determined by solving apair of nonlinear algebraic equations. The fundamental vibration frequency corresponding toa deflected configuration of the microplate is determined by solving a linear algebraic equa-tion. The proposed reduced-order model allows for accurately estimating the combined effectsof van der Waals force and thermal stresses on the pull-in voltage and the pull-in deflectionprofile with an extremely limited computational effort.

Effects of van der Waals Force and Thermal Stresses on Pull-in Instability of Clamped Rectangular Microplates

PubMed Central

Batra, Romesh C.; Porfiri, Maurizio; Spinello, Davide

2008-01-01

We study the influence of von Kármán nonlinearity, van der Waals force, and thermal stresses on pull-in instability and small vibrations of electrostatically actuated microplates. We use the Galerkin method to develop a tractable reduced-order model for electrostatically actuated clamped rectangular microplates in the presence of van der Waals forces and thermal stresses. More specifically, we reduce the governing two-dimensional nonlinear transient boundary-value problem to a single nonlinear ordinary differential equation. For the static problem, the pull-in voltage and the pull-in displacement are determined by solving a pair of nonlinear algebraic equations. The fundamental vibration frequency corresponding to a deflected configuration of the microplate is determined by solving a linear algebraic equation. The proposed reduced-order model allows for accurately estimating the combined effects of van der Waals force and thermal stresses on the pull-in voltage and the pull-in deflection profile with an extremely limited computational effort. PMID:27879752

System-level integration of active silicon photonic biosensors

NASA Astrophysics Data System (ADS)

Laplatine, L.; Al'Mrayat, O.; Luan, E.; Fang, C.; Rezaiezadeh, S.; Ratner, D. M.; Cheung, K.; Dattner, Y.; Chrostowski, L.

2017-02-01

Biosensors based on silicon photonic integrated circuits have attracted a growing interest in recent years. The use of sub-micron silicon waveguides to propagate near-infrared light allows for the drastic reduction of the optical system size, while increasing its complexity and sensitivity. Using silicon as the propagating medium also leverages the fabrication capabilities of CMOS foundries, which offer low-cost mass production. Researchers have deeply investigated photonic sensor devices, such as ring resonators, interferometers and photonic crystals, but the practical integration of silicon photonic biochips as part of a complete system has received less attention. Herein, we present a practical system-level architecture which can be employed to integrate the aforementioned photonic biosensors. We describe a system based on 1 mm2 dies that integrate germanium photodetectors and a single light coupling device. The die are embedded into a 16x16 mm2 epoxy package to enable microfluidic and electrical integration. First, we demonstrate a simple process to mimic Fan-Out Wafer-level-Packaging, which enables low-cost mass production. We then characterize the photodetectors in the photovoltaic mode, which exhibit high sensitivity at low optical power. Finally, we present a new grating coupler concept to relax the lateral alignment tolerance down to +/- 50 μm at 1-dB (80%) power penalty, which should permit non-experts to use the biochips in a"plug-and-play" style. The system-level integration demonstrated in this study paves the way towards the mass production of low-cost and highly sensitive biosensors, and can facilitate their wide adoption for biomedical and agro-environmental applications.

Using Isolated Mitochondria from Minimal Quantities of Mouse Skeletal Muscle for High throughput Microplate Respiratory Measurements.

PubMed

Boutagy, Nabil E; Rogers, George W; Pyne, Emily S; Ali, Mostafa M; Hulver, Matthew W; Frisard, Madlyn I

2015-10-30

Skeletal muscle mitochondria play a specific role in many disease pathologies. As such, the measurement of oxygen consumption as an indicator of mitochondrial function in this tissue has become more prevalent. Although many technologies and assays exist that measure mitochondrial respiratory pathways in a variety of cells, tissue and species, there is currently a void in the literature in regards to the compilation of these assays using isolated mitochondria from mouse skeletal muscle for use in microplate based technologies. Importantly, the use of microplate based respirometric assays is growing among mitochondrial biologists as it allows for high throughput measurements using minimal quantities of isolated mitochondria. Therefore, a collection of microplate based respirometric assays were developed that are able to assess mechanistic changes/adaptations in oxygen consumption in a commonly used animal model. The methods presented herein provide step-by-step instructions to perform these assays with an optimal amount of mitochondrial protein and reagents, and high precision as evidenced by the minimal variance across the dynamic range of each assay.

HNO₃-assisted polyol synthesis of ultralarge single-crystalline Ag microplates and their far propagation length of surface plasmon polariton.

PubMed

Chang, Cheng-Wei; Lin, Fan-Cheng; Chiu, Chun-Ya; Su, Chung-Yi; Huang, Jer-Shing; Perng, Tsong-Pyng; Yen, Ta-Jen

2014-07-23

We developed a HNO3-assisted polyol reduction method to synthesize ultralarge single-crystalline Ag microplates routinely. The edge length of the synthesized Ag microplates reaches 50 μm, and their top facets are (111). The mechanism for dramatically enlarging single-crystalline Ag structure stems from a series of competitive anisotropic growths, primarily governed by carefully tuning the adsorption of Ag(0) by ethylene glycol and the desorption of Ag(0) by a cyanide ion on Ag(100). Finally, we measured the propagation length of surface plasmon polaritons along the air/Ag interface under 534 nm laser excitation. Our single-crystalline Ag microplate exhibited a propagation length (11.22 μm) considerably greater than that of the conventional E-gun deposited Ag thin film (5.27 μm).

Angle-resolved diffraction grating biosensor based on porous silicon

DOE Office of Scientific and Technical Information (OSTI.GOV)

Lv, Changwu; Li, Peng; Jia, Zhenhong, E-mail: jzhh@xju.edu.cn

2016-03-07

In this study, an optical biosensor based on a porous silicon composite structure was fabricated using a simple method. This structure consists of a thin, porous silicon surface diffraction grating and a one-dimensional porous silicon photonic crystal. An angle-resolved diffraction efficiency spectrum was obtained by measuring the diffraction efficiency at a range of incident angles. The angle-resolved diffraction efficiency of the 2nd and 3rd orders was studied experimentally and theoretically. The device was sensitive to the change of refractive index in the presence of a biomolecule indicated by the shift of the diffraction efficiency spectrum. The sensitivity of this sensormore » was investigated through use of an 8 base pair antifreeze protein DNA hybridization. The shifts of the angle-resolved diffraction efficiency spectrum showed a relationship with the change of the refractive index, and the detection limit of the biosensor reached 41.7 nM. This optical device is highly sensitive, inexpensive, and simple to fabricate. Using shifts in diffraction efficiency spectrum to detect biological molecules has not yet been explored, so this study establishes a foundation for future work.« less

Correction of Microplate Data from High-Throughput Screening.

PubMed

Wang, Yuhong; Huang, Ruili

2016-01-01

High-throughput screening (HTS) makes it possible to collect cellular response data from a large number of cell lines and small molecules in a timely and cost-effective manner. The errors and noises in the microplate-formatted data from HTS have unique characteristics, and they can be generally grouped into three categories: run-wise (temporal, multiple plates), plate-wise (background pattern, single plate), and well-wise (single well). In this chapter, we describe a systematic solution for identifying and correcting such errors and noises, mainly basing on pattern recognition and digital signal processing technologies.

Progress of new label-free techniques for biosensors: a review.

PubMed

Sang, Shengbo; Wang, Yajun; Feng, Qiliang; Wei, Ye; Ji, Jianlong; Zhang, Wendong

2016-01-01

The detection techniques used in biosensors can be broadly classified into label-based and label-free. Label-based detection relies on the specific properties of labels for detecting a particular target. In contrast, label-free detection is suitable for the target molecules that are not labeled or the screening of analytes which are not easy to tag. Also, more types of label-free biosensors have emerged with developments in biotechnology. The latest developed techniques in label-free biosensors, such as field-effect transistors-based biosensors including carbon nanotube field-effect transistor biosensors, graphene field-effect transistor biosensors and silicon nanowire field-effect transistor biosensors, magnetoelastic biosensors, optical-based biosensors, surface stress-based biosensors and other type of biosensors based on the nanotechnology are discussed. The sensing principles, configurations, sensing performance, applications, advantages and restriction of different label-free based biosensors are considered and discussed in this review. Most concepts included in this survey could certainly be applied to the development of this kind of biosensor in the future.

Evaluation of the Alexon-Trend ProSpecT Campylobacter Microplate Assay

PubMed Central

Tolcin, Rita; LaSalvia, Margaret M.; Kirkley, Barbara A.; Vetter, Emily A.; Cockerill, Franklin R.; Procop, Gary W.

2000-01-01

We evaluated stool specimens known to contain or be free of Campylobacter by traditional culture, using the ProSpecT Campylobacter microplate assay (Alexon-Trend, Ramsey, Minn.). This rapid enzyme immunoassay for the detection of Campylobacter-specific antigens demonstrated 96% sensitivity and 99% specificity and is an acceptable alternative method of Campylobacter detection. PMID:11015419

Modeling of mixing in 96-well microplates observed with fluorescence indicators.

PubMed

Weiss, Svenja; John, Gernot T; Klimant, Ingo; Heinzle, Elmar

2002-01-01

Mixing in 96-well microplates was studied using soluble pH indicators and a fluorescence pH sensor. Small amounts of alkali were added with the aid of a multichannel pipet, a piston pump, and a piezoelectric actuator. Mixing patterns were observed visually using a video camera. Addition of drops each of about 1 nL with the piezoelectric actuator resulted in umbrella and double-disklike shapes. Convective mixing was mainly observed in the upper part of the well, whereas the lower part was only mixed quickly when using the multichannel pipet and the piston pump with an addition volume of 5 microL or larger. Estimated mixing times were between a few seconds and several minutes. Mixing by liquid dispensing was much more effective than by shaking. A mixing model consisting of 21 elements could describe mixing dynamics observed by the dissolved fluorescence dye and by the optical immobilized pH sensor. This model can be applied for designing pH control in microplates or for design of kinetic experiments with liquid addition.

Recent Trends in Biosensors

NASA Astrophysics Data System (ADS)

Karube, Isao

The determination of organic compounds in foods is very important in food industries. A various compounds are contained in foods, selective determination methods are required for food processing and analysis. Electrochemical monitoring devices (biosensors) employing immobilized biocatalysts such as immobilized enzymes, organelles, microorganisms, and tissue have definite advantages. The enzyme Sensors consisted of immobilized enzymes and electrochemical devices. Enzyme sensors could be used for the determination of sugars, amino acids, organic acids, alcohols, lipids, nucleic acid derivatives, etc.. Furthermore, a multifunctional biosensor for the determination of several compounds has been developed for food processing. On the other hand, microbial sensors consisted of immobilized microorganisms and electrodes have been used for industrial and environmental analysis. Microbial sensors were applied for the determination of sugars, organic acids, alcohols, amino acids, mutagens, me thane, ammonia, and BOD. Furthermore, micro-biosensors using immobilized biocatalysts and ion sensitive field effect transistor or microelectrodes prepared by silicon fabrication technologies have been developed for medical ap. plication and food processing. This review summarizes the design and application of biosensors.

A methodological combined framework for roadmapping biosensor research: a fault tree analysis approach within a strategic technology evaluation frame.

PubMed

Siontorou, Christina G; Batzias, Fragiskos A

2014-03-01

Biosensor technology began in the 1960s to revolutionize instrumentation and measurement. Despite the glucose sensor market success that revolutionized medical diagnostics, and artificial pancreas promise currently the approval stage, the industry is reluctant to capitalize on other relevant university-produced knowledge and innovation. On the other hand, the scientific literature is extensive and persisting, while the number of university-hosted biosensor groups is growing. Considering the limited marketability of biosensors compared to the available research output, the biosensor field has been used by the present authors as a suitable paradigm for developing a methodological combined framework for "roadmapping" university research output in this discipline. This framework adopts the basic principles of the Analytic Hierarchy Process (AHP), replacing the lower level of technology alternatives with internal barriers (drawbacks, limitations, disadvantages), modeled through fault tree analysis (FTA) relying on fuzzy reasoning to count for uncertainty. The proposed methodology is validated retrospectively using ion selective field effect transistor (ISFET) - based biosensors as a case example, and then implemented prospectively membrane biosensors, putting an emphasis on the manufacturability issues. The analysis performed the trajectory of membrane platforms differently than the available market roadmaps that, considering the vast industrial experience in tailoring and handling crystallic forms, suggest the technology path of biomimetic and synthetic materials. The results presented herein indicate that future trajectories lie along with nanotechnology, and especially nanofabrication and nano-bioinformatics, and focused, more on the science-path, that is, on controlling the natural process of self-assembly and the thermodynamics of bioelement-lipid interaction. This retained the nature-derived sensitivity of the biosensor platform, pointing out the differences

Biosensoric potential of microbial fuel cells.

PubMed

Schneider, György; Kovács, Tamás; Rákhely, Gábor; Czeller, Miklós

2016-08-01

Recent progress in microbial fuel cell (MFC) technology has highlighted the potential of these devices to be used as biosensors. The advantages of MFC-based biosensors are that they are phenotypic and can function in either assay- or flow-through formats. These features make them appropriate for contiguous on-line monitoring in laboratories and for in-field applications. The selectivity of an MFC biosensor depends on the applied microorganisms in the anodic compartment where electron transfer (ET) between the artificial surface (anode) and bacterium occurs. This process strongly determines the internal resistance of the sensoric system and thus influences signal outcome and response time. Despite their beneficial characteristics, the number of MFC-based biosensoric applications has been limited until now. The aim of this mini-review is to turn attention to the biosensoric potential of MFCs by summarizing ET mechanisms on which recently established and future sensoric devices are based.

Current Trends in Nanomaterial-Based Amperometric Biosensors

PubMed Central

Hayat, Akhtar; Catanante, Gaëlle; Marty, Jean Louis

2014-01-01

The last decade has witnessed an intensive research effort in the field of electrochemical sensors, with a particular focus on the design of amperometric biosensors for diverse analytical applications. In this context, nanomaterial integration in the construction of amperometric biosensors may constitute one of the most exciting approaches. The attractive properties of nanomaterials have paved the way for the design of a wide variety of biosensors based on various electrochemical detection methods to enhance the analytical characteristics. However, most of these nanostructured materials are not explored in the design of amperometric biosensors. This review aims to provide insight into the diverse properties of nanomaterials that can be possibly explored in the construction of amperometric biosensors. PMID:25494347

Efficient Fluorescence Resonance Energy Transfer between Quantum Dots and Gold Nanoparticles Based on Porous Silicon Photonic Crystal for DNA Detection

PubMed Central

Zhang, Hongyan; Lv, Jie; Jia, Zhenhong

2017-01-01

A novel assembled biosensor was prepared for detecting 16S rRNA, a small-size persistent specific for Actinobacteria. The mechanism of the porous silicon (PS) photonic crystal biosensor is based on the fluorescence resonance energy transfer (FRET) between quantum dots (QDs) and gold nanoparticles (AuNPs) through DNA hybridization, where QDs act as an emission donor and AuNPs serve as a fluorescence quencher. Results showed that the photoluminescence (PL) intensity of PS photonic crystal was drastically increased when the QDs-conjugated probe DNA was adhered to the PS layer by surface modification using a standard cross-link chemistry method. The PL intensity of QDs was decreased when the addition of AuNPs-conjugated complementary 16S rRNA was dropped onto QDs-conjugated PS. Based on the analysis of different target DNA concentration, it was found that the decrease of the PL intensity showed a good linear relationship with complementary DNA concentration in a range from 0.25 to 10 μM, and the detection limit was 328.7 nM. Such an optical FRET biosensor functions on PS-based photonic crystal for DNA detection that differs from the traditional FRET, which is used only in liquid. This method will benefit the development of a new optical FRET label-free biosensor on Si substrate and has great potential in biochips based on integrated optical devices. PMID:28489033

Efficient Fluorescence Resonance Energy Transfer between Quantum Dots and Gold Nanoparticles Based on Porous Silicon Photonic Crystal for DNA Detection.

PubMed

Zhang, Hongyan; Lv, Jie; Jia, Zhenhong

2017-05-10

A novel assembled biosensor was prepared for detecting 16S rRNA, a small-size persistent specific for Actinobacteria. The mechanism of the porous silicon (PS) photonic crystal biosensor is based on the fluorescence resonance energy transfer (FRET) between quantum dots (QDs) and gold nanoparticles (AuNPs) through DNA hybridization, where QDs act as an emission donor and AuNPs serve as a fluorescence quencher. Results showed that the photoluminescence (PL) intensity of PS photonic crystal was drastically increased when the QDs-conjugated probe DNA was adhered to the PS layer by surface modification using a standard cross-link chemistry method. The PL intensity of QDs was decreased when the addition of AuNPs-conjugated complementary 16S rRNA was dropped onto QDs-conjugated PS. Based on the analysis of different target DNA concentration, it was found that the decrease of the PL intensity showed a good linear relationship with complementary DNA concentration in a range from 0.25 to 10 μM, and the detection limit was 328.7 nM. Such an optical FRET biosensor functions on PS-based photonic crystal for DNA detection that differs from the traditional FRET, which is used only in liquid. This method will benefit the development of a new optical FRET label-free biosensor on Si substrate and has great potential in biochips based on integrated optical devices.

Development of electrochemical biosensors with various types of zeolites

NASA Astrophysics Data System (ADS)

Soldatkina, O. V.; Kucherenko, I. S.; Soldatkin, O. O.; Pyeshkova, V. M.; Dudchenko, O. Y.; Akata Kurç, B.; Dzyadevych, S. V.

2018-03-01

In the work, different types of zeolites were used for the development of enzyme-based electrochemical biosensors. Zeolites were added to the biorecognition elements of the biosensors and served as additional components of the biomembranes or adsorbents for enzymes. Three types of biosensors (conductometric, amperometric and potentiometric) were studied. The developed biosensors were compared with the similar biosensors without zeolites. The biosensors contained the following enzymes: urease, glucose oxidase, glutamate oxidase, and acetylcholinesterase and were intended for the detection of urea, glucose, glutamate, and acetylcholine, respectively. Construction of the biosensors using the adsorption of enzymes on zeolites has several advantages: simplicity, good reproducibility, quickness, absence of toxic compounds. These benefits are particularly important for the standardization and further mass production of the biosensors. Furthermore, a biosensor for the sucrose determination contained a three-enzyme system (invertase/mutatorase/glucose oxidase), immobilized by a combination of adsorption on silicalite and cross-linking via glutaraldehyde; such combined immobilization demonstrated better results as compared with adsorption or cross-linking separately. The analysis of urea and sucrose concentrations in the real samples was carried out. The results, obtained with biosensors, had high correlation with the results of traditional analytical methods, thus the developed biosensors are promising for practical applications.

Fabrication of Refractive Index Tunable Polydimethylsiloxane Photonic Crystal for Biosensor Application

NASA Astrophysics Data System (ADS)

Raman, Karthik; Murthy, T. R. Srinivasa; Hegde, G. M.

Photonic crystal based nanostructures are expected to play a significant role in next generation nanophotonic devices. Recent developments in two-dimensional (2D) photonic crystal based devices have created widespread interest as such planar photonic structures are compatible with conventional microelectronic and photonic devices. Various optical components such as waveguides, resonators, modulators and demultiplexers have been designed and fabricated based on 2D photonic crystal geometry. This paper presents the fabrication of refractive index tunable Polydimethylsiloxane (PDMS) polymer based photonic crystals. The advantages of using PDMS are mainly its chemical stability, bio-compatibility and the stack reduces sidewall roughness scattering. The PDMS structure with square lattice was fabricated by using silicon substrate patterned with SU8-2002 resist. The 600 nm period grating of PDMS is then fabricated using Nano-imprinting. In addition, the refractive index of PDMS is modified using certain additive materials. The resulting photonic crystals are suitable for application in photonic integrated circuits and biological applications such as filters, cavities or microlaser waveguides.

Current status of water environment and their microbial biosensor techniques - Part II: Recent trends in microbial biosensor development.

PubMed

Nakamura, Hideaki

2018-05-08

In Part I of the present review series, I presented the current state of the water environment by focusing on Japanese cases and discussed the need to further develop microbial biosensor technologies for the actual water environment. I comprehensively present trends after approximately 2010 in microbial biosensor development for the water environment. In the first section, after briefly summarizing historical studies, recent studies on microbial biosensor principles are introduced. In the second section, recent application studies for the water environment are also introduced. Finally, I conclude the present review series by describing the need to further develop microbial biosensor technologies. Graphical abstract Current water pollution indirectly occurs by anthropogenic eutrophication (Part I). Recent trends in microbial biosensor development for water environment are described in part II of the present review series.

A fluorescence microplate screen assay for the detection of neurite outgrowth and neurotoxicity using an antibody against βIII-tubulin.

PubMed

Popova, Dina; Jacobsson, Stig O P

2014-04-01

The majority of environmental and commercial chemicals have not been evaluated for their potential to cause neurotoxicity. We have investigated if neuron specific anti-βIII-tubulin antibodies are useful in a microplate assay of neurite outgrowth of retinoic acid-induced neurons from mouse P19 embryonal carcinoma cells. By incubating the P19-derived neurons with the primary anti-βIII-tubulin antibody and a secondary Alexa Fluor 488-conjugated antibody, followed by measuring the fluorescence in a microplate reader, a time-dependent increase in anti-βIII-tubulin immunofluorescence was observed. The relative fluorescence units increased by 4.3-fold from 2 to 10 days in culture. The results corresponded well with those obtained by semi-automatic tracing of neurites in fluorescence microscopy images of βIII-tubulin-labeled neurons. The sensitivity of the neurite outgrowth assay using a microplate reader to detect neurotoxicity produced by nocodazole, methyl mercury chloride and okadaic acid was significantly higher than for a cell viability assay measuring intracellular fluorescence of calcein-AM. The microplate-based method to measure toxicity targeting neurites using anti-βIII-tubulin antibodies is however less sensitive than the extracellular lactate dehydrogenase activity assay to detect general cytotoxicity produced by high concentrations of clomipramine, or glutamate-induced excitotoxicity. In conclusion, the fluorescence microplate assay for the detection of neurite outgrowth by measuring changes in βIII-tubulin immunoreactivity is a rapid and sensitive method to assess chemical- or toxin-induced neurite toxicity. Copyright © 2013 Elsevier Ltd. All rights reserved.

Triggered optical biosensor

DOEpatents

Song, Xuedong; Swanson, Basil I.

2001-10-02

An optical biosensor is provided for the detection of a multivalent target biomolecule, the biosensor including a substrate having a bilayer membrane thereon, a recognition molecule situated at the surface, the recognition molecule capable of binding with the multivalent target biomolecule, the recognition molecule further characterized as including a fluorescence label thereon and as being movable at the surface and a device for measuring a fluorescence change in response to binding between the recognition molecule and the multivalent target biomolecule.

In vitro toxicity testing with microplate cell cultures: Impact of cell binding.

PubMed

Gülden, Michael; Schreiner, Jeannine; Seibert, Hasso

2015-06-05

In vitro generated data on toxic potencies are generally based on nominal concentrations. However, cellular and extracellular binding and elimination processes may reduce the available free fraction of a compound. Then, nominal effective concentrations do not represent appropriate measures of toxic exposure in vitro and underestimate toxic potencies. In this study it was investigated whether cell binding can affect the availability of chemicals in microplate based toxicity assays. To this end the cytotoxicity of compounds like mercury chloride, digitonin and alcohol ethoxylates, accumulated by cells via different modes, was investigated in 96-well microplate cultures with varying concentrations of Balb/c 3T3 cells. The median effective nominal concentrations of all but one of the tested compounds depended linearly from the cell concentration. Applying a previously developed equilibrium distribution model cell concentration-independent median effective extracellular concentrations and cell burdens, respectively, could be calculated. The compounds were accumulated by the cells with bioconcentration factors, BCF, between 480 and ≥ 25,000. Cell binding of the alcohol ethoxylates was correlated with their lipophilicity. The results show that significant cell binding can occur even at the small cell volume fractions (∼ 1 × 10(-5) to 3 × 10(-3) L/L) encountered in microplate assays. To what extent cell binding affects the bioavailability depends on the BCF and the cell volume fraction. EC50 measurements in the presence of at least two different cell concentrations allow for excluding or detecting significant cell binding and for determining more appropriate measures of toxic exposure in vitro like median effective extracellular (free) concentrations or cell burdens. Copyright © 2013 Elsevier Ireland Ltd. All rights reserved.

Nanomaterial-Based Electrochemical Biosensors and Bioassays

DOE Office of Scientific and Technical Information (OSTI.GOV)

Liu, Guodong; Mao, Xun; Gurung, Anant

2010-08-31

This book chapter summarizes the recent advance in nanomaterials for electrochemical biosensors and bioassays. Biofunctionalization of nanomaterials for biosensors fabrication and their biomedical applications are discussed.

Synthetic Electric Microbial Biosensors

DTIC Science & Technology

2017-06-10

In particular, monitoring of heavy metals in the environment, drinking water, food , and biological fluids is of interest. Conventional techniques...instances of contamination , and the potential for deliberate spills, interest has grown in portable devices for onsite long-term detection using sensor...biosensor systems for the online detection of a range of contaminants . Synthetic Biology and biosensors Synthetic biology has gained much interest

Portable Microplate Analyzer with a Thermostatic Chamber Based on a Smartphone for On-site Rapid Detection.

PubMed

Wan, Zijian; Zhong, Longjie; Pan, Yuxiang; Li, Hongbo; Zou, Quchao; Su, Kaiqi; Wang, Ping

2017-01-01

A microplate method provides an efficient way to use modern detection technology. However, there are some difficulties concerning on-site detection, such as being non-portable and time-consuming. In this work, a novel portable microplate analyzer with a thermostatic chamber based on a smartphone was designed for rapid on-site detection. An analyzer with a wide-angle lens and an optical filter provides a proper environment for the microplate. A smartphone app-iPlate Monitor was used for RGB analyze of image. After a consistency experiment with a microtiter plate reader (MTPR), the normalized calibration curves were y = 0.7276x + 0.0243 (R 2 = 0.9906) and y = 0.3207x + 0.0094 (R 2 = 0.9917) with a BCA protein kit as well as y = 0.182x + 0.0134 (R 2 = 0.994) and y = 0.0674x + 0.0003 (R 2 = 0.9988) with a glucose kit. The times for obtaining the detection requirement were 15 and 10 min for the BCA protein kit and the glucose kit at 37°C; in contrast, it required more than 30 and 20 min at ambient temperature. Meanwhile, it also showed good repeatability for detections.

One microplate - three orogens: Alps, Dinarides, Apennines and the role of the Adriatic plate

NASA Astrophysics Data System (ADS)

Ustaszewski, Kamil; Le Breton, Eline; Balling, Philipp; Handy, Mark R.; Molli, Giancarlo; Tomljenović, Bruno

2017-04-01

The motion of the Adriatic microplate with respect to the Eurasian and African plates is responsible for the Mesozoic to present tectonic evolution of the Alps, Carpathians, the Dinarides and Hellenides as well as the Apennines. The classical approach for reconstructing plate motions is to assume that tectonic plates are rigid, then apply Euler's theorem to describe their rotation on an ideally spherical Earth by stepwise restorations of magnetic anomalies and fracture zones in oceanic basins. However, this approach is inadequate for reconstructing the motion of Mediterranean microplates like Adria, which, at present, is surrounded by convergent margins and whose oceanic portions have by now been entirely subducted. Most constraints on the motion of the Adriatic microplate come either from palaeomagnetics or from shortening estimates in the Alps, i.e., its northern margin. This approach renders plate tectonic reconstructions prone to numerous errors, yielding inadmissible misfits in the Ionian Sea between southern Italy and northern Greece. At the same time, Adria's western and eastern margins in the Apennines and in the Dinarides have hitherto not been appropriately considered for improving constraints on the motion of Adria. This presentation presents new results of ongoing collaborative research that aims at improving the relative motion path for the Adriatic microplate for the Cenozoic by additionally quantifying and restoring the amount of shortening and extension in a set of geophysical-geological transects from the Tyrrhenian Sea, the Apennines and the Dinarides. Already now, our approach yields an improved motion path for the Adriatic microplate for the last 20 Ma, which minimizes misfits in previous reconstructions. The currently largest challenge in our reconstructions is to reconcile amount and age of shortening in the Dinarides fold-and-thrust belt. For one thing, we see good agreement between the cross-sectional length of subducted material (c. 135 km

Genetically Encoded Biosensors in Plants: Pathways to Discovery.

PubMed

Walia, Ankit; Waadt, Rainer; Jones, Alexander M

2018-04-29

Genetically encoded biosensors that directly interact with a molecule of interest were first introduced more than 20 years ago with fusion proteins that served as fluorescent indicators for calcium ions. Since then, the technology has matured into a diverse array of biosensors that have been deployed to improve our spatiotemporal understanding of molecules whose dynamics have profound influence on plant physiology and development. In this review, we address several types of biosensors with a focus on genetically encoded calcium indicators, which are now the most diverse and advanced group of biosensors. We then consider the discoveries in plant biology made by using biosensors for calcium, pH, reactive oxygen species, redox conditions, primary metabolites, phytohormones, and nutrients. These discoveries were dependent on the engineering, characterization, and optimization required to develop a successful biosensor; they were also dependent on the methodological developments required to express, detect, and analyze the readout of such biosensors.

Yeast-based biosensors: design and applications.

PubMed

Adeniran, Adebola; Sherer, Michael; Tyo, Keith E J

2015-02-01

Yeast-based biosensing (YBB) is an exciting research area, as many studies have demonstrated the use of yeasts to accurately detect specific molecules. Biosensors incorporating various yeasts have been reported to detect an incredibly large range of molecules including but not limited to odorants, metals, intracellular metabolites, carcinogens, lactate, alcohols, and sugars. We review the detection strategies available for different types of analytes, as well as the wide range of output methods that have been incorporated with yeast biosensors. We group biosensors into two categories: those that are dependent upon transcription of a gene to report the detection of a desired molecule and those that are independent of this reporting mechanism. Transcription-dependent biosensors frequently depend on heterologous expression of sensing elements from non-yeast organisms, a strategy that has greatly expanded the range of molecules available for detection by YBBs. Transcription-independent biosensors circumvent the problem of sensing difficult-to-detect analytes by instead relying on yeast metabolism to generate easily detected molecules when the analyte is present. The use of yeast as the sensing element in biosensors has proven to be successful and continues to hold great promise for a variety of applications. © FEMS 2015. All rights reserved. For permissions, please e-mail: journals.permission@oup.com.

Recent Development in Optical Fiber Biosensors

PubMed Central

Bosch, María Espinosa; Sánchez, Antonio Jesús Ruiz; Rojas, Fuensanta Sánchez; Ojeda, Catalina Bosch

2007-01-01

Remarkable developments can be seen in the field of optical fibre biosensors in the last decade. More sensors for specific analytes have been reported, novel sensing chemistries or transduction principles have been introduced, and applications in various analytical fields have been realised. This review consists of papers mainly reported in the last decade and presents about applications of optical fiber biosensors. Discussions on the trends in optical fiber biosensor applications in real samples are enumerated.

Fiber optic-based biosensor

NASA Technical Reports Server (NTRS)

Ligler, Frances S.

1991-01-01

The NRL fiber optic biosensor is a device which measures the formation of a fluorescent complex at the surface of an optical fiber. Antibodies and DNA binding proteins provide the mechanism for recognizing an analyze and immobilizing a fluorescent complex on the fiber surface. The fiber optic biosensor is fast, sensitive, and permits analysis of hazardous materials remote from the instrumentation. The fiber optic biosensor is described in terms of the device configuration, chemistry for protein immobilization, and assay development. A lab version is being used for assay development and performance characterization while a portable device is under development. Antibodies coated on the fiber are stable for up to two years of storage prior to use. The fiber optic biosensor was used to measure concentration of toxins in the parts per billion (ng/ml) range in under a minute. Immunoassays for small molecules and whole bacteria are under development. Assays using DNA probes as the detection element can also be used with the fiber optic sensor, which is currently being developed to detect biological warfare agents, explosives, pathogens, and toxic materials which pollute the environment.

Single bead-based electrochemical biosensor.

PubMed

Liu, Changchun; Schrlau, Michael G; Bau, Haim H

2009-12-15

A simple, robust, single bead-based electrochemical biosensor was fabricated and characterized. The sensor's working electrode consists of an electrochemically etched platinum wire, with a nominal diameter of 25 microm, hermetically heat-fusion sealed in a pulled glass capillary (micropipette). The sealing process does not require any epoxy or glue. A commercially available, densely functionalized agarose bead was mounted on the tip of the etched platinum wire. The use of a pre-functionalized bead eliminates the tedious and complicated surface functionalization process that is often the bottleneck in the development of electrochemical biosensors. We report on the use of a biotin agarose bead-based, micropipette, electrochemical (Bio-BMP) biosensor to monitor H(2)O(2) concentration and the use of a streptavidin bead-based, micropipette, electrochemical (SA-BMP) biosensor to detect DNA amplicons. The Bio-BMP biosensor's response increased linearly as the H(2)O(2) concentration increased in the range from 1 x 10(-6) to 1.2 x10(-4)M with a detection limit of 5 x 10(-7)M. The SA-BMP was able to detect the amplicons of 1pg DNA template of B. Cereus bacteria, thus providing better detection sensitivity than conventional gel-based electropherograms.

A simple and rapid microplate assay for glycoprotein-processing glycosidases

DOE Office of Scientific and Technical Information (OSTI.GOV)

Kang, M.S.; Zwolshen, J.H.; Harry, B.S.

1989-08-15

A simple and convenient microplate assay for glycosidases involved in the glycoprotein-processing reactions is described. The assay is based on specific binding of high-mannose-type oligosaccharide substrates to concanavalin A-Sepharose, while monosaccharides liberated by enzymatic hydrolysis do not bind to concanavalin A-Sepharose. By the use of radiolabeled substrates (( 3H)glucose for glucosidases and (3H)mannose for mannosidases), the radioactivity in the liberated monosaccharides can be determined as a measure of the enzymatic activity. This principle was employed earlier for developing assays for glycosidases previously reported. These authors have reported the separation of substrate from the product by concanavalin A-Sepharose column chromatography. Thismore » procedure is handicapped by the fact that it cannot be used for a large number of samples and is time consuming. We have simplified this procedure and adapted it to the use of a microplate (96-well plate). This would help in processing a large number of samples in a short time. In this report we show that the assay is comparable to the column assay previously reported. It is linear with time and enzyme concentration and shows expected kinetics with castanospermine, a known inhibitor of alpha-glucosidase I.« less

Development of biosensors based on the one-dimensional semiconductor nanomaterials.

PubMed

Yan, Shancheng; Shi, Yi; Xiao, Zhongdang; Zhou, Minmin; Yan, Wenfu; Shen, Haoliang; Hu, Dong

2012-09-01

Biosensors are becoming increasingly important due to their applications in biological and chemical analyses, food safety industry, biomedical diagnostics, clinical detection, and environmental monitoring. Recent years, nanostructured semiconductor materials have been used to fabricate biosensors owing to their biocompatibility, low toxicity, high electron mobility, and easy fabrication. In the present study, we focus on recent various biosensors based on the one-dimensional semiconductor nanomaterials such as electrochemical biosensor, field-effect transistors biosensor, and label-free optical biosensor. In particular, the development of the electrochemical biosensor is discussed detailedly.

Synthetic biology for microbial heavy metal biosensors.

PubMed

Kim, Hyun Ju; Jeong, Haeyoung; Lee, Sang Jun

2018-02-01

Using recombinant DNA technology, various whole-cell biosensors have been developed for detection of environmental pollutants, including heavy metal ions. Whole-cell biosensors have several advantages: easy and inexpensive cultivation, multiple assays, and no requirement of any special techniques for analysis. In the era of synthetic biology, cutting-edge DNA sequencing and gene synthesis technologies have accelerated the development of cell-based biosensors. Here, we summarize current technological advances in whole-cell heavy metal biosensors, including the synthetic biological components (bioparts), sensing and reporter modules, genetic circuits, and chassis cells. We discuss several opportunities for improvement of synthetic cell-based biosensors. First, new functional modules must be discovered in genome databases, and this knowledge must be used to upgrade specific bioparts through molecular engineering. Second, modules must be assembled into functional biosystems in chassis cells. Third, heterogeneity of individual cells in the microbial population must be eliminated. In the perspectives, the development of whole-cell biosensors is also discussed in the aspects of cultivation methods and synthetic cells.

Biosensor Architectures for High-Fidelity Reporting of Cellular Signaling

PubMed Central

Dushek, Omer; Lellouch, Annemarie C.; Vaux, David J.; Shahrezaei, Vahid

2014-01-01

Understanding mechanisms of information processing in cellular signaling networks requires quantitative measurements of protein activities in living cells. Biosensors are molecular probes that have been developed to directly track the activity of specific signaling proteins and their use is revolutionizing our understanding of signal transduction. The use of biosensors relies on the assumption that their activity is linearly proportional to the activity of the signaling protein they have been engineered to track. We use mechanistic mathematical models of common biosensor architectures (single-chain FRET-based biosensors), which include both intramolecular and intermolecular reactions, to study the validity of the linearity assumption. As a result of the classic mechanism of zero-order ultrasensitivity, we find that biosensor activity can be highly nonlinear so that small changes in signaling protein activity can give rise to large changes in biosensor activity and vice versa. This nonlinearity is abolished in architectures that favor the formation of biosensor oligomers, but oligomeric biosensors produce complicated FRET states. Based on this finding, we show that high-fidelity reporting is possible when a single-chain intermolecular biosensor is used that cannot undergo intramolecular reactions and is restricted to forming dimers. We provide phase diagrams that compare various trade-offs, including observer effects, which further highlight the utility of biosensor architectures that favor intermolecular over intramolecular binding. We discuss challenges in calibrating and constructing biosensors and highlight the utility of mathematical models in designing novel probes for cellular signaling. PMID:25099816

Alps to Apennines zircon roller coaster along the Adria microplate margin.

PubMed

Jacobs, J; Paoli, G; Rocchi, S; Ksienzyk, A K; Sirevaag, H; Elburg, M A

2018-02-09

We have traced the particle path of high-pressure metasedimentary rocks on Elba Island, Northern Apennines, with the help of a U-Pb-Hf detrital zircon study. One quarter of the analysed zircons are surprisingly young, 41-30 Ma, with a main age peak at ca. 32 Ma, indicating an unexpected early Oligocene maximum deposition age. These Oligocene ages with negative εHf indicate a volcanic source region in the central-southern Alps. Though young by geological means, these zircons record an extraordinary geodynamic history. They originated in a volcanic arc, during the convergence/collision of the the Adria microplate with Europe from ca. 65 to 30 Ma. Thereafter, the Oligocene zircons travelled ca. 400 km southward along the Adria margin and the accretionary prism to present-day Tuscany, where they were subducted to depths of at least 40 km. Shortly thereafter, they were brought to the surface again in the wake of hinge roll back of the Apennine subduction zone and the resulting rapid extensional exhumation. Such a zircon roller coaster requires a microplate that has back-to-back subduction zones with opposing polarities on two sides.

BIOSENSORS

EPA Science Inventory

It has recently been proposed under the International Union of Pure and Applied Chemistry (IUPAC) Commission that biosensors be regarded as a subgroup of chemical sensors in which a biologically based mechanism is used for detection of the analyte. hemical sensors are defined und...

A creatinine biosensor based on admittance measurement

NASA Astrophysics Data System (ADS)

Ching, Congo Tak-Shing; Sun, Tai-Ping; Jheng, Deng-Yun; Tsai, Hou-Wei; Shieh, Hsiu-Li

2015-08-01

Regular check of blood creatinine level is very important as it is a measurement of renal function. Therefore, the objective of this study is to develop a simple and reliable creatinine biosensor based on admittance measurement for precise determination of creatinine. The creatinine biosensor was fabricated with creatinine deiminase immobilized on screen-printed carbon electrodes. Admittance measurement at a specific frequency ranges (22.80 - 84.71 Hz) showed that the biosensor has an excellent linear (r2 > 0.95) response range (50 - 250 uM), which covers the normal physiological and pathological ranges of blood creatinine levels. Intraclass correlation coefficient (ICC) showed that the biosensor has excellent reliability and validity (ICC = 0.98). In conclusion, a simple and reliable creatinine biosensor was developed and it is capable of precisely determining blood creatinine levels in both the normal physiological and pathological ranges.

Functionalized xenon as a biosensor

PubMed Central

Spence, Megan M.; Rubin, Seth M.; Dimitrov, Ivan E.; Ruiz, E. Janette; Wemmer, David E.; Pines, Alexander; Yao, Shao Qin; Tian, Feng; Schultz, Peter G.

2001-01-01

The detection of biological molecules and their interactions is a significant component of modern biomedical research. In current biosensor technologies, simultaneous detection is limited to a small number of analytes by the spectral overlap of their signals. We have developed an NMR-based xenon biosensor that capitalizes on the enhanced signal-to-noise, spectral simplicity, and chemical-shift sensitivity of laser-polarized xenon to detect specific biomolecules at the level of tens of nanomoles. We present results using xenon “functionalized” by a biotin-modified supramolecular cage to detect biotin–avidin binding. This biosensor methodology can be extended to a multiplexing assay for multiple analytes. PMID:11535830

Fabrication of single phase 2D homologous perovskite microplates by mechanical exfoliation

NASA Astrophysics Data System (ADS)

Li, Junze; Wang, Jun; Zhang, Yingjun; Wang, Haizhen; Lin, Gaoming; Xiong, Xuan; Zhou, Weihang; Luo, Hongmei; Li, Dehui

2018-04-01

The two-dimensional (2D) Ruddlesden-Popper type perovskites have attracted intensive interest for their great environmental stability and various potential optoelectronic applications. Fundamental understanding of the photophysical and electronic properties of the 2D perovskites with pure single phase is essential for improving the performance of the optoelectronic devices and designing devices with new architectures. Investigating the optical and electronic properties of these materials with pure single phase is required to obtain pure single phase 2D perovskites. Here, we report on an alternative approach to fabricate (C4H9NH3)2(CH3NH3) n-1Pb n I3n+1 microplates with pure single n-number perovskite phase for n  >  2 by mechanical exfoliation. Micro-photoluminescence and absorption spectroscopy studies reveal that the as-synthesized 2D perovskite plates for n  >  2 are comprised by dominant n-number phase and small inclusions of hybrid perovskite phases with different n values, which is supported by excitation power dependent photoluminescence. By mechanical exfoliation method, 2D perovskite microplates with the thickness of around 20 nm are obtained, which surprisingly have single n-number perovskite phase for n  =  2-5. In addition, we have demonstrated that the exfoliated 2D perovskite microplates can be integrated with other 2D layered materials such as boron nitride, and are able to be transferred to prefabricated electrodes for photodetections. Our studies not only provide a strategy to prepare 2D perovskites with a single n-number perovskite phase allowing us to extract the basic optical and electronic parameters of pure phase perovskites, but also demonstrate the possibility to integrate the 2D perovskites with other 2D layered materials to extend the device’s functionalities.

Anaerobic microplate assay for direct microbial conversion of switchgrass and Avicel using Clostridium thermocellum.

PubMed

Oguntimein, Gbekeloluwa B; Rodriguez, Miguel; Dumitrache, Alexandru; Shollenberger, Todd; Decker, Stephen R; Davison, Brian H; Brown, Steven D

2018-02-01

To develop and prototype a high-throughput microplate assay to assess anaerobic microorganisms and lignocellulosic biomasses in a rapid, cost-effective screen for consolidated bioprocessing potential. Clostridium thermocellum parent Δhpt strain deconstructed Avicel to cellobiose, glucose, and generated lactic acid, formic acid, acetic acid and ethanol as fermentation products in titers and ratios similar to larger scale fermentations confirming the suitability of a plate-based method for C. thermocellum growth studies. C. thermocellum strain LL1210, with gene deletions in the key central metabolic pathways, produced higher ethanol titers in the Consolidated Bioprocessing (CBP) plate assay for both Avicel and switchgrass fermentations when compared to the Δhpt strain. A prototype microplate assay system is developed that will facilitate high-throughput bioprospecting for new lignocellulosic biomass types, genetic variants and new microbial strains for bioethanol production.

Anaerobic microplate assay for direct microbial conversion of switchgrass and Avicel using Clostridium thermocellum

DOE PAGES

Oguntimein, Gbekeloluwa B.; Rodriguez, Jr., Miguel; Dumitrache, Alexandru; ...

2017-11-09

Here, to develop and prototype a high-throughput microplate assay to assess anaerobic microorganisms and lignocellulosic biomasses in a rapid, cost-effective screen for consolidated bioprocessing potential. Clostridium thermocellum parent Δ hpt strain deconstructed Avicel to cellobiose, glucose, and generated lactic acid, formic acid, acetic acid and ethanol as fermentation products in titers and ratios similar to larger scale fermentations confirming the suitability of a plate-based method for C. thermocellum growth studies. C. thermocellum strain LL1210, with gene deletions in the key central metabolic pathways, produced higher ethanol titers in the Consolidated Bioprocessing (CBP) plate assay for both Avicel and switchgrass fermentationsmore » when compared to the Δ hpt strain. A prototype microplate assay system is developed that will facilitate high-throughput bioprospecting for new lignocellulosic biomass types, genetic variants and new microbial strains for bioethanol production.« less

Anaerobic microplate assay for direct microbial conversion of switchgrass and Avicel using Clostridium thermocellum

DOE Office of Scientific and Technical Information (OSTI.GOV)

Oguntimein, Gbekeloluwa B.; Rodriguez, Jr., Miguel; Dumitrache, Alexandru

Here, to develop and prototype a high-throughput microplate assay to assess anaerobic microorganisms and lignocellulosic biomasses in a rapid, cost-effective screen for consolidated bioprocessing potential. Clostridium thermocellum parent Δ hpt strain deconstructed Avicel to cellobiose, glucose, and generated lactic acid, formic acid, acetic acid and ethanol as fermentation products in titers and ratios similar to larger scale fermentations confirming the suitability of a plate-based method for C. thermocellum growth studies. C. thermocellum strain LL1210, with gene deletions in the key central metabolic pathways, produced higher ethanol titers in the Consolidated Bioprocessing (CBP) plate assay for both Avicel and switchgrass fermentationsmore » when compared to the Δ hpt strain. A prototype microplate assay system is developed that will facilitate high-throughput bioprospecting for new lignocellulosic biomass types, genetic variants and new microbial strains for bioethanol production.« less

Molecular Rotors for Universal Quantitation of Nanoscale Hydrophobic Interfaces in Microplate Format.

PubMed

Bisso, Paul W; Tai, Michelle; Katepalli, Hari; Bertrand, Nicolas; Blankschtein, Daniel; Langer, Robert

2018-01-10

Hydrophobic self-assembly pairs diverse chemical precursors and simple formulation processes to access a vast array of functional colloids. Exploration of this design space, however, is stymied by lack of broadly general, high-throughput colloid characterization tools. Here, we show that a narrow structural subset of fluorescent, zwitterionic molecular rotors, dialkylaminostilbazolium sulfonates [DASS] with intermediate-length alkyl tails, fills this major analytical void by quantitatively sensing hydrophobic interfaces in microplate format. DASS dyes supersede existing interfacial probes by avoiding off-target fluorogenic interactions and dye aggregation while preserving hydrophobic partitioning strength. To illustrate the generality of this approach, we demonstrate (i) a microplate-based technique for measuring mass concentration of small (20-200 nm), dilute (submicrogram sensitivity) drug delivery nanoparticles; (ii) elimination of particle size, surfactant chemistry, and throughput constraints on quantifying the complex surfactant/metal oxide adsorption isotherms critical for environmental remediation and enhanced oil recovery; and (iii) more reliable self-assembly onset quantitation for chemically and structurally distinct amphiphiles. These methods could streamline the development of nanotechnologies for a broad range of applications.

NANOSCALE BIOSENSORS IN ECOSYSTEM EXPOSURE RESEARCH

EPA Science Inventory

This powerpoint presentation presented information on nanoscale biosensors in ecosystem exposure research. The outline of the presentation is as follows: nanomaterials environmental exposure research; US agencies involved in nanosensor research; nanoscale LEDs in biosensors; nano...

Peptide code-on-a-microplate for protease activity analysis via MALDI-TOF mass spectrometric quantitation.

PubMed

Hu, Junjie; Liu, Fei; Ju, Huangxian

2015-04-21

A peptide-encoded microplate was proposed for MALDI-TOF mass spectrometric (MS) analysis of protease activity. The peptide codes were designed to contain a coding region and the substrate of protease for enzymatic cleavage, respectively, and an internal standard method was proposed for the MS quantitation of the cleavage products of these peptide codes. Upon the cleavage reaction in the presence of target proteases, the coding regions were released from the microplate, which were directly quantitated by using corresponding peptides with one-amino acid difference as the internal standards. The coding region could be used as the unique "Protease ID" for the identification of corresponding protease, and the amount of the cleavage product was used for protease activity analysis. Using trypsin and chymotrypsin as the model proteases to verify the multiplex protease assay, the designed "Trypsin ID" and "Chymotrypsin ID" occurred at m/z 761.6 and 711.6. The logarithm value of the intensity ratio of "Protease ID" to internal standard was proportional to trypsin and chymotrypsin concentration in a range from 5.0 to 500 and 10 to 500 nM, respectively. The detection limits for trypsin and chymotrypsin were 2.3 and 5.2 nM, respectively. The peptide-encoded microplate showed good selectivity. This proposed method provided a powerful tool for convenient identification and activity analysis of multiplex proteases.

Affinity Biosensors for Detection of Mycotoxins in Food.

PubMed

Evtugyn, Gennady; Subjakova, Veronika; Melikishvili, Sopio; Hianik, Tibor

2018-01-01

This chapter reviews recent achievements in methods of detection of mycotoxins in food. Special focus is on the biosensor technology that utilizes antibodies and nucleic acid aptamers as receptors. Development of biosensors is based on the immobilization of antibodies or aptamers onto various conventional supports like gold layer, but also on nanomaterials such as graphene oxide, carbon nanotubes, and quantum dots that provide an effective platform for achieving high sensitivity of detection using various physical methods, including electrochemical, mass sensitive, and optical. The biosensors developed so far demonstrate high sensitivity typically in subnanomolar limit of detection. Several biosensors have been validated in real samples. The sensitivity of biosensors is similar and, in some cases, even better than traditional analytical methods such as ELISA or chromatography. We believe that future trends will be focused on improving biosensor properties toward practical application in food industry. © 2018 Elsevier Inc. All rights reserved.

Microplate-based platform for combined chromatin and DNA methylation immunoprecipitation assays

PubMed Central

2011-01-01

Background The processes that compose expression of a given gene are far more complex than previously thought presenting unprecedented conceptual and mechanistic challenges that require development of new tools. Chromatin structure, which is regulated by DNA methylation and histone modification, is at the center of gene regulation. Immunoprecipitations of chromatin (ChIP) and methylated DNA (MeDIP) represent a major achievement in this area that allow researchers to probe chromatin modifications as well as specific protein-DNA interactions in vivo and to estimate the density of proteins at specific sites genome-wide. Although a critical component of chromatin structure, DNA methylation has often been studied independently of other chromatin events and transcription. Results To allow simultaneous measurements of DNA methylation with other genomic processes, we developed and validated a simple and easy-to-use high throughput microplate-based platform for analysis of DNA methylation. Compared to the traditional beads-based MeDIP the microplate MeDIP was more sensitive and had lower non-specific binding. We integrated the MeDIP method with a microplate ChIP assay which allows measurements of both DNA methylation and histone marks at the same time, Matrix ChIP-MeDIP platform. We illustrated several applications of this platform to relate DNA methylation, with chromatin and transcription events at selected genes in cultured cells, human cancer and in a model of diabetic kidney disease. Conclusion The high throughput capacity of Matrix ChIP-MeDIP to profile tens and potentially hundreds of different genomic events at the same time as DNA methylation represents a powerful platform to explore complex genomic mechanism at selected genes in cultured cells and in whole tissues. In this regard, Matrix ChIP-MeDIP should be useful to complement genome-wide studies where the rich chromatin and transcription database resources provide fruitful foundation to pursue mechanistic

Protein Detection with Aptamer Biosensors

PubMed Central

Strehlitz, Beate; Nikolaus, Nadia; Stoltenburg, Regina

2008-01-01

Aptamers have been developed for different applications. Their use as new biological recognition elements in biosensors promises progress for fast and easy detection of proteins. This new generation of biosensor (aptasensors) will be more stable and well adapted to the conditions of real samples because of the specific properties of aptamers. PMID:27879936

Amperometric biosensor for Salmonella typhimurium detection in milk

USDA-ARS?s Scientific Manuscript database

This paper reports an amperometric biosensor for rapid and sensitive Salmonella Typhimurium detection in milk. The biosensor was assembled from the self-assembled monolayers technique on a gold surface. In this device, polyclonal antibodies were oriented by protein A. The biosensor structure was cha...

In vitro evaluation of fluorescence glucose biosensor response.

PubMed

Aloraefy, Mamdouh; Pfefer, T Joshua; Ramella-Roman, Jessica C; Sapsford, Kim E

2014-07-08

Rapid, accurate, and minimally-invasive glucose biosensors based on Förster Resonance Energy Transfer (FRET) for glucose measurement have the potential to enhance diabetes control. However, a standard set of in vitro approaches for evaluating optical glucose biosensor response under controlled conditions would facilitate technological innovation and clinical translation. Towards this end, we have identified key characteristics and response test methods, fabricated FRET-based glucose biosensors, and characterized biosensor performance using these test methods. The biosensors were based on competitive binding between dextran and glucose to concanavalin A and incorporated long-wavelength fluorescence dye pairs. Testing characteristics included spectral response, linearity, sensitivity, limit of detection, kinetic response, reversibility, stability, precision, and accuracy. The biosensor demonstrated a fluorescence change of 45% in the presence of 400 mg/dL glucose, a mean absolute relative difference of less than 11%, a limit of detection of 25 mg/dL, a response time of 15 min, and a decay in fluorescence intensity of 72% over 30 days. The battery of tests presented here for objective, quantitative in vitro evaluation of FRET glucose biosensors performance have the potential to form the basis of future consensus standards. By implementing these test methods for a long-visible-wavelength biosensor, we were able to demonstrate strengths and weaknesses with a new level of thoroughness and rigor.

In Vitro Evaluation of Fluorescence Glucose Biosensor Response

PubMed Central

Aloraefy, Mamdouh; Pfefer, T. Joshua; Ramella-Roman, Jessica C.; Sapsford, Kim E.

2014-01-01

Rapid, accurate, and minimally-invasive glucose biosensors based on Förster Resonance Energy Transfer (FRET) for glucose measurement have the potential to enhance diabetes control. However, a standard set of in vitro approaches for evaluating optical glucose biosensor response under controlled conditions would facilitate technological innovation and clinical translation. Towards this end, we have identified key characteristics and response test methods, fabricated FRET-based glucose biosensors, and characterized biosensor performance using these test methods. The biosensors were based on competitive binding between dextran and glucose to concanavalin A and incorporated long-wavelength fluorescence dye pairs. Testing characteristics included spectral response, linearity, sensitivity, limit of detection, kinetic response, reversibility, stability, precision, and accuracy. The biosensor demonstrated a fluorescence change of 45% in the presence of 400 mg/dL glucose, a mean absolute relative difference of less than 11%, a limit of detection of 25 mg/dL, a response time of 15 min, and a decay in fluorescence intensity of 72% over 30 days. The battery of tests presented here for objective, quantitative in vitro evaluation of FRET glucose biosensors performance have the potential to form the basis of future consensus standards. By implementing these test methods for a long-visible-wavelength biosensor, we were able to demonstrate strengths and weaknesses with a new level of thoroughness and rigor. PMID:25006996

Development, optimization and validation of a rapid colorimetric microplate bioassay for neomycin sulfate in pharmaceutical drug products.

PubMed

Francisco, Fabiane Lacerda; Saviano, Alessandro Morais; Pinto, Terezinha de Jesus Andreoli; Lourenço, Felipe Rebello

2014-08-01

Microbiological assays have been used to evaluate antimicrobial activity since the discovery of the first antibiotics. Despite their limitations, microbiological assays are widely employed to determine antibiotic potency of pharmaceutical dosage forms, since they provide a measure of biological activity. The aim of this work is to develop, optimize and validate a rapid colorimetric microplate bioassay for the potency of neomycin in pharmaceutical drug products. Factorial and response surface methodologies were used in the development and optimization of the choice of microorganism, culture medium composition, amount of inoculum, triphenyltetrazolium chloride (TTC) concentration and neomycin concentration. The optimized bioassay method was validated by the assessment of linearity (range 3.0 to 5.0μg/mL, r=0.998 and 0.994 for standard and sample curves, respectively), precision (relative standard deviation (RSD) of 2.8% and 4.0 for repeatability and intermediate precision, respectively), accuracy (mean recovery=100.2%) and robustness. Statistical analysis showed equivalency between agar diffusion microbiological assay and rapid colorimetric microplate bioassay. In addition, microplate bioassay had advantages concerning the sensitivity of response, time of incubation, and amount of culture medium and solutions required. Copyright © 2014 Elsevier B.V. All rights reserved.

The role of high-pressure metamorphic rocks in collisional processes with a microplate involved

NASA Astrophysics Data System (ADS)

Massonne, Hans-Joachim

2013-04-01

High-pressure (HP: p >10 kbar) rocks, especially easily recognizable eclogite lenses, are markers of a collision of major continental plates in Phanerozoic times, but how about a collison with a microplate? To answer this question, petrological studies were undertaken on garnet-bearing metamorphic rocks. Especially pressure (P) - temperature (T) pseudosections were calculated to reconstruct the P-T path of such rocks. Areas in the Andes and their eastern foreland (Argentina, Ecuador) and the Carboniferous realm of western and central Europe were studied, where microplates either had collided with the South American plate or had been located between colliding Laurussia and Gondwana. The following features resulted: (1) laterally extended zones of HP rocks mark collisional sutures, but eclogite lenses are virtually absent; (2) these HP rocks are dominantly metapelites which have experienced pressures as high as 1.2 to 1.5 GPa; (3) these pressures were reached between 450 to 550°C, followed by a P release at increasing T; (4) the T peak in the range of 500-650°C at P between 0.5 and 0.8 GPa is often characterized by the blastesis of plagioclase occasionally together with staurolite; (5) this event was pre-dated by two deformational events. The second deformation occurred during the early exhumation. Considering the regional geology of the study areas, these features are interpreted as follows: Prior to the collision a basin, extending over at most some hundred kilometers perpendicular to the basin axis, existed between the colliding microplate and the major plate. Eventually, a rifting event had caused the separation of the microplate from the major continental plate. The basin, which was opened by this event and filled with sediments, could have developed further to a back-arc basin. Then, the extensional regime turned to a compressional one. During compression the basin sediments on top of thinned continental crust were buried beneath the overriding major plate

Novel amperometric glucose biosensor based on MXene nanocomposite.

PubMed

Rakhi, R B; Nayak, Pranati; Xia, Chuan; Alshareef, Husam N

2016-11-10

A biosensor platform based on Au/MXene nanocomposite for sensitive enzymatic glucose detection is reported. The biosensor leverages the unique electrocatalytic properties and synergistic effects between Au nanoparticles and MXene sheets. An amperometric glucose biosensor is fabricated by the immobilization of glucose oxidase (GOx) enzyme on Nafion solubilized Au/ MXene nanocomposite over glassy carbon electrode (GCE). The biomediated Au nanoparticles play a significant role in facilitating the electron exchange between the electroactive center of GOx and the electrode. The GOx/Au/MXene/Nafion/GCE biosensor electrode displayed a linear amperometric response in the glucose concentration range from 0.1 to 18 mM with a relatively high sensitivity of 4.2 μAmM -1 cm -2 and a detection limit of 5.9 μM (S/N = 3). Furthermore, the biosensor exhibited excellent stability, reproducibility and repeatability. Therefore, the Au/MXene nanocomposite reported in this work is a potential candidate as an electrochemical transducer in electrochemical biosensors.

Novel amperometric glucose biosensor based on MXene nanocomposite

PubMed Central

Rakhi, R. B.; Nayuk, Pranati; Xia, Chuan; Alshareef, Husam N.

2016-01-01

A biosensor platform based on Au/MXene nanocomposite for sensitive enzymatic glucose detection is reported. The biosensor leverages the unique electrocatalytic properties and synergistic effects between Au nanoparticles and MXene sheets. An amperometric glucose biosensor is fabricated by the immobilization of glucose oxidase (GOx) enzyme on Nafion solubilized Au/ MXene nanocomposite over glassy carbon electrode (GCE). The biomediated Au nanoparticles play a significant role in facilitating the electron exchange between the electroactive center of GOx and the electrode. The GOx/Au/MXene/Nafion/GCE biosensor electrode displayed a linear amperometric response in the glucose concentration range from 0.1 to 18 mM with a relatively high sensitivity of 4.2 μAmM−1 cm−2 and a detection limit of 5.9 μM (S/N = 3). Furthermore, the biosensor exhibited excellent stability, reproducibility and repeatability. Therefore, the Au/MXene nanocomposite reported in this work is a potential candidate as an electrochemical transducer in electrochemical biosensors. PMID:27830757

A Highly Responsive Silicon Nanowire/Amplifier MOSFET Hybrid Biosensor.

PubMed

Lee, Jieun; Jang, Jaeman; Choi, Bongsik; Yoon, Jinsu; Kim, Jee-Yeon; Choi, Yang-Kyu; Kim, Dong Myong; Kim, Dae Hwan; Choi, Sung-Jin

2015-07-21

This study demonstrates a hybrid biosensor comprised of a silicon nanowire (SiNW) integrated with an amplifier MOSFET to improve the current response of field-effect-transistor (FET)-based biosensors. The hybrid biosensor is fabricated using conventional CMOS technology, which has the potential advantage of high density and low noise performance. The biosensor shows a current response of 5.74 decades per pH for pH detection, which is 2.5 × 10(5) times larger than that of a single SiNW sensor. In addition, we demonstrate charged polymer detection using the biosensor, with a high current change of 4.5 × 10(5) with a 500 nM concentration of poly(allylamine hydrochloride). In addition, we demonstrate a wide dynamic range can be obtained by adjusting the liquid gate voltage. We expect that this biosensor will be advantageous and practical for biosensor applications which requires lower noise, high speed, and high density.

A Highly Responsive Silicon Nanowire/Amplifier MOSFET Hybrid Biosensor

PubMed Central

Lee, Jieun; Jang, Jaeman; Choi, Bongsik; Yoon, Jinsu; Kim, Jee-Yeon; Choi, Yang-Kyu; Myong Kim, Dong; Hwan Kim, Dae; Choi, Sung-Jin

2015-01-01

This study demonstrates a hybrid biosensor comprised of a silicon nanowire (SiNW) integrated with an amplifier MOSFET to improve the current response of field-effect-transistor (FET)-based biosensors. The hybrid biosensor is fabricated using conventional CMOS technology, which has the potential advantage of high density and low noise performance. The biosensor shows a current response of 5.74 decades per pH for pH detection, which is 2.5 × 105 times larger than that of a single SiNW sensor. In addition, we demonstrate charged polymer detection using the biosensor, with a high current change of 4.5 × 105 with a 500 nM concentration of poly(allylamine hydrochloride). In addition, we demonstrate a wide dynamic range can be obtained by adjusting the liquid gate voltage. We expect that this biosensor will be advantageous and practical for biosensor applications which requires lower noise, high speed, and high density. PMID:26197105

Disease-Related Detection with Electrochemical Biosensors: A Review.

PubMed

Huang, Ying; Xu, Jin; Liu, Junjie; Wang, Xiangyang; Chen, Bin

2017-10-17

Rapid diagnosis of diseases at their initial stage is critical for effective clinical outcomes and promotes general public health. Classical in vitro diagnostics require centralized laboratories, tedious work and large, expensive devices. In recent years, numerous electrochemical biosensors have been developed and proposed for detection of various diseases based on specific biomarkers taking advantage of their features, including sensitivity, selectivity, low cost and rapid response. This article reviews research trends in disease-related detection with electrochemical biosensors. Focus has been placed on the immobilization mechanism of electrochemical biosensors, and the techniques and materials used for the fabrication of biosensors are introduced in details. Various biomolecules used for different diseases have been listed. Besides, the advances and challenges of using electrochemical biosensors for disease-related applications are discussed.

Magnetically-refreshable receptor platform structures for reusable nano-biosensor chips

NASA Astrophysics Data System (ADS)

Yoo, Haneul; Lee, Dong Jun; Cho, Dong-guk; Park, Juhun; Nam, Ki Wan; Tak Cho, Young; Park, Jae Yeol; Chen, Xing; Hong, Seunghun

2016-01-01

We developed a magnetically-refreshable receptor platform structure which can be integrated with quite versatile nano-biosensor structures to build reusable nano-biosensor chips. This structure allows one to easily remove used receptor molecules from a biosensor surface and reuse the biosensor for repeated sensing operations. Using this structure, we demonstrated reusable immunofluorescence biosensors. Significantly, since our method allows one to place receptor molecules very close to a nano-biosensor surface, it can be utilized to build reusable carbon nanotube transistor-based biosensors which require receptor molecules within a Debye length from the sensor surface. Furthermore, we also show that a single sensor chip can be utilized to detect two different target molecules simply by replacing receptor molecules using our method. Since this method does not rely on any chemical reaction to refresh sensor chips, it can be utilized for versatile biosensor structures and virtually-general receptor molecular species.

Recent Advances in Nanotechnology Applied to Biosensors

PubMed Central

Zhang, Xueqing; Guo, Qin; Cui, Daxiang

2009-01-01

In recent years there has been great progress the application of nanomaterials in biosensors. The importance of these to the fundamental development of biosensors has been recognized. In particular, nanomaterials such as gold nanoparticles, carbon nanotubes, magnetic nanoparticles and quantum dots have been being actively investigated for their applications in biosensors, which have become a new interdisciplinary frontier between biological detection and material science. Here we review some of the main advances in this field over the past few years, explore the application prospects, and discuss the issues, approaches, and challenges, with the aim of stimulating a broader interest in developing nanomaterial-based biosensors and improving their applications in disease diagnosis and food safety examination. PMID:22399954

REVIEW ARTICLE: Environmental applications of analytical biosensors

NASA Astrophysics Data System (ADS)

Marco, María-Pilar; Barceló, Damià

1996-11-01

A review of the fundamental aspects and environmental applications of biosensors is presented. The bases of different transducer principles such as electrochemical, optical and piezoelectric are discussed. Various examples are given of the applications of such principles to develop immunosensor devices to determine common environmental contaminants. Attention is also paid to catalytic biosensors, using enzymes as sensing elements. Biosensor devices based on the use of cholinesterase and various oxidase enzymes such as tyrosinase, laccase, peroxidase and aldehyde dehydrogenase are reported. Some examples are given of the applications of other biomolecules such as whole cells, DNA or proteins, to determine pollution. Validation studies are presented comparing biosensors with chromatographic techniques to determine organophosphorus pesticides and phenolic compounds in environmental samples.

Biosensor technology for pesticides--a review.

PubMed

Verma, Neelam; Bhardwaj, Atul

2015-03-01

Pesticides, due to their lucrative outcomes, are majorly implicated in agricultural fields for crop production enhancement. Due to their pest removal properties, pesticides of various classes have been designed to persist in the environment over a longer duration after their application to achieve maximum effectiveness. Apart from their recalcitrant structure and agricultural benefits, pesticides also impose acute toxicological effects onto the other various life forms. Their accumulation in the living system may prove to be detrimental if established in higher concentrations. Thus, their prompt and accurate analysis is a crucial matter of concern. Conventional techniques like chromatographic techniques (HPLC, GC, etc.) used for pesticides detection are associated with various limitations like stumpy sensitivity and efficiency, time consumption, laboriousity, requirement of expensive equipments and highly trained technicians, and many more. So there is a need to recruit the methods which can detect these neurotoxic compounds sensitively, selectively, rapidly, and easily in the field. Present work is a brief review of the pesticide effects, their current usage scenario, permissible limits in various food stuffs and 21st century advancements of biosensor technology for pesticide detection. Due to their exceptional performance capabilities, easiness in operation and on-site working, numerous biosensors have been developed for bio-monitoring of various environmental samples for pesticide evaluation immensely throughout the globe. Till date, based on sensing element (enzyme based, antibody based, etc.) and type of detection method used (Electrochemical, optical, and piezoelectric, etc.), a number of biosensors have been developed for pesticide detection. In present communication, authors have summarized 21st century's approaches of biosensor technology for pesticide detection such as enzyme-based biosensors, immunosensors, aptamers, molecularly imprinted polymers, and

Micro-Photoluminescence (micro-PL) Study of Core-Shell GaAs/GaAsSb Nanowires Grown by Self-Assisted Molecular Beam Epitaxy

DTIC Science & Technology

2015-06-18

public release; distribution is unlimited. Micro-Photoluminescence (micro-PL) Study of Core-Shell GaAs/GaAsSb Nanowires grown by Self-Assisted Molecular...U.S. Army Research Office P.O. Box 12211 Research Triangle Park, NC 27709-2211 GaAsSb, Core Shell Nanowires , Micro Photoluminescence...University 1601 East Market Street Greensboro, NC 27411 -0001 ABSTRACT Micro-Photoluminescence (micro-PL) Study of Core-Shell GaAs/GaAsSb Nanowires grown by

Biosensor technology: technology push versus market pull.

PubMed

Luong, John H T; Male, Keith B; Glennon, Jeremy D

2008-01-01

Biosensor technology is based on a specific biological recognition element in combination with a transducer for signal processing. Since its inception, biosensors have been expected to play a significant analytical role in medicine, agriculture, food safety, homeland security, environmental and industrial monitoring. However, the commercialization of biosensor technology has significantly lagged behind the research output as reflected by a plethora of publications and patenting activities. The rationale behind the slow and limited technology transfer could be attributed to cost considerations and some key technical barriers. Analytical chemistry has changed considerably, driven by automation, miniaturization, and system integration with high throughput for multiple tasks. Such requirements pose a great challenge in biosensor technology which is often designed to detect one single or a few target analytes. Successful biosensors must be versatile to support interchangeable biorecognition elements, and in addition miniaturization must be feasible to allow automation for parallel sensing with ease of operation at a competitive cost. A significant upfront investment in research and development is a prerequisite in the commercialization of biosensors. The progress in such endeavors is incremental with limited success, thus, the market entry for a new venture is very difficult unless a niche product can be developed with a considerable market volume.

Characterization of growth inhibition of oral bacteria by sophorolipid using a microplate-format assay

USDA-ARS?s Scientific Manuscript database

Sophorolipid (SL) is a class of glycolipid biosurfactant produced by yeast and has potent antimicrobial activity against many microorganisms. In this paper, a microplate-based method was developed to characterize the growth inhibition by SL on five representative species of caries-causing oral bact...

Disease-Related Detection with Electrochemical Biosensors: A Review

PubMed Central

Huang, Ying; Xu, Jin; Liu, Junjie; Wang, Xiangyang; Chen, Bin

2017-01-01

Rapid diagnosis of diseases at their initial stage is critical for effective clinical outcomes and promotes general public health. Classical in vitro diagnostics require centralized laboratories, tedious work and large, expensive devices. In recent years, numerous electrochemical biosensors have been developed and proposed for detection of various diseases based on specific biomarkers taking advantage of their features, including sensitivity, selectivity, low cost and rapid response. This article reviews research trends in disease-related detection with electrochemical biosensors. Focus has been placed on the immobilization mechanism of electrochemical biosensors, and the techniques and materials used for the fabrication of biosensors are introduced in details. Various biomolecules used for different diseases have been listed. Besides, the advances and challenges of using electrochemical biosensors for disease-related applications are discussed. PMID:29039742

Advances in nano-scaled biosensors for biomedical applications.

PubMed

Wang, Jianling; Chen, Guihua; Jiang, Hui; Li, Zhiyong; Wang, Xuemei

2013-08-21

Recently, a growing amount of attention has been focused on the utility of biosensors for biomedical applications. Combined with nanomaterials and nanostructures, nano-scaled biosensors are installed for biomedical applications, such as pathogenic bacteria monitoring, virus recognition, disease biomarker detection, among others. These nano-biosensors offer a number of advantages and in many respects are ideally suited to biomedical applications, which could be made as extremely flexible devices, allowing biomedical analysis with speediness, excellent selectivity and high sensitivity. This minireview discusses the literature published in the latest years on the advances in biomedical applications of nano-scaled biosensors for disease bio-marking and detection, especially in bio-imaging and the diagnosis of pathological cells and viruses, monitoring pathogenic bacteria, thus providing insight into the future prospects of biosensors in relevant clinical applications.

Prototype amperometric biosensor for sialic acid determination.

PubMed

Marzouk, Sayed A M; Ashraf, S S; Tayyari, Khawla A Al

2007-02-15

This paper describes the first report on the development, characterization, and applications of a prototype amperometric biosensor for free sialic acid (SA). The sensor was constructed by the coimmobilization of two enzymes, i.e., N-acetylneuraminic acid aldolase and pyruvate oxidase, on a polyester microporous membrane, which was then mounted on top of a platinum disk electrode. The SA biosensor operation was based on the sequential action of the two enzymes to ultimately produce hydrogen peroxide, which was then detected by anodic amperometry at the platinum electrode. The surface of the platinum electrode was coated with an electropolymeric layer to enhance the biosensor selectivity in the presence of interfering oxidizable species. Optimization of the enzyme layer composition resulted in a fast and steady current response in phosphate buffer pH 7.2 at 37 degrees C. The limit of detection was 10 microM, and the response was linear to 3.5 mM (r = 0.9987). The prepared SA biosensors retained approximately 85% of their initial sensitivity after 8 days and showed excellent response reproducibility (CV = 2.3%). Utilization of a third enzyme, sialidase, expanded the scope of the present SA biosensor to determine bound sialic acid as well. The merits of the described biosensor allowed its successful application in determining SA in biological and pharmaceutical samples. The obtained results indicated that the presented SA biosensor should be a useful bioanalytical tool in several biological and clinical applications such as screening of SA as a nonspecific tumor marker as well as monitoring of tumor therapy.

Sense and sensitivity in bioprocessing-detecting cellular metabolites with biosensors.

PubMed

Dekker, Linda; Polizzi, Karen M

2017-10-01

Biosensors use biological elements to detect or quantify an analyte of interest. In bioprocessing, biosensors are employed to monitor key metabolites. There are two main types: fully biological systems or biological recognition coupled with physical/chemical detection. New developments in chemical biosensors include multiplexed detection using microfluidics. Synthetic biology can be used to engineer new biological biosensors with improved characteristics. Although there have been few biosensors developed for bioprocessing thus far, emerging trends can be applied in the future. A range of new platform technologies will enable rapid engineering of new biosensors based on transcriptional activation, riboswitches, and Förster Resonance Energy Transfer. However, translation to industry remains a challenge and more research into the robustness biosensors at scale is needed. Copyright © 2017 Elsevier Ltd. All rights reserved.

Fluidics cube for biosensor miniaturization

NASA Technical Reports Server (NTRS)

Dodson, J. M.; Feldstein, M. J.; Leatzow, D. M.; Flack, L. K.; Golden, J. P.; Ligler, F. S.

2001-01-01

To create a small, portable, fully automated biosensor, a compact means of fluid handling is required. We designed, manufactured, and tested a "fluidics cube" for such a purpose. This cube, made of thermoplastic, contains reservoirs and channels for liquid samples and reagents and operates without the use of any internal valves or meters; it is a passive fluid circuit that relies on pressure relief vents to control fluid movement. We demonstrate the ability of pressure relief vents to control fluid movement and show how to simply manufacture or modify the cube. Combined with the planar array biosensor developed at the Naval Research Laboratory, it brings us one step closer to realizing our goal of a handheld biosensor capable of analyzing multiple samples for multiple analytes.

Highly Multiplexed RNA Aptamer Selection using a Microplate-based Microcolumn Device.

PubMed

Reinholt, Sarah J; Ozer, Abdullah; Lis, John T; Craighead, Harold G

2016-07-19

We describe a multiplexed RNA aptamer selection to 19 different targets simultaneously using a microcolumn-based device, MEDUSA (Microplate-based Enrichment Device Used for the Selection of Aptamers), as well as a modified selection process, that significantly reduce the time and reagents needed for selections. We exploited MEDUSA's reconfigurable design between parallel and serially-connected microcolumns to enable the use of just 2 aliquots of starting library, and its 96-well microplate compatibility to enable the continued use of high-throughput techniques in downstream processes. Our modified selection protocol allowed us to perform the equivalent of a 10-cycle selection in the time it takes for 4 traditional selection cycles. Several aptamers were discovered with nanomolar dissociation constants. Furthermore, aptamers were identified that not only bound with high affinity, but also acted as inhibitors to significantly reduce the activity of their target protein, mouse decapping exoribonuclease (DXO). The aptamers resisted DXO's exoribonuclease activity, and in studies monitoring DXO's degradation of a 30-nucleotide substrate, less than 1 μM of aptamer demonstrated significant inhibition of DXO activity. This aptamer selection method using MEDUSA helps to overcome some of the major challenges with traditional aptamer selections, and provides a platform for high-throughput selections that lends itself to process automation.

Early Lung Cancer Diagnosis by Biosensors

PubMed Central

Zhang, Yuqian; Yang, Dongliang; Weng, Lixing; Wang, Lianhui

2013-01-01

Lung cancer causes an extreme threat to human health, and the mortality rate due to lung cancer has not decreased during the last decade. Prognosis or early diagnosis could help reduce the mortality rate. If microRNA and tumor-associated antigens (TAAs), as well as the corresponding autoantibodies, can be detected prior to clinical diagnosis, such high sensitivity of biosensors makes the early diagnosis and prognosis of cancer realizable. This review provides an overview of tumor-associated biomarker identifying methods and the biosensor technology available today. Laboratorial researches utilizing biosensors for early lung cancer diagnosis will be highlighted. PMID:23892596

A potentiometric biosensor for the detection of notch 3 using functionalized ZnO nanorods.

PubMed

Ibupoto, Z H; Khun, K; Liu, X; Willander, M

2014-09-01

The notch signalling plays a vital and radical role for the activity of cellular proliferation, differentiation and apoptosis. In this study, for the first time a particular biosensor is developed for the detection of notch 3. ZnO nanorods were fabricated on the gold coated glass substrate by hydrothermal method and afterwards were decorated with the gold nanoparticles by electrodepositing technique. Scanning electron microscopy (SEM) has shown the perpendicular to the substrate growth pattern of ZnO nanorods. X-ray diffraction (XRD) studies showed the c-axis oriented growth direction with wurtzite crystal structure of ZnO nanorods. X-ray Photoelectron Spectroscopy (XPS) and energy dispersive X-ray (EDX) techniques have shown the presence of Zn, O and Au atoms in the prepared functional material. Furthermore, the anti-notch 3 was physically adsorbed on the gold nanoparticles functionalized ZnO nanorods. The developed potentiometric immunosensor has shown response to the wide range of notch 3 molecules. The detected range included 1.00 x 10(-5)-1.50 x 10(0 ) μg/mL with a sensitivity of 23.15 ± 0.31 mV/decade. The analytical parameters including reproducibility, stability, and selectivity were also investigated and the observed results indicate the acceptable performance of the notch 3 biosensor. Moreover, the proposed notch 3 biosensor exhibited a fast response time of 10 s.

Solvent minimization induces preferential orientation and crystal clustering in serial micro-crystallography on micro-meshes, in situ plates and on a movable crystal conveyor belt.

PubMed

Soares, Alexei S; Mullen, Jeffrey D; Parekh, Ruchi M; McCarthy, Grace S; Roessler, Christian G; Jackimowicz, Rick; Skinner, John M; Orville, Allen M; Allaire, Marc; Sweet, Robert M

2014-11-01

X-ray diffraction data were obtained at the National Synchrotron Light Source from insulin and lysozyme crystals that were densely deposited on three types of surfaces suitable for serial micro-crystallography: MiTeGen MicroMeshes™, Greiner Bio-One Ltd in situ micro-plates, and a moving kapton crystal conveyor belt that is used to deliver crystals directly into the X-ray beam. 6° wedges of data were taken from ∼100 crystals mounted on each material, and these individual data sets were merged to form nine complete data sets (six from insulin crystals and three from lysozyme crystals). Insulin crystals have a parallelepiped habit with an extended flat face that preferentially aligned with the mounting surfaces, impacting the data collection strategy and the design of the serial crystallography apparatus. Lysozyme crystals had a cuboidal habit and showed no preferential orientation. Preferential orientation occluded regions of reciprocal space when the X-ray beam was incident normal to the data-collection medium surface, requiring a second pass of data collection with the apparatus inclined away from the orthogonal. In addition, crystals measuring less than 20 µm were observed to clump together into clusters of crystals. Clustering required that the X-ray beam be adjusted to match the crystal size to prevent overlapping diffraction patterns. No additional problems were encountered with the serial crystallography strategy of combining small randomly oriented wedges of data from a large number of specimens. High-quality data able to support a realistic molecular replacement solution were readily obtained from both crystal types using all three serial crystallography strategies.

Seismic evidence for rotating mantle flow around subducting slab edge associated with oceanic microplate capture

NASA Astrophysics Data System (ADS)

Mosher, Stephen G.; Audet, Pascal; L'Heureux, Ivan

2014-07-01

Tectonic plate reorganization at a subduction zone edge is a fundamental process that controls oceanic plate fragmentation and capture. However, the various factors responsible for these processes remain elusive. We characterize seismic anisotropy of the upper mantle in the Explorer region at the northern limit of the Cascadia subduction zone from teleseismic shear wave splitting measurements. Our results show that the mantle flow field beneath the Explorer slab is rotating anticlockwise from the convergence-parallel motion between the Juan de Fuca and the North America plates, re-aligning itself with the transcurrent motion between the Pacific and North America plates. We propose that oceanic microplate fragmentation is driven by slab stretching, thus reorganizing the mantle flow around the slab edge and further contributing to slab weakening and increase in buoyancy, eventually leading to cessation of subduction and microplate capture.

Device considerations for development of conductance-based biosensors

PubMed Central

Lee, Kangho; Nair, Pradeep R.; Scott, Adina; Alam, Muhammad A.; Janes, David B.

2009-01-01

Design and fabrication of electronic biosensors based on field-effect-transistor (FET) devices require understanding of interactions between semiconductor surfaces and organic biomolecules. From this perspective, we review practical considerations for electronic biosensors with emphasis on molecular passivation effects on FET device characteristics upon immobilization of organic molecules and an electrostatic model for FET-based biosensors. PMID:24753627

Glucocorticoid receptor ligand binding in monocytic cells using a microplate assay.

PubMed

Jansen, J; Uitdehaag, B; Koper, J W; van Den Berg, T K

1999-01-01

Glucocorticoids have profound effects on macrophage function and are widely used as anti-inflammatory drugs. Glucocorticoids receptor (GR) ligand binding capacity is a major determinant of cellular glucocorticoid sensitivity. The number and affinity of GR can be measured in a whole cell binding assay using (3)H-dexamethasone. Here, we describe a rapid and simple microplate assay for GR measurement using the human promonocytic cell line THP-1. Copyright 2000 S. Karger AG, Basel.

Fabric Organic Electrochemical Transistors for Biosensors.

PubMed

Yang, Anneng; Li, Yuanzhe; Yang, Chenxiao; Fu, Ying; Wang, Naixiang; Li, Li; Yan, Feng

2018-06-01

Flexible fabric biosensors can find promising applications in wearable electronics. However, high-performance fabric biosensors have been rarely reported due to many special requirements in device fabrication. Here, the preparation of organic electrochemical transistors (OECTs) on Nylon fibers is reported. By introducing metal/conductive polymer multilayer electrodes on the fibers, the OECTs show very stable performance during bending tests. The devices with functionalized gates are successfully used as various biosensors with high sensitivity and selectivity. The fiber-based OECTs are woven together with cotton yarns successfully by using a conventional weaving machine, resulting in flexible and stretchable fabric biosensors with high performance. The fabric sensors show much more stable signals in the analysis of moving aqueous solutions than planar devices due to a capillary effect in fabrics. The fabric devices are integrated in a diaper and remotely operated by using a mobile phone, offering a unique platform for convenient wearable healthcare monitoring. © 2018 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

A modified method for determining tannin-protein precipitation capacity using accelerated solvent extraction (ASE) and microplate gel filtration.

PubMed

McArt, Scott H; Spalinger, Donald E; Kennish, John M; Collins, William B

2006-06-01

The protein precipitation assay used by Robbins et al., (1987) Ecology 68:98-107 has been shown to predict successfully the reduction in protein availability to some ruminants due to tannins. The procedure, however, is expensive and laborious, which limits its utility, especially for quantitative ecological or nutritional applications where large numbers of assays may be required. We have modified the method to decrease its cost and increase laboratory efficiency by: (1) automating the extraction by using Accelerated Solvent Extraction (ASE); and (2) by scaling and automating the precipitation reaction, chromatography, and spectrometry with microplate gel filtration and an automated UV-VIS microplate spectrometer. ASE extraction is shown to be as effective at extracting tannins as the hot methanol technique. Additionally, the microplate assay is sensitive and precise. We show that the results from the new technique correspond in a nearly 1:1 relationship to the results of the previous technique. Hence, this method could reliably replace the older method with no loss in relevance to herbivore protein digestion. Moreover, the ASE extraction technique should be applicable to other tannin-protein precipitation assays and possibly other phenolic assays.

DNA Nanotechnology-Enabled Interfacial Engineering for Biosensor Development.

PubMed

Ye, Dekai; Zuo, Xiaolei; Fan, Chunhai

2018-06-12

Biosensors represent biomimetic analytical tools for addressing increasing needs in medical diagnosis, environmental monitoring, security, and biodefense. Nevertheless, widespread real-world applications of biosensors remain challenging due to limitations of performance, including sensitivity, specificity, speed, and reproducibility. In this review, we present a DNA nanotechnology-enabled interfacial engineering approach for improving the performance of biosensors. We first introduce the main challenges of the biosensing interfaces, especially under the context of controlling the DNA interfacial assembly. We then summarize recent progress in DNA nanotechnology and efforts to harness DNA nanostructures to engineer various biological interfaces, with a particular focus on the use of framework nucleic acids. We also discuss the implementation of biosensors to detect physiologically relevant nucleic acids, proteins, small molecules, ions, and other biomarkers. This review highlights promising applications of DNA nanotechnology in interfacial engineering for biosensors and related areas.

Superior Sensitivity of Copper-Based Plasmonic Biosensors.

PubMed

Stebunov, Yury V; Yakubovsky, Dmitry I; Fedyanin, Dmitry Yu; Arsenin, Aleksey V; Volkov, Valentyn S

2018-04-17

Plasmonic biosensing has been demonstrated to be a powerful technique for quantitative determination of molecular analytes and kinetic analysis of biochemical reactions. However, interfaces of most plasmonic biosensors are made of noble metals, such as gold and silver, which are not compatible with industrial production technologies. This greatly limits biosensing applications beyond biochemical and pharmaceutical research. Here, we propose and investigate copper-based biosensor chips fully fabricated with a standard complementary metal-oxide-semiconductor (CMOS) process. The protection of thin copper films from oxidation is achieved with SiO 2 and Al 2 O 3 dielectric films deposited onto the metal surface. In addition, the deposition of dielectric films with thicknesses of only several tens of nanometers significantly improves the biosensing sensitivity, owing to better localization of electromagnetic field above the biosensing surface. According to surface plasmon resonance (SPR) measurements, the copper biosensor chips coated with thin films of SiO 2 (25 nm) and Al 2 O 3 (15 nm) show 55% and 75% higher sensitivity to refractive index changes, respectively, in comparison to pure gold sensor chips. To test biomolecule immobilization, the copper-dielectric biosensor chips are coated with graphene oxide linking layers and used for the selective analysis of oligonucleotide hybridization. The proposed plasmonic biosensors make SPR technology more affordable for various applications and provide the basis for compact biosensors integrated with modern electronic devices.

Biosensors based on β-galactosidase enzyme: Recent advances and perspectives.

PubMed

Sharma, Shiv K; Leblanc, Roger M

2017-10-15

Many industries are striving for the development of more reliable and robust β-galactosidase biosensors that exhibit high response rate, increased detection limit and enriched useful lifetime. In a newfangled technological atmosphere, a trivial advantage or disadvantage of the developed biosensor may escort to the survival and extinction of the industry. Several alternative strategies to immobilize β-galactosidase enzyme for their utilization in biosensors have been developed in recent years in the quest of maximum utility by controlling the defects seen in the previous biosensors. The overwhelming call for on-line measurement of different sample constituents has directed science and industry to search for best practical solutions and biosensors are witnessed as the best prospect. The main objective of this paper is to serve as a narrow footbridge by comparing the literary works on the β-galactosidase biosensors, critically analyze their use in the construction of best biosensor by showing the pros and cons of the predicted methods for the practical use of biosensors. Copyright © 2017 Elsevier Inc. All rights reserved.

Modeling microelectrode biosensors: free-flow calibration can substantially underestimate tissue concentrations

PubMed Central

Wall, Mark J.

2016-01-01

Microelectrode amperometric biosensors are widely used to measure concentrations of analytes in solution and tissue including acetylcholine, adenosine, glucose, and glutamate. A great deal of experimental and modeling effort has been directed at quantifying the response of the biosensors themselves; however, the influence that the macroscopic tissue environment has on biosensor response has not been subjected to the same level of scrutiny. Here we identify an important issue in the way microelectrode biosensors are calibrated that is likely to have led to underestimations of analyte tissue concentrations. Concentration in tissue is typically determined by comparing the biosensor signal to that measured in free-flow calibration conditions. In a free-flow environment the concentration of the analyte at the outer surface of the biosensor can be considered constant. However, in tissue the analyte reaches the biosensor surface by diffusion through the extracellular space. Because the enzymes in the biosensor break down the analyte, a density gradient is set up resulting in a significantly lower concentration of analyte near the biosensor surface. This effect is compounded by the diminished volume fraction (porosity) and reduction in the diffusion coefficient due to obstructions (tortuosity) in tissue. We demonstrate this effect through modeling and experimentally verify our predictions in diffusive environments. NEW & NOTEWORTHY Microelectrode biosensors are typically calibrated in a free-flow environment where the concentrations at the biosensor surface are constant. However, when in tissue, the analyte reaches the biosensor via diffusion and so analyte breakdown by the biosensor results in a concentration gradient and consequently a lower concentration around the biosensor. This effect means that naive free-flow calibration will underestimate tissue concentration. We develop mathematical models to better quantify the discrepancy between the calibration and tissue

Modeling microelectrode biosensors: free-flow calibration can substantially underestimate tissue concentrations.

PubMed

Newton, Adam J H; Wall, Mark J; Richardson, Magnus J E

2017-03-01

Microelectrode amperometric biosensors are widely used to measure concentrations of analytes in solution and tissue including acetylcholine, adenosine, glucose, and glutamate. A great deal of experimental and modeling effort has been directed at quantifying the response of the biosensors themselves; however, the influence that the macroscopic tissue environment has on biosensor response has not been subjected to the same level of scrutiny. Here we identify an important issue in the way microelectrode biosensors are calibrated that is likely to have led to underestimations of analyte tissue concentrations. Concentration in tissue is typically determined by comparing the biosensor signal to that measured in free-flow calibration conditions. In a free-flow environment the concentration of the analyte at the outer surface of the biosensor can be considered constant. However, in tissue the analyte reaches the biosensor surface by diffusion through the extracellular space. Because the enzymes in the biosensor break down the analyte, a density gradient is set up resulting in a significantly lower concentration of analyte near the biosensor surface. This effect is compounded by the diminished volume fraction (porosity) and reduction in the diffusion coefficient due to obstructions (tortuosity) in tissue. We demonstrate this effect through modeling and experimentally verify our predictions in diffusive environments. NEW & NOTEWORTHY Microelectrode biosensors are typically calibrated in a free-flow environment where the concentrations at the biosensor surface are constant. However, when in tissue, the analyte reaches the biosensor via diffusion and so analyte breakdown by the biosensor results in a concentration gradient and consequently a lower concentration around the biosensor. This effect means that naive free-flow calibration will underestimate tissue concentration. We develop mathematical models to better quantify the discrepancy between the calibration and tissue

BIOSENSORS FOR ENVIRONMENTAL MONITORING: A REGULATORY PERSPECTIVE

EPA Science Inventory

Biosensors show the potential to complement laboratory-based analytical methods for environmental applications. Although biosensors for potential environmental-monitoring applications have been reported for a wide range of environmental pollutants, from a regulatory perspective, ...

Large Scale Bacterial Colony Screening of Diversified FRET Biosensors

PubMed Central

Litzlbauer, Julia; Schifferer, Martina; Ng, David; Fabritius, Arne; Thestrup, Thomas; Griesbeck, Oliver

2015-01-01

Biosensors based on Förster Resonance Energy Transfer (FRET) between fluorescent protein mutants have started to revolutionize physiology and biochemistry. However, many types of FRET biosensors show relatively small FRET changes, making measurements with these probes challenging when used under sub-optimal experimental conditions. Thus, a major effort in the field currently lies in designing new optimization strategies for these types of sensors. Here we describe procedures for optimizing FRET changes by large scale screening of mutant biosensor libraries in bacterial colonies. We describe optimization of biosensor expression, permeabilization of bacteria, software tools for analysis, and screening conditions. The procedures reported here may help in improving FRET changes in multiple suitable classes of biosensors. PMID:26061878

Biosensor method and system based on feature vector extraction

DOEpatents

Greenbaum, Elias [Knoxville, TN; Rodriguez, Jr., Miguel; Qi, Hairong [Knoxville, TN; Wang, Xiaoling [San Jose, CA

2012-04-17

A method of biosensor-based detection of toxins comprises the steps of providing at least one time-dependent control signal generated by a biosensor in a gas or liquid medium, and obtaining a time-dependent biosensor signal from the biosensor in the gas or liquid medium to be monitored or analyzed for the presence of one or more toxins selected from chemical, biological or radiological agents. The time-dependent biosensor signal is processed to obtain a plurality of feature vectors using at least one of amplitude statistics and a time-frequency analysis. At least one parameter relating to toxicity of the gas or liquid medium is then determined from the feature vectors based on reference to the control signal.

Biosensor method and system based on feature vector extraction

DOEpatents

Greenbaum, Elias; Rodriguez, Jr., Miguel; Qi, Hairong; Wang, Xiaoling

2013-07-02

A system for biosensor-based detection of toxins includes providing at least one time-dependent control signal generated by a biosensor in a gas or liquid medium, and obtaining a time-dependent biosensor signal from the biosensor in the gas or liquid medium to be monitored or analyzed for the presence of one or more toxins selected from chemical, biological or radiological agents. The time-dependent biosensor signal is processed to obtain a plurality of feature vectors using at least one of amplitude statistics and a time-frequency analysis. At least one parameter relating to toxicity of the gas or liquid medium is then determined from the feature vectors based on reference to the control signal.

Emerging Synergy between Nanotechnology and Implantable Biosensors: A Review

PubMed Central

Vaddiraju, Santhisagar; Tomazos, Ioannis; Burgess, Diane J; Jain, Faquir C; Papadimitrakopoulos, Fotios

2010-01-01

The development of implantable biosensors for continuous monitoring of metabolites is an area of sustained scientific and technological interest. On the other hand, nanotechnology, a discipline which deals with the properties of materials at the nanoscale, is developing as a potent tool to enhance the performance of these biosensors. This article reviews the current state of implantable biosensors, highlighting the synergy between nanotechnology and sensor performance. Emphasis is placed on the electrochemical method of detection in light of its widespread usage and substantial nanotechnology-based improvements in various aspects of electrochemical biosensor performance. Finally, issues regarding toxicity and biocompatibility of nanomaterials, along with future prospects for the application of nanotechnology in implantable biosensors, are discussed. PMID:20042326

Design Strategies for Aptamer-Based Biosensors

PubMed Central

Han, Kun; Liang, Zhiqiang; Zhou, Nandi

2010-01-01

Aptamers have been widely used as recognition elements for biosensor construction, especially in the detection of proteins or small molecule targets, and regarded as promising alternatives for antibodies in bioassay areas. In this review, we present an overview of reported design strategies for the fabrication of biosensors and classify them into four basic modes: target-induced structure switching mode, sandwich or sandwich-like mode, target-induced dissociation/displacement mode and competitive replacement mode. In view of the unprecedented advantages brought about by aptamers and smart design strategies, aptamer-based biosensors are expected to be one of the most promising devices in bioassay related applications. PMID:22399891

Bioelectrochemical interface engineering: toward the fabrication of electrochemical biosensors, biofuel cells, and self-powered logic biosensors.

PubMed

Zhou, Ming; Dong, Shaojun

2011-11-15

Over the past decade, researchers have devoted considerable attention to the integration of living organisms with electronic elements to yield bioelectronic devices. Not only is the integration of DNA, enzymes, or whole cells with electronics of scientific interest, but it has many versatile potential applications. Researchers are using these ideas to fabricate biosensors for analytical applications and to assemble biofuel cells (BFCs) and biomolecule-based devices. Other research efforts include the development of biocomputing systems for information processing. In this Account, we focus on our recent progress in engineering at the bioelectrochemical interface (BECI) for the rational design and construction of important bioelectronic devices, ranging from electrochemical (EC-) biosensors to BFCs, and self-powered logic biosensors. Hydrogels and sol-gels provide attractive materials for the immobilization of enzymes because they make EC-enzyme biosensors stable and even functional in extreme environments. We use a layer-by-layer (LBL) self-assembly technique to fabricate multicomponent thin films on the BECI at the nanometer scale. Additionally, we demonstrate how carbon nanomaterials have paved the way for new and improved EC-enzyme biosensors. In addition to the widely reported BECI-based electrochemical impedance spectroscopy (EIS)-type aptasensors, we integrate the LBL technique with our previously developed "solid-state probe" technique for redox probes immobilization on electrode surfaces to design and fabricate BECI-based differential pulse voltammetry (DPV)-type aptasensors. BFCs can directly harvest energy from ambient biofuels as green energy sources, which could lead to their application as simple, flexible, and portable power sources. Porous materials provide favorable microenvironments for enzyme immobilization, which can enhance BFC power output. Furthermore, by introducing aptamer-based logic systems to BFCs, such systems could be applied as self

Optical fiber-based biosensors.

PubMed

Monk, David J; Walt, David R

2004-08-01

This review outlines optical fiber-based biosensor research from January 2001 through September 2003 and was written to complement the previous review in this journal by Marazuela and Moreno-Bondi. Optical fiber-based biosensors combine the use of a biological recognition element with an optical fiber or optical fiber bundle. They are classified by the nature of the biological recognition element used for sensing: enzyme, antibody/antigen (immunoassay), nucleic acid, whole cell, and biomimetic, and may be used for a variety of analytes ranging from metals and chemicals to physiological materials.

Recent Developments in Enzyme, DNA and Immuno-Based Biosensors.

PubMed

Asal, Melis; Özen, Özlem; Şahinler, Mert; Polatoğlu, İlker

2018-06-13

Novel sensitive, rapid and economical biosensors are being developed in a wide range of medical environmental and food applications. In this paper, we review some of the main advances in the field over the past few years by discussing recent studies from literature. A biosensor, which is defined as an analytical device consisting of a biomolecule, a transducer and an output system, can be categorized according to the type of the incorporated biomolecule. The biomolecules can be enzymes, antibodies, ssDNA, organelles, cells etc. The main biosensor categories classified according to the biomolecules are enzymatic biosensors, immunosensors and DNA-based biosensors. These sensors can measure analytes produced or reduced during reactions at lower costs compared to the conventional detection techniques. Numerous types of biosensor studies conducted over the last decade have been explored here to reveal their key applications in medical, environmental and food industries which provide comprehensive perspective to the readers. Overviews of the working principles and applications of the reviewed sensors are also summarized.

Fundamental Design Principles for Transcription-Factor-Based Metabolite Biosensors.

PubMed

Mannan, Ahmad A; Liu, Di; Zhang, Fuzhong; Oyarzún, Diego A

2017-10-20

Metabolite biosensors are central to current efforts toward precision engineering of metabolism. Although most research has focused on building new biosensors, their tunability remains poorly understood and is fundamental for their broad applicability. Here we asked how genetic modifications shape the dose-response curve of biosensors based on metabolite-responsive transcription factors. Using the lac system in Escherichia coli as a model system, we built promoter libraries with variable operator sites that reveal interdependencies between biosensor dynamic range and response threshold. We developed a phenomenological theory to quantify such design constraints in biosensors with various architectures and tunable parameters. Our theory reveals a maximal achievable dynamic range and exposes tunable parameters for orthogonal control of dynamic range and response threshold. Our work sheds light on fundamental limits of synthetic biology designs and provides quantitative guidelines for biosensor design in applications such as dynamic pathway control, strain optimization, and real-time monitoring of metabolism.

Biosensors in the small scale: methods and technology trends.

PubMed

Senveli, Sukru U; Tigli, Onur

2013-03-01

This study presents a review on biosensors with an emphasis on recent developments in the field. A brief history accompanied by a detailed description of the biosensor concepts is followed by rising trends observed in contemporary micro- and nanoscale biosensors. Performance metrics to quantify and compare different detection mechanisms are presented. A comprehensive analysis on various types and subtypes of biosensors are given. The fields of interest within the scope of this review are label-free electrical, mechanical and optical biosensors as well as other emerging and popular technologies. Especially, the latter half of the last decade is reviewed for the types, methods and results of the most prominently researched detection mechanisms. Tables are provided for comparison of various competing technologies in the literature. The conclusion part summarises the noteworthy advantages and disadvantages of all biosensors reviewed in this study. Furthermore, future directions that the micro- and nanoscale biosensing technologies are expected to take are provided along with the immediate outlook.

Emerging synergy between nanotechnology and implantable biosensors: a review.

PubMed

Vaddiraju, Santhisagar; Tomazos, Ioannis; Burgess, Diane J; Jain, Faquir C; Papadimitrakopoulos, Fotios

2010-03-15

The development of implantable biosensors for continuous monitoring of metabolites is an area of sustained scientific and technological interests. On the other hand, nanotechnology, a discipline which deals with the properties of materials at the nanoscale, is developing as a potent tool to enhance the performance of these biosensors. This article reviews the current state of implantable biosensors, highlighting the synergy between nanotechnology and sensor performance. Emphasis is placed on the electrochemical method of detection in light of its widespread usage and substantial nanotechnology based improvements in various aspects of electrochemical biosensor performance. Finally, issues regarding toxicity and biocompatibility of nanomaterials, along with future prospects for the application of nanotechnology in implantable biosensors, are discussed. (c) 2009 Elsevier B.V. All rights reserved.

Fiber optic-based regenerable biosensor

DOEpatents

Sepaniak, Michael J.; Vo-Dinh, Tuan

1993-01-01

A fiber optic-based regenerable biosensor. The biosensor is particularly suitable for use in microscale work in situ. In one embodiment, the biosensor comprises a reaction chamber disposed adjacent the distal end of a waveguide and adapted to receive therein a quantity of a sample containing an analyte. Leading into the chamber is a plurality of capillary conduits suitable for introducing into the chamber antibodies or other reagents suitable for selective interaction with a predetermined analyte. Following such interaction, the contents of the chamber may be subjected to an incident energy signal for developing fluorescence within the chamber that is detectable via the optical fiber and which is representative of the presence, i.e. concentration, of the selected analyte. Regeneration of the biosensor is accomplished by replacement of the reagents and/or the analyte, or a combination of these, at least in part via one or more of the capillary conduits. The capillary conduits extend from their respective terminal ends that are in fluid communication with the chamber, away from the chamber to respective location(s) remote from the chamber thereby permitting in situ location of the chamber and remote manipulation and/or analysis of the activity with the chamber.

Biofuel metabolic engineering with biosensors.

PubMed

Morgan, Stacy-Anne; Nadler, Dana C; Yokoo, Rayka; Savage, David F

2016-12-01

Metabolic engineering offers the potential to renewably produce important classes of chemicals, particularly biofuels, at an industrial scale. DNA synthesis and editing techniques can generate large pathway libraries, yet identifying the best variants is slow and cumbersome. Traditionally, analytical methods like chromatography and mass spectrometry have been used to evaluate pathway variants, but such techniques cannot be performed with high throughput. Biosensors - genetically encoded components that actuate a cellular output in response to a change in metabolite concentration - are therefore a promising tool for rapid and high-throughput evaluation of candidate pathway variants. Applying biosensors can also dynamically tune pathways in response to metabolic changes, improving balance and productivity. Here, we describe the major classes of biosensors and briefly highlight recent progress in applying them to biofuel-related metabolic pathway engineering. Copyright © 2016 Elsevier Ltd. All rights reserved.

Development of a biosensor for caffeine.

PubMed

Babu, V R Sarath; Patra, S; Karanth, N G; Kumar, M A; Thakur, M S

2007-01-23

We have utilized a microbe, which can degrade caffeine to develop an Amperometric biosensor for determination of caffeine in solutions. Whole cells of Pseudomonas alcaligenes MTCC 5264 having the capability to degrade caffeine were immobilized on a cellophane membrane with a molecular weight cut off (MWCO) of 3000-6000 by covalent crosslinking method using glutaraledhyde as the bifunctional crosslinking agent and gelatin as the protein based stabilizing agent (PBSA). The biosensor system was able to detect caffeine in solution over a concentration range of 0.1 to 1 mg mL(-1). With read-times as short as 3 min, this caffeine biosensor acts as a rapid analysis system for caffeine in solutions. Interestingly, successful isolation and immobilization of caffeine degrading bacteria for the analysis of caffeine described here was enabled by a novel selection strategy that incorporated isolation of caffeine degrading bacteria capable of utilizing caffeine as the sole source of carbon and nitrogen from soils and induction of caffeine degrading capacity in bacteria for the development of the biosensor. This biosensor is highly specific for caffeine and response to interfering compounds such as theophylline, theobromine, paraxanthine, other methyl xanthines and sugars was found to be negligible. Although a few biosensing methods for caffeine are reported, they have limitations in application for commercial samples. The development and application of new caffeine detection methods remains an active area of investigation, particularly in food and clinical chemistry. The optimum pH and temperature of measurement were 6.8 and 30+/-2 degrees C, respectively. Interference in analysis of caffeine due to different substrates was observed but was not considerable. Caffeine content of commercial samples of instant tea and coffee was analyzed by the biosensor and the results compared well with HPLC analysis.

A novel biosensor selective for organoarsenicals.

PubMed

Chen, Jian; Zhu, Yong-Guan; Rosen, Barry P

2012-10-01

Organoarsenicals used as herbicides and growth promoters for farm animals are degraded to inorganic arsenic. Available bacterial whole-cell biosensors detect only inorganic arsenic. We report a biosensor selective for the trivalent organoarsenicals methylarsenite and phenylarsenite over inorganic arsenite. This sensor may be useful for detecting degradation of arsenic-containing herbicides and growth promoters.

Recent development of nano-materials used in DNA biosensors.

PubMed

Xu, Kai; Huang, Junran; Ye, Zunzhong; Ying, Yibin; Li, Yanbin

2009-01-01

As knowledge of the structure and function of nucleic acid molecules has increased, sequence-specific DNA detection has gained increased importance. DNA biosensors based on nucleic acid hybridization have been actively developed because of their specificity, speed, portability, and low cost. Recently, there has been considerable interest in using nano-materials for DNA biosensors. Because of their high surface-to-volume ratios and excellent biological compatibilities, nano-materials could be used to increase the amount of DNA immobilization; moreover, DNA bound to nano-materials can maintain its biological activity. Alternatively, signal amplification by labeling a targeted analyte with nano-materials has also been reported for DNA biosensors in many papers. This review summarizes the applications of various nano-materials for DNA biosensors during past five years. We found that nano-materials of small sizes were advantageous as substrates for DNA attachment or as labels for signal amplification; and use of two or more types of nano-materials in the biosensors could improve their overall quality and to overcome the deficiencies of the individual nano-components. Most current DNA biosensors require the use of polymerase chain reaction (PCR) in their protocols. However, further development of nano-materials with smaller size and/or with improved biological and chemical properties would substantially enhance the accuracy, selectivity and sensitivity of DNA biosensors. Thus, DNA biosensors without PCR amplification may become a reality in the foreseeable future.

Recent Development of Nano-Materials Used in DNA Biosensors

PubMed Central

Xu, Kai; Huang, Junran; Ye, Zunzhong; Ying, Yibin; Li, Yanbin

2009-01-01

As knowledge of the structure and function of nucleic acid molecules has increased, sequence-specific DNA detection has gained increased importance. DNA biosensors based on nucleic acid hybridization have been actively developed because of their specificity, speed, portability, and low cost. Recently, there has been considerable interest in using nano-materials for DNA biosensors. Because of their high surface-to-volume ratios and excellent biological compatibilities, nano-materials could be used to increase the amount of DNA immobilization; moreover, DNA bound to nano-materials can maintain its biological activity. Alternatively, signal amplification by labeling a targeted analyte with nano-materials has also been reported for DNA biosensors in many papers. This review summarizes the applications of various nano-materials for DNA biosensors during past five years. We found that nano-materials of small sizes were advantageous as substrates for DNA attachment or as labels for signal amplification; and use of two or more types of nano-materials in the biosensors could improve their overall quality and to overcome the deficiencies of the individual nano-components. Most current DNA biosensors require the use of polymerase chain reaction (PCR) in their protocols. However, further development of nano-materials with smaller size and/or with improved biological and chemical properties would substantially enhance the accuracy, selectivity and sensitivity of DNA biosensors. Thus, DNA biosensors without PCR amplification may become a reality in the foreseeable future. PMID:22346713

Nanoscale bacteriophage biosensors beyond phage display.

PubMed

Lee, Jong-Wook; Song, Jangwon; Hwang, Mintai P; Lee, Kwan Hyi

2013-01-01

Bacteriophages are traditionally used for the development of phage display technology. Recently, their nanosized dimensions and ease with which genetic modifications can be made to their structure and function have put them in the spotlight towards their use in a variety of biosensors. In particular, the expression of any protein or peptide on the extraluminal surface of bacteriophages is possible by genetically engineering the genome. In addition, the relatively short replication time of bacteriophages offers researchers the ability to generate mass quantities of any given bacteriophage-based biosensor. Coupled with the emergence of various biomarkers in the clinic as a means to determine pathophysiological states, the development of current and novel technologies for their detection and quantification is imperative. In this review, we categorize bacteriophages by their morphology into M13-based filamentous bacteriophages and T4- or T7-based icosahedral bacteriophages, and examine how such advantages are utilized across a variety of biosensors. In essence, we take a comprehensive approach towards recent trends in bacteriophage-based biosensor applications and discuss their outlook with regards to the field of biotechnology.

Nanoscale bacteriophage biosensors beyond phage display

PubMed Central

Lee, Jong-Wook; Song, Jangwon; Hwang, Mintai P; Lee, Kwan Hyi

2013-01-01

Bacteriophages are traditionally used for the development of phage display technology. Recently, their nanosized dimensions and ease with which genetic modifications can be made to their structure and function have put them in the spotlight towards their use in a variety of biosensors. In particular, the expression of any protein or peptide on the extraluminal surface of bacteriophages is possible by genetically engineering the genome. In addition, the relatively short replication time of bacteriophages offers researchers the ability to generate mass quantities of any given bacteriophage-based biosensor. Coupled with the emergence of various biomarkers in the clinic as a means to determine pathophysiological states, the development of current and novel technologies for their detection and quantification is imperative. In this review, we categorize bacteriophages by their morphology into M13-based filamentous bacteriophages and T4- or T7-based icosahedral bacteriophages, and examine how such advantages are utilized across a variety of biosensors. In essence, we take a comprehensive approach towards recent trends in bacteriophage-based biosensor applications and discuss their outlook with regards to the field of biotechnology. PMID:24143096

Modularization and Response Curve Engineering of a Naringenin-Responsive Transcriptional Biosensor.

PubMed

De Paepe, Brecht; Maertens, Jo; Vanholme, Bartel; De Mey, Marjan

2018-05-18

To monitor the intra- and extracellular environment of micro-organisms and to adapt their metabolic processes accordingly, scientists are reprogramming nature's myriad of transcriptional regulatory systems into transcriptional biosensors, which are able to detect small molecules and, in response, express specific output signals of choice. However, the naturally occurring response curve, the key characteristic of biosensor circuits, is typically not in line with the requirements for real-life biosensor applications. In this contribution, a natural LysR-type naringenin-responsive biosensor circuit is developed and characterized with Escherichia coli as host organism. Subsequently, this biosensor is dissected into a clearly defined detector and effector module without loss of functionality, and the influence of the expression levels of both modules on the biosensor response characteristics is investigated. Two collections of ten unique synthetic biosensors each are generated. Each collection demonstrates a unique diversity of response curve characteristics spanning a 128-fold change in dynamic and 2.5-fold change in operational ranges and 3-fold change in levels of Noise, fit for a wide range of applications, such as adaptive laboratory evolution, dynamic pathway control and high-throughput screening methods. The established biosensor engineering concepts, and the developed biosensor collections themselves, are of use for the future development and customization of biosensors in general, for the multitude of biosensor applications and as a compelling alternative for the commonly used LacI-, TetR- and AraC-based inducible circuits.

Rapid development of new protein biosensors utilizing peptides obtained via phage display.

PubMed

Wu, Jun; Park, Jong Pil; Dooley, Kevin; Cropek, Donald M; West, Alan C; Banta, Scott

2011-01-01

There is a consistent demand for new biosensors for the detection of protein targets, and a systematic method for the rapid development of new sensors is needed. Here we present a platform where short unstructured peptides that bind to a desired target are selected using M13 phage display. The selected peptides are then chemically synthesized and immobilized on gold, allowing for detection of the target using electrochemical techniques such as electrochemical impedance spectroscopy (EIS). A quartz crystal microbalance (QCM) is also used as a diagnostic tool during biosensor development. We demonstrate the utility of this approach by creating a novel peptide-based electrochemical biosensor for the enzyme alanine aminotransferase (ALT), a well-known biomarker of hepatotoxicity. Biopanning of the M13 phage display library over immobilized ALT, led to the rapid identification of a new peptide (ALT5-8) with an amino acid sequence of WHWRNPDFWYLK. Phage particles expressing this peptide exhibited nanomolar affinity for immobilized ALT (K(d,app) = 85±20 nM). The newly identified ALT5-8 peptide was then chemically synthesized with a C-terminal cysteine for gold immobilization. The performance of the gold-immobilized peptides was studied with cyclic voltammetry (CV), QCM, and EIS. Using QCM, the sensitivity for ALT detection was 8.9±0.9 Hz/(µg/mL) and the limit of detection (LOD) was 60 ng/mL. Using EIS measurements, the sensitivity was 142±12 impedance percentage change %/(µg/mL) and the LOD was 92 ng/mL. In both cases, the LOD was below the typical concentration of ALT in human blood. Although both QCM and EIS produced similar LODs, EIS is preferable due to a larger linear dynamic range. Using QCM, the immobilized peptide exhibited a nanomolar dissociation constant for ALT (K(d) = 20.1±0.6 nM). These results demonstrate a simple and rapid platform for developing and assessing the performance of sensitive, peptide-based biosensors for new protein targets.

Highly sensitive quartz crystal microbalance based biosensor using Au dendrite structure

NASA Astrophysics Data System (ADS)

Asai, Naoto; Terasawa, Hideaki; Shimizu, Tomohiro; Shingubara, Shoso; Ito, Takeshi

2018-02-01

A Au dendrite structure was obtained by only electroplating under a suitable potential. A blanch like nanostructure was formed along the crystal orientation. In this study, we attempted to fabricate a Au dendrite structure on the electrode of a quartz crystal by electroplating to increase the specific surface area. We estimated the effective surface area by cyclic voltammetry (CV) and monitored the frequency shift induced by antigen-antibody interaction by the quartz crystal microbalance (QCM) method. The dendrite structure with the largest surface area was formed under -0.95 V for 5 min. In the measurement of the antigen-antibody interaction, the frequency shifts of 40, 80, and 110 Hz were obtained with the dendrite structured QCM chips formed at the above potential for 1, 1.5, and 2.0 min, respectively. The sensitivity was improved compared with that QCM chip having a flat surface electrode.

Recent advances in polyaniline based biosensors.

PubMed

Dhand, Chetna; Das, Maumita; Datta, Monika; Malhotra, B D

2011-02-15

The present paper contains a detailed overview of recent advances relating to polyaniline (PANI) as a transducer material for biosensor applications. This conducting polymer provides enormous opportunities for binding biomolecules, tuning their bio-catalytic properties, rapid electron transfer and direct communication to produce a range of analytical signals and new analytical applications. Merging the specific nature of different biomolecules (enzymes, nucleic acids, antibodies, etc.) and the key properties of this modern conducting matrix, possible biosensor designs and their biosensing characteristics have been discussed. Efforts have been made to discuss and explore various characteristics of PANI responsible for direct electron transfer leading towards fabrication of mediator-less biosensors. Copyright © 2010 Elsevier B.V. All rights reserved.

Graphene-Based Optical Biosensors and Imaging

DOE Office of Scientific and Technical Information (OSTI.GOV)

Tang, Zhiwen; He, Shijiang; Pei, Hao

2014-01-13

This chapter focuses on the design, fabrication and application of graphene based optical nanobiosensors. The emerging graphene based optical nanobiosensors demonstrated the promising bioassay and biomedical applications thanking to the unique optical features of graphene. According to the different applications, the graphene can be tailored to form either fluorescent emitter or efficient fluorescence quencher. The exceptional electronic feature of graphene makes it a powerful platform for fabricating the SPR and SERS biosensors. Today the graphene based optical biosensors have been constructed to detect various targets including ions, small biomolecules, DNA/RNA and proteins. This chapter reviews the recent progress in graphene-basedmore » optical biosensors and discusses the opportunities and challenges in this field.« less

Microplate-based active/inactive 1 screen for biomass degrading enzyme library purification and gene discovery

USDA-ARS?s Scientific Manuscript database

We present here a whole-cell and permeabilized E. coli cell 1' active/inactive microplate screen for ß-D-xylosidase, xylanase, endocellulase, and ferulic acid esterase enzyme activities which are critical for the enzymatic deconstruction of biomass for fuels and chemicals. Transformants from genomic...

Revision of the genera Microplitis and Snellenius (Hymenoptera, Braconidae, Microgastrinae) from Area de Conservacion Guanacaste, Costa Rica, with a key to all species previously described from Mesoamerica

USDA-ARS?s Scientific Manuscript database

The genera Microplitis and Snellenius (Hymenoptera: Braconidae, Microgastrinae) from Area de Conservacion Guanacaste (ACG), Costa Rica, are revised. A total of 28 new species are described: 23 of Snellenius (the first record for Mesoamerica) and five of Microplitis. A key is provided to all new spec...

Solvent minimization induces preferential orientation and crystal clustering in serial micro-crystallography on micro-meshes, in situ plates and on a movable crystal conveyor belt

PubMed Central

Soares, Alexei S.; Mullen, Jeffrey D.; Parekh, Ruchi M.; McCarthy, Grace S.; Roessler, Christian G.; Jackimowicz, Rick; Skinner, John M.; Orville, Allen M.; Allaire, Marc; Sweet, Robert M.

2014-01-01

X-ray diffraction data were obtained at the National Synchrotron Light Source from insulin and lysozyme crystals that were densely deposited on three types of surfaces suitable for serial micro-crystallography: MiTeGen MicroMeshes™, Greiner Bio-One Ltd in situ micro-plates, and a moving kapton crystal conveyor belt that is used to deliver crystals directly into the X-ray beam. 6° wedges of data were taken from ∼100 crystals mounted on each material, and these individual data sets were merged to form nine complete data sets (six from insulin crystals and three from lysozyme crystals). Insulin crystals have a parallelepiped habit with an extended flat face that preferentially aligned with the mounting surfaces, impacting the data collection strategy and the design of the serial crystallography apparatus. Lysozyme crystals had a cuboidal habit and showed no preferential orientation. Preferential orientation occluded regions of reciprocal space when the X-ray beam was incident normal to the data-collection medium surface, requiring a second pass of data collection with the apparatus inclined away from the orthogonal. In addition, crystals measuring less than 20 µm were observed to clump together into clusters of crystals. Clustering required that the X-ray beam be adjusted to match the crystal size to prevent overlapping diffraction patterns. No additional problems were encountered with the serial crystallography strategy of combining small randomly oriented wedges of data from a large number of specimens. High-quality data able to support a realistic molecular replacement solution were readily obtained from both crystal types using all three serial crystallography strategies. PMID:25343789

Carbon nanomaterials in biosensors: should you use nanotubes or graphene?

PubMed

Yang, Wenrong; Ratinac, Kyle R; Ringer, Simon P; Thordarson, Pall; Gooding, J Justin; Braet, Filip

2010-03-15

From diagnosis of life-threatening diseases to detection of biological agents in warfare or terrorist attacks, biosensors are becoming a critical part of modern life. Many recent biosensors have incorporated carbon nanotubes as sensing elements, while a growing body of work has begun to do the same with the emergent nanomaterial graphene, which is effectively an unrolled nanotube. With this widespread use of carbon nanomaterials in biosensors, it is timely to assess how this trend is contributing to the science and applications of biosensors. This Review explores these issues by presenting the latest advances in electrochemical, electrical, and optical biosensors that use carbon nanotubes and graphene, and critically compares the performance of the two carbon allotropes in this application. Ultimately, carbon nanomaterials, although still to meet key challenges in fabrication and handling, have a bright future as biosensors.

A thermal biosensor based on enzyme reaction.

PubMed

Zheng, Yi-Hua; Hua, Tse-Chao; Xu, Fei

2005-01-01

Application of the thermal biosensor as analytical tool is promising due to advantages as universal, simplicity and quick response. A novel thermal biosensor based on enzyme reaction has been developed. This biosensor is a flow injection analysis system and consists of two channels with enzyme reaction column and reference column. The reference column, which is set for eliminating the unspecific heat, is inactived on special enzyme reaction of the ingredient to be detected. The special enzyme reaction takes places in the enzyme reaction column at a constant temperature realizing by a thermoelectric thermostat. Thermal sensor based on the thermoelectric module containing 127 serial BiTe-thermocouples is used to monitor the temperature difference between two streams from the enzyme reaction column and the reference column. The analytical example for dichlorvos shows that this biosensor can be used as analytical tool in medicine and biology.

Effect of Diffusion Limitations on Multianalyte Determination from Biased Biosensor Response

PubMed Central

Baronas, Romas; Kulys, Juozas; Lančinskas, Algirdas; Žilinskas, Antanas

2014-01-01

The optimization-based quantitative determination of multianalyte concentrations from biased biosensor responses is investigated under internal and external diffusion-limited conditions. A computational model of a biocatalytic amperometric biosensor utilizing a mono-enzyme-catalyzed (nonspecific) competitive conversion of two substrates was used to generate pseudo-experimental responses to mixtures of compounds. The influence of possible perturbations of the biosensor signal, due to a white noise- and temperature-induced trend, on the precision of the concentration determination has been investigated for different configurations of the biosensor operation. The optimization method was found to be suitable and accurate enough for the quantitative determination of the concentrations of the compounds from a given biosensor transient response. The computational experiments showed a complex dependence of the precision of the concentration estimation on the relative thickness of the outer diffusion layer, as well as on whether the biosensor operates under diffusion- or kinetics-limited conditions. When the biosensor response is affected by the induced exponential trend, the duration of the biosensor action can be optimized for increasing the accuracy of the quantitative analysis. PMID:24608006

Antibodies and antibody-derived analytical biosensors

PubMed Central

Sharma, Shikha; Byrne, Hannah

2016-01-01

The rapid diagnosis of many diseases and timely initiation of appropriate treatment are critical determinants that promote optimal clinical outcomes and general public health. Biosensors are now being applied for rapid diagnostics due to their capacity for point-of-care use with minimum need for operator input. Antibody-based biosensors or immunosensors have revolutionized diagnostics for the detection of a plethora of analytes such as disease markers, food and environmental contaminants, biological warfare agents and illicit drugs. Antibodies are ideal biorecognition elements that provide sensors with high specificity and sensitivity. This review describes monoclonal and recombinant antibodies and different immobilization approaches crucial for antibody utilization in biosensors. Examples of applications of a variety of antibody-based sensor formats are also described. PMID:27365031

A microplate assay to measure classical and alternative complement activity.

PubMed

Puissant-Lubrano, Bénédicte; Fortenfant, Françoise; Winterton, Peter; Blancher, Antoine

2017-05-01

We developed and validated a kinetic microplate hemolytic assay (HA) to quantify classical and alternative complement activity in a single dilution of human plasma or serum. The assay is based on monitoring hemolysis of sensitized sheep (or uncoated rabbit) red blood cells by means of a 96-well microplate reader. The activity of the calibrator was evaluated by reference to 200 healthy adults. The conversion of 50% hemolysis time into a percentage of activity was obtained using a calibration curve plotted daily. The linearity of the assay as well as interference (by hemolysis, bilrubinemia and lipemia) was assessed for classical pathway (CP). The within-day and the between-day precision was satisfactory regarding the performance of commercially available liposome immunoassay (LIA) and ELISA. Patients with hereditary or acquired complement deficiencies were detected (activity was measured <30%). We also provided a reference range obtained from 200 blood donors. The agreement of CP evaluated on samples from 48 patients was 94% with LIA and 87.5% with ELISA. The sensitivity of our assay was better than that of LIA, and the cost was lower than either LIA or ELISA. In addition, this assay was less time consuming than previously reported HAs. This assay allows the simultaneous measurement of 36 samples in duplicate per run of a 96-well plate. The use of a daily calibration curve allows standardization of the method and leads to good reproducibility. The same technique was also adapted for the quantification of alternative pathway (AP) activity.

Analytical parameters of the microplate-based ORAC-pyrogallol red assay.

PubMed

Ortiz, Rocío; Antilén, Mónica; Speisky, Hernán; Aliaga, Margarita E; López-Alarcón, Camilo

2011-01-01

The analytical parameters of the microplate-based oxygen radicals absorbance capacity (ORAC) method using pyrogallol red (PGR) as probe (ORAC-PGR) are presented. In addition, the antioxidant capacity of commercial beverages, such as wines, fruit juices, and iced teas, is estimated. A good linearity of the area under the curve (AUC) versus Trolox concentration plots was obtained [AUC = (845 +/- 110) + (23 +/- 2) [Trolox, microM], R = 0.9961, n = 19]. QC experiments showed better precision and accuracy at the highest Trolox concentration (40 microM) with RSD and REC (recuperation) values of 1.7 and 101.0%, respectively. When red wine was used as sample, the method also showed good linearity [AUC = (787 +/- 77) + (690 +/- 60) [red wine, microL/mL]; R = 0.9926, n = 17], precision and accuracy with RSD values from 1.4 to 8.3%, and REC values that ranged from 89.7 to 103.8%. Additivity assays using solutions containing gallic acid and Trolox (or red wine) showed an additive protection of PGR given by the samples. Red wines showed higher ORAC-PGR values than white wines, while the ORAC-PGR index of fruit juices and iced teas presented a great variability, ranging from 0.6 to 21.6 mM of Trolox equivalents. This variability was also observed for juices of the same fruit, showing the influence of the brand on the ORAC-PGR index. The ORAC-PGR methodology can be applied in a microplate reader with good linearity, precision, and accuracy.

Discrimination of peptides by using a molecularly imprinted piezoelectric biosensor.

PubMed

Lin, Chung-Yin; Tai, Dar-Fu; Wu, Tzong-Zeng

2003-10-17

Based on the direct formation of a molecularly imprinted polymer on gold electrodes, we have developed a peptide sensor for the detection of low-molecular-weight peptides. A new cross-linking monomer, (N-Acr-L-Cys-NHBn)(2), was employed to attach the surface of the chip and to copolymerize with other monomers. Interestingly, N-benzylacrylamide participates in the polymerization and recognition is carried out in an aqueous environment. By using quartz crystal microbalance detection, short peptides can be monitored by their interaction with plastic antibodies specific for the target peptides. The selectivity of molecularly imprinted polymers and the sensitivity of such artificial biosensors have been combined to differentiate between traces of oxytocin and vasopressin to the ng mL(-1) scale.

A general strategy to construct small molecule biosensors in eukaryotes.

PubMed

Feng, Justin; Jester, Benjamin W; Tinberg, Christine E; Mandell, Daniel J; Antunes, Mauricio S; Chari, Raj; Morey, Kevin J; Rios, Xavier; Medford, June I; Church, George M; Fields, Stanley; Baker, David

2015-12-29

Biosensors for small molecules can be used in applications that range from metabolic engineering to orthogonal control of transcription. Here, we produce biosensors based on a ligand-binding domain (LBD) by using a method that, in principle, can be applied to any target molecule. The LBD is fused to either a fluorescent protein or a transcriptional activator and is destabilized by mutation such that the fusion accumulates only in cells containing the target ligand. We illustrate the power of this method by developing biosensors for digoxin and progesterone. Addition of ligand to yeast, mammalian, or plant cells expressing a biosensor activates transcription with a dynamic range of up to ~100-fold. We use the biosensors to improve the biotransformation of pregnenolone to progesterone in yeast and to regulate CRISPR activity in mammalian cells. This work provides a general methodology to develop biosensors for a broad range of molecules in eukaryotes.

A living cell quartz crystal microbalance biosensor for continuous monitoring of cytotoxic responses of macrophages to single-walled carbon nanotubes

PubMed Central

2011-01-01

Background Numerous engineered nanomaterials (ENMs) exist and new ENMs are being developed. A challenge to nanotoxicology and environmental health and safety is evaluating toxicity of ENMs before they become widely utilized. Cellular assays remain the predominant test platform yet these methods are limited by using discrete time endpoints and reliance on organic dyes, vulnerable to interference from ENMs. Label-free, continuous, rapid response systems with biologically meaningful endpoints are needed. We have developed a device to detect and monitor in real time responses of living cells to ENMs. The device, a living cell quartz crystal microbalance biosensor (QCMB), uses macrophages adherent to a quartz crystal. The communal response of macrophages to treatments is monitored continuously as changes in crystal oscillation frequency (Δf). We report the ability of this QCMB to distinguish benign from toxic exposures and reveal unique kinetic information about cellular responses to varying doses of single-walled carbon nanotubes (SWCNTs). Results We analyzed macrophage responses to additions of Zymosan A, polystyrene beads (PBs) (benign substances) or SWCNT (3-150 μg/ml) in the QCMB over 18 hrs. In parallel, toxicity was monitored over 24/48 hrs using conventional viability assays and histological stains to detect apoptosis. In the QCMB, a stable unchanging oscillation frequency occurred when cells alone, Zymosan A alone, PBs alone or SWCNTs without cells at the highest dose alone were used. With living cells in the QCMB, when Zymosan A, PBs or SWCNTs were added, a significant decrease in frequency occurred from 1-6 hrs. For SWCNTs, this Δf was dose-dependent. From 6-18 hrs, benign substances or low dose SWCNT (3-30 μg/ml) treatments showed a reversal of the decrease of oscillation frequency, returning to or exceeding pre-treatment levels. Cell recovery was confirmed in conventional assays. The lag time to see the Δf reversal in QCMB plots was linearly SWCNT

In vitro chloramphenicol detection in a Haemophilus influenza model using an aptamer-polymer based electrochemical biosensor.

PubMed

Yadav, Saurabh K; Agrawal, Bharati; Chandra, Pranjal; Goyal, Rajendra N

2014-05-15

A sensitive and selective electrochemical biosensor is developed for the determination of chloramphenicol (CAP) exploring its direct electron transfer processes in in-vitro model and pharmaceutical samples. This biosensor exploits a selective binding of CAP with aptamer, immobilized onto the poly-(4-amino-3-hydroxynapthalene sulfonic acid) (p-AHNSA) modified edge plane pyrolytic graphite. The electrochemical reduction of CAP was observed in a well-defined peak. A quartz crystal microbalance (QCM) study is performed to confirm the interaction between the polymer film and the aptamer. Cyclic voltammetry (CV) and square wave voltammetry (SWV) were used to detect CAP. The in-vitro CAP detection is performed using the bacterial strain of Haemophilus influenza. A significant accumulation of CAP by the drug sensitive H. influenza strain is observed for the first time in this study using a biosensor. Various parameters affecting the CAP detection in standard solution and in in vitro detection are optimized. The detection of CAP is linear in the range of 0.1-2500 nM with the detection limit and sensitivity of 0.02 nM and 0.102 µA/nM, respectively. CAP is also detected in the presence of other common antibiotics and proteins present in the real sample matrix, and negligible interference is observed. Copyright © 2013 Elsevier B.V. All rights reserved.

Enzyme-linked, aptamer-based, competitive biolayer interferometry biosensor for palytoxin.

PubMed

Gao, Shunxiang; Zheng, Xin; Hu, Bo; Sun, Mingjuan; Wu, Jihong; Jiao, Binghua; Wang, Lianghua

2017-03-15

In this study, we coupled biolayer interferometry (BLI) with competitive binding assay through an enzyme-linked aptamer and developed a real-time, ultra-sensitive, rapid quantitative method for detection of the marine biotoxin palytoxin. Horseradish peroxidase-labeled aptamers were used as biorecognition receptors to competitively bind with palytoxin, which was immobilized on the biosensor surface. The palytoxin: horseradish peroxidase-aptamer complex was then submerged in a 3,3'-diaminobenzidine solution, which resulted in formation of a precipitated polymeric product directly on the biosensor surface and a large change in the optical thickness of the biosensor layer. This change could obviously shift the interference pattern and generate a response profile on the BLI biosensor. The biosensor showed a broad linear range for palytoxin (200-700pg/mL) with a low detection limit (0.04pg/mL). Moreover, the biosensor was applied to the detection of palytoxin in spiked extracts and showed a high degree of selectivity for palytoxin, good reproducibility, and stability. This enzyme-linked, aptamer-based, competitive BLI biosensor offers a promising method for rapid and sensitive detection of palytoxin and other analytes. Copyright © 2016 Elsevier B.V. All rights reserved.

Alginate cryogel based glucose biosensor

NASA Astrophysics Data System (ADS)

Fatoni, Amin; Windy Dwiasi, Dian; Hermawan, Dadan

2016-02-01

Cryogel is macroporous structure provides a large surface area for biomolecule immobilization. In this work, an alginate cryogel based biosensor was developed to detect glucose. The cryogel was prepared using alginate cross-linked by calcium chloride under sub-zero temperature. This porous structure was growth in a 100 μL micropipette tip with a glucose oxidase enzyme entrapped inside the cryogel. The glucose detection was based on the colour change of redox indicator, potassium permanganate, by the hydrogen peroxide resulted from the conversion of glucose. The result showed a porous structure of alginate cryogel with pores diameter of 20-50 μm. The developed glucose biosensor was showed a linear response in the glucose detection from 1.0 to 5.0 mM with a regression of y = 0.01x+0.02 and R2 of 0.994. Furthermore, the glucose biosensor was showed a high operational stability up to 10 times of uninterrupted glucose detections.

Magnetic biosensors: Modelling and simulation.

PubMed

Nabaei, Vahid; Chandrawati, Rona; Heidari, Hadi

2018-04-30

In the past few years, magnetoelectronics has emerged as a promising new platform technology in various biosensors for detection, identification, localisation and manipulation of a wide spectrum of biological, physical and chemical agents. The methods are based on the exposure of the magnetic field of a magnetically labelled biomolecule interacting with a complementary biomolecule bound to a magnetic field sensor. This Review presents various schemes of magnetic biosensor techniques from both simulation and modelling as well as analytical and numerical analysis points of view, and the performance variations under magnetic fields at steady and nonstationary states. This is followed by magnetic sensors modelling and simulations using advanced Multiphysics modelling software (e.g. Finite Element Method (FEM) etc.) and home-made developed tools. Furthermore, outlook and future directions of modelling and simulations of magnetic biosensors in different technologies and materials are critically discussed. Crown Copyright © 2017. Published by Elsevier B.V. All rights reserved.

Carbon nanotube biosensors

PubMed Central

Tîlmaciu, Carmen-Mihaela; Morris, May C.

2015-01-01

Nanomaterials possess unique features which make them particularly attractive for biosensing applications. In particular, carbon nanotubes (CNTs) can serve as scaffolds for immobilization of biomolecules at their surface, and combine several exceptional physical, chemical, electrical, and optical characteristics properties which make them one of the best suited materials for the transduction of signals associated with the recognition of analytes, metabolites, or disease biomarkers. Here we provide a comprehensive review on these carbon nanostructures, in which we describe their structural and physical properties, functionalization and cellular uptake, biocompatibility, and toxicity issues. We further review historical developments in the field of biosensors, and describe the different types of biosensors which have been developed over time, with specific focus on CNT-conjugates engineered for biosensing applications, and in particular detection of cancer biomarkers. PMID:26579509

Carbon Nanotube Biosensors

NASA Astrophysics Data System (ADS)

Tilmaciu, Carmen-Mihaela; Morris, May

2015-10-01

Nanomaterials possess unique features which make them particularly attractive for biosensing applications. In particular Carbon Nanotubes (CNTs) can serve as scaffolds for immobilization of biomolecules at their surface, and combine several exceptional physical, chemical, electrical and optical characteristics properties which make them one of the best suited materials for the transduction of signals associated with the recognition of analytes, metabolites or disease biomarkers. Here we provide a comprehensive review on these carbon nanostructures, in which we will describe their structural and physical properties, discuss functionalization and cellular uptake, biocompatibility and toxicity issues. We further review historical developments in the field of biosensors, and describe the different types of biosensors which have been developed over time, with specific focus on CNT-conjugates engineered for biosensing applications, and in particular detection of cancer biomarkers.

Applications of commercial biosensors in clinical, food, environmental, and biothreat/biowarfare analyses.

PubMed

Bahadır, Elif Burcu; Sezgintürk, Mustafa Kemal

2015-06-01

The lack of specific, low-cost, rapid, sensitive, and easy detection of biomolecules has resulted in the development of biosensor technology. Innovations in biosensor technology have enabled many biosensors to be commercialized and have enabled biomolecules to be detected onsite. Moreover, the emerging technologies of lab-on-a-chip microdevices and nanosensors offer opportunities for the development of new biosensors with much better performance. Biosensors were first introduced into the laboratory by Clark and Lyons. They developed the first glucose biosensor for laboratory conditions. Then in 1973, a glucose biosensor was commercialized by Yellow Springs Instruments. The commercial biosensors have small size and simple construction and they are ideal for point-of-care biosensing. In addition to glucose, a wide variety of metabolites such as lactate, cholesterol, and creatinine can be detected by using commercial biosensors. Like the glucose biosensors (tests) other commercial tests such as for pregnancy (hCG), Escherichia coli O157, influenza A and B viruses, Helicobacter pylori, human immunodeficiency virus, tuberculosis, and malaria have achieved success. Apart from their use in clinical analysis, commercial tests are also used in environmental (such as biochemical oxygen demand, nitrate, pesticide), food (such as glutamate, glutamine, sucrose, lactose, alcohol, ascorbic acid), and biothreat/biowarfare (Bacillus anthracis, Salmonella, Botulinum toxin) analysis. In this review, commercial biosensors in clinical, environmental, food, and biowarfare analysis are summarized and the commercial biosensors are compared in terms of their important characteristics. This is the first review in which all the commercially available tests are compiled together. Copyright © 2015 Elsevier Inc. All rights reserved.

The western transverse ranges microplate as a native terrane

DOE Office of Scientific and Technical Information (OSTI.GOV)

Campbell, M.D.; Reed, W.E.

1994-04-01

Palocurrent measurements from the entire Cretaceous section of the western Transverse Ranges microplate (WTRM) yield a northerly flow direction. Point count data indicate a mixed provenance for both conglomerates and associated sandstones. The dominant provenance was mixed magmatic arc/recycled orogen and disected/transitional arc terranes. Petrographic, quantitative SEM and microprobe analysis also indicate the presence of diagnostic Franciscan mineralogy in these sediments, including glaucophane, riebeckite, lawsonite, and serpentine, suggesting derivation from a subduction complex. Olistoclasts of chert, jadeitic graywacke, serpentine and blueschist are found intermixed within the arc-derived sediments. Olistoclasts range in size from sub-millimeter to centimeter scale and olistoliths rangemore » up to 150 m. Well preserved internal bedding in some of the olistoliths suggest emplacement by landsliding indicating very short transport distance. This Franciscan material represents the oldest melange-derived material reported from this part of California and documents uplift and erosion of the subduction complex earlier than previously suggested. These data are consistent with deposition in a Cretaceous fore-arc basin located west or south of the San Diego area. The allochthonous WTRM of southern California can be reconstructed to an originally north-south oriented fore-arc basin. After deposition of the Sespe Formation (22 Ma [+-]) the microplate was slivered by strike-slip faults and rotated clockwise approximately 90[degrees], after which, the block again accreted against the continental margin. Our reconstruction suggest that depositional and structural trends for Eocene and Cretaceous sediments is likely to be different from that in the Miocene Monterey pay zones in the Santa Barbara channel region. If our reconstruction is correct, exploration strategy for Eocene and Cretaceous petroleum in the southern California Bight should take this tectonic model into

Progress in chemical luminescence-based biosensors: A critical review.

PubMed

Roda, Aldo; Mirasoli, Mara; Michelini, Elisa; Di Fusco, Massimo; Zangheri, Martina; Cevenini, Luca; Roda, Barbara; Simoni, Patrizia

2016-02-15

Biosensors are a very active research field. They have the potential to lead to low-cost, rapid, sensitive, reproducible, and miniaturized bioanalytical devices, which exploit the high binding avidity and selectivity of biospecific binding molecules together with highly sensitive detection principles. Of the optical biosensors, those based on chemical luminescence detection (including chemiluminescence, bioluminescence, electrogenerated chemiluminescence, and thermochemiluminescence) are particularly attractive, due to their high-to-signal ratio and the simplicity of the required measurement equipment. Several biosensors based on chemical luminescence have been described for quantitative, and in some cases multiplex, analysis of organic molecules (such as hormones, drugs, pollutants), proteins, and nucleic acids. These exploit a variety of miniaturized analytical formats, such as microfluidics, microarrays, paper-based analytical devices, and whole-cell biosensors. Nevertheless, despite the high analytical performances described in the literature, the field of chemical luminescence biosensors has yet to demonstrate commercial success. This review presents the main recent advances in the field and discusses the approaches, challenges, and open issues, with the aim of stimulating a broader interest in developing chemical luminescence biosensors and improving their commercial exploitation. Copyright © 2015 Elsevier B.V. All rights reserved.

Analyzing the biosensor signal in flows: studies with glucose optrodes.

PubMed

Kivirand, K; Floren, A; Kagan, M; Avarmaa, T; Rinken, T; Jaaniso, R

2015-01-01

Responses of enzymatic bio-optrodes in flow regime were studied and an original model was proposed with the aim of establishing a reliable method for a quick determination of biosensor signal parameters, applicable for biosensor calibration. A dual-optrode glucose biosensor, comprising of a glucose bio-optrode and a reference oxygen optrode, both placed into identical flow channels, was developed and used as a model system. The signal parameters of this biosensor at different substrate concentrations were not dependent on the speed of the probe flow and could be determined from the initial part of the biosensor transient phase signal, providing a valuable tool for rapid analysis. In addition, the model helped to design the biosensor system with reduced impact of enzyme inactivation to the system stability (20% decrease of the enzyme activity lead to only a 1% decrease of the slope of the calibration curve) and hence significantly prolong the effective lifetime of bio-optrodes. Copyright © 2014 Elsevier B.V. All rights reserved.

A general strategy to construct small molecule biosensors in eukaryotes

DOE PAGES

Feng, Justin; Jester, Benjamin W.; Tinberg, Christine E.; ...

2015-12-29

Biosensors for small molecules can be used in applications that range from metabolic engineering to orthogonal control of transcription. Here, we produce biosensors based on a ligand-binding domain (LBD) by using a method that, in principle, can be applied to any target molecule. The LBD is fused to either a fluorescent protein or a transcriptional activator and is destabilized by mutation such that the fusion accumulates only in cells containing the target ligand. We illustrate the power of this method by developing biosensors for digoxin and progesterone. Addition of ligand to yeast, mammalian, or plant cells expressing a biosensor activatesmore » transcription with a dynamic range of up to ~100-fold. We use the biosensors to improve the biotransformation of pregnenolone to progesterone in yeast and to regulate CRISPR activity in mammalian cells. As a result, this work provides a general methodology to develop biosensors for a broad range of molecules in eukaryotes.« less

A general strategy to construct small molecule biosensors in eukaryotes

DOE Office of Scientific and Technical Information (OSTI.GOV)

Feng, Justin; Jester, Benjamin W.; Tinberg, Christine E.

Biosensors for small molecules can be used in applications that range from metabolic engineering to orthogonal control of transcription. Here, we produce biosensors based on a ligand-binding domain (LBD) by using a method that, in principle, can be applied to any target molecule. The LBD is fused to either a fluorescent protein or a transcriptional activator and is destabilized by mutation such that the fusion accumulates only in cells containing the target ligand. We illustrate the power of this method by developing biosensors for digoxin and progesterone. Addition of ligand to yeast, mammalian, or plant cells expressing a biosensor activatesmore » transcription with a dynamic range of up to ~100-fold. We use the biosensors to improve the biotransformation of pregnenolone to progesterone in yeast and to regulate CRISPR activity in mammalian cells. As a result, this work provides a general methodology to develop biosensors for a broad range of molecules in eukaryotes.« less

A general strategy to construct small molecule biosensors in eukaryotes

PubMed Central

Feng, Justin; Jester, Benjamin W; Tinberg, Christine E; Mandell, Daniel J; Antunes, Mauricio S; Chari, Raj; Morey, Kevin J; Rios, Xavier; Medford, June I; Church, George M; Fields, Stanley; Baker, David

2015-01-01

Biosensors for small molecules can be used in applications that range from metabolic engineering to orthogonal control of transcription. Here, we produce biosensors based on a ligand-binding domain (LBD) by using a method that, in principle, can be applied to any target molecule. The LBD is fused to either a fluorescent protein or a transcriptional activator and is destabilized by mutation such that the fusion accumulates only in cells containing the target ligand. We illustrate the power of this method by developing biosensors for digoxin and progesterone. Addition of ligand to yeast, mammalian, or plant cells expressing a biosensor activates transcription with a dynamic range of up to ~100-fold. We use the biosensors to improve the biotransformation of pregnenolone to progesterone in yeast and to regulate CRISPR activity in mammalian cells. This work provides a general methodology to develop biosensors for a broad range of molecules in eukaryotes. DOI: http://dx.doi.org/10.7554/eLife.10606.001 PMID:26714111

Portable guided-mode resonance biosensor platform for point-of-care testing

NASA Astrophysics Data System (ADS)

Sung, Gun Yong; Kim, Wan-Joong; Ko, Hyunsung; Kim, Bong K.; Kim, Kyung-Hyun; Huh, Chul; Hong, Jongcheol

2012-10-01

It represents a viable solution for the realization of a portable biosensor platform that could screen/diagnose acute myocardial infarction by measuring cardiac marker concentrations such as cardiac troponin I (cTnI), creatine kinase MB (CK-MB), and myoglobin (MYO) for application to u-health monitoring system. The portable biosensor platform introduced in this presentation has a more compact structure and a much higher measuring resolution than a conventional spectrometer system. Portable guided-mode resonance (GMR) biosensor platform was composed of a biosensor chip stage, an optical pick-up module, and a data display panel. Disposable plastic GMR biosensor chips with nano-grating patterns were fabricated by injection-molding. Whole blood filtration and label-free immunoassay were performed on these single chips, automatically. Optical pick-up module was fabricated by using the miniaturized bulk optics and the interconnecting optical fibers and a tunable VCSEL (vertical cavity surface emitting laser). The reflectance spectrum from the GMR biosensor was measured by the optical pick-up module. Cardiac markers in human serum with concentrations less than 0.1ng/mL were analyzed using a GMR biosensor. Analysis time was 30min, which is short enough to meet clinical requirements. Our results show that the GMR biosensor will be very useful in developing lowcost portable biosensors that can screen for cardiac diseases.

Array Biosensor for Toxin Detection: Continued Advances

PubMed Central

Taitt, Chris Rowe; Shriver-Lake, Lisa C.; Ngundi, Miriam M.; Ligler, Frances S.

2008-01-01

The following review focuses on progress made in the last five years with the NRL Array Biosensor, a portable instrument for rapid and simultaneous detection of multiple targets. Since 2003, the Array Biosensor has been automated and miniaturized for operation at the point-of-use. The Array Biosensor has also been used to demonstrate (1) quantitative immunoassays against an expanded number of toxins and toxin indicators in food and clinical fluids, and (2) the efficacy of semi-selective molecules as alternative recognition moieties. Blind trials, with unknown samples in a variety of matrices, have demonstrated the versatility, sensitivity, and reliability of the automated system. PMID:27873991

Biosensor-based engineering of biosynthetic pathways

DOE PAGES

Rogers, Jameson K.; Taylor, Noah D.; Church, George M.

2016-03-18

Biosynthetic pathways provide an enzymatic route from inexpensive renewable resources to valuable metabolic products such as pharmaceuticals and plastics. However, designing these pathways is challenging due to the complexities of biology. Advances in the design and construction of genetic variants has enabled billions of cells, each possessing a slightly different metabolic design, to be rapidly generated. However, our ability to measure the quality of these designs lags by several orders of magnitude. Recent research has enabled cells to report their own success in chemical production through the use of genetically encoded biosensors. A new engineering discipline is emerging around themore » creation and application of biosensors. Biosensors, implemented in selections and screens to identify productive cells, are paving the way for a new era of biotechnological progress.« less

Homemade Bienzymatic-Amperometric Biosensor for Beverages Analysis

ERIC Educational Resources Information Center

Blanco-Lopez, M. C.; Lobo-Castanon, M. J.; Miranda-Ordieres, A. J.

2007-01-01

The construction of an amperometric biosensor for glucose analysis is described demonstrating that the analysis is easy to perform and the biosensor gives good analytical performance. This experiment helped the students to acquire problem-solving and teamwork skills, allowing them to reach a high level of independent and critical thought.

Electrochemical Glucose Biosensor of Platinum Nanospheres Connected by Carbon Nanotubes

PubMed Central

Claussen, Jonathan C.; Kim, Sungwon S.; Haque, Aeraj ul; Artiles, Mayra S.; Porterfield, D. Marshall; Fisher, Timothy S.

2010-01-01

Background Glucose biosensors comprised of nanomaterials such as carbon nanotubes (CNTs) and metallic nanoparticles offer enhanced electrochemical performance that produces highly sensitive glucose sensing. This article presents a facile biosensor fabrication and biofunctionalization procedure that utilizes CNTs electrochemically decorated with platinum (Pt) nanospheres to sense glucose amperometrically with high sensitivity. Method Carbon nanotubes are grown in situ by microwave plasma chemical vapor deposition (MPCVD) and electro-chemically decorated with Pt nanospheres to form a CNT/Pt nanosphere composite biosensor. Carbon nanotube electrodes are immobilized with fluorescently labeled bovine serum albumin (BSA) and analyzed with fluorescence microscopy to demonstrate their biocompatibility. The enzyme glucose oxidase (GOX) is immobilized onto the CNT/Pt nanosphere biosensor by a simple drop-coat method for amperometric glucose sensing. Results Fluorescence microscopy demonstrates the biofunctionalization capability of the sensor by portraying adsorption of fluorescently labeled BSA unto MPCVD-grown CNT electrodes. The subsequent GOX–CNT/Pt nanosphere biosensor demonstrates a high sensitivity toward H2O2 (7.4 μA/mM/cm2) and glucose (70 μA/mM/cm2), with a glucose detection limit and response time of 380 nM (signal-to-noise ratio = 3) and 8 s (t90%), respectively. The apparent Michaelis–Menten constant (0.64 mM) of the biosensor also reflects the improved sensitivity of the immobilized GOX/nanomaterial complexes. Conclusions The GOX–CNT/Pt nanosphere biosensor outperforms similar CNT, metallic nanoparticle, and more conventional carbon-based biosensors in terms of glucose sensitivity and detection limit. The biosensor fabrication and biofunctionalization scheme can easily be scaled and adapted for microsensors for physiological research applications that require highly sensitive glucose sensing. PMID:20307391

Electrochemical glucose biosensor of platinum nanospheres connected by carbon nanotubes.

PubMed

Claussen, Jonathan C; Kim, Sungwon S; Haque, Aeraj Ul; Artiles, Mayra S; Porterfield, D Marshall; Fisher, Timothy S

2010-03-01

Glucose biosensors comprised of nanomaterials such as carbon nanotubes (CNTs) and metallic nanoparticles offer enhanced electrochemical performance that produces highly sensitive glucose sensing. This article presents a facile biosensor fabrication and biofunctionalization procedure that utilizes CNTs electrochemically decorated with platinum (Pt) nanospheres to sense glucose amperometrically with high sensitivity. Carbon nanotubes are grown in situ by microwave plasma chemical vapor deposition (MPCVD) and electro-chemically decorated with Pt nanospheres to form a CNT/Pt nanosphere composite biosensor. Carbon nanotube electrodes are immobilized with fluorescently labeled bovine serum albumin (BSA) and analyzed with fluorescence microscopy to demonstrate their biocompatibility. The enzyme glucose oxidase (GO(X)) is immobilized onto the CNT/Pt nanosphere biosensor by a simple drop-coat method for amperometric glucose sensing. Fluorescence microscopy demonstrates the biofunctionalization capability of the sensor by portraying adsorption of fluorescently labeled BSA unto MPCVD-grown CNT electrodes. The subsequent GO(X)-CNT/Pt nanosphere biosensor demonstrates a high sensitivity toward H(2)O(2) (7.4 microA/mM/cm(2)) and glucose (70 microA/mM/cm(2)), with a glucose detection limit and response time of 380 nM (signal-to-noise ratio = 3) and 8 s (t(90%)), respectively. The apparent Michaelis-Menten constant (0.64 mM) of the biosensor also reflects the improved sensitivity of the immobilized GO(X)/nanomaterial complexes. The GO(X)-CNT/Pt nanosphere biosensor outperforms similar CNT, metallic nanoparticle, and more conventional carbon-based biosensors in terms of glucose sensitivity and detection limit. The biosensor fabrication and biofunctionalization scheme can easily be scaled and adapted for microsensors for physiological research applications that require highly sensitive glucose sensing. (c) 2010 Diabetes Technology Society.

Progress in utilisation of graphene for electrochemical biosensors.

PubMed

Lawal, Abdulazeez T

2018-05-30

This review discusses recent graphene (GR) electrochemical biosensor for accurate detection of biomolecules, including glucose, hydrogen peroxide, dopamine, ascorbic acid, uric acid, nicotinamide adenine dinucleotide, DNA, metals and immunosensor through effective immobilization of enzymes, including glucose oxidase, horseradish peroxidase, and haemoglobin. GR-based biosensors exhibited remarkable performance with high sensitivities, wide linear detection ranges, low detection limits, and long-term stabilities. Future challenges for the field include miniaturising biosensors and simplifying mass production are discussed. Copyright © 2018 Elsevier B.V. All rights reserved.

Recent Advances in Application of Biosensors in Tissue Engineering

PubMed Central

Paul, Arghya; Lee, Yong-kyu; Jaffa, Ayad A.

2014-01-01

Biosensors research is a fast growing field in which tens of thousands of papers have been published over the years, and the industry is now worth billions of dollars. The biosensor products have found their applications in numerous industries including food and beverages, agricultural, environmental, medical diagnostics, and pharmaceutical industries and many more. Even though numerous biosensors have been developed for detection of proteins, peptides, enzymes, and numerous other biomolecules for diverse applications, their applications in tissue engineering have remained limited. In recent years, there has been a growing interest in application of novel biosensors in cell culture and tissue engineering, for example, real-time detection of small molecules such as glucose, lactose, and H2O2 as well as serum proteins of large molecular size, such as albumin and alpha-fetoprotein, and inflammatory cytokines, such as IFN-g and TNF-α. In this review, we provide an overview of the recent advancements in biosensors for tissue engineering applications. PMID:25165697

Recent advances in application of biosensors in tissue engineering.

PubMed

Hasan, Anwarul; Nurunnabi, Md; Morshed, Mahboob; Paul, Arghya; Polini, Alessandro; Kuila, Tapas; Al Hariri, Moustafa; Lee, Yong-kyu; Jaffa, Ayad A

2014-01-01

Biosensors research is a fast growing field in which tens of thousands of papers have been published over the years, and the industry is now worth billions of dollars. The biosensor products have found their applications in numerous industries including food and beverages, agricultural, environmental, medical diagnostics, and pharmaceutical industries and many more. Even though numerous biosensors have been developed for detection of proteins, peptides, enzymes, and numerous other biomolecules for diverse applications, their applications in tissue engineering have remained limited. In recent years, there has been a growing interest in application of novel biosensors in cell culture and tissue engineering, for example, real-time detection of small molecules such as glucose, lactose, and H2O2 as well as serum proteins of large molecular size, such as albumin and alpha-fetoprotein, and inflammatory cytokines, such as IFN-g and TNF-α. In this review, we provide an overview of the recent advancements in biosensors for tissue engineering applications.

Label-free optical detection of C-reactive protein by nanoimprint lithography-based 2D-photonic crystal film.

PubMed

Endo, Tatsuro; Kajita, Hiroshi; Kawaguchi, Yukio; Kosaka, Terumasa; Himi, Toshiyuki

2016-06-01

The development of high-sensitive, and cost-effective novel biosensors have been strongly desired for future medical diagnostics. To develop novel biosensor, the authors focused on the specific optical characteristics of photonic crystal. In this study, a label-free optical biosensor, polymer-based two-dimensional photonic crystal (2D-PhC) film fabricated using nanoimprint lithography (NIL), was developed for detection of C-reactive protein (CRP) in human serum. The nano-hole array constructed NIL-based 2D-PhC (hole diameter: 230 nm, distance: 230, depth: 200 nm) was fabricated on a cyclo-olefin polymer (COP) film (100 µm) using thermal NIL and required surface modifications to reduce nonspecific adsorption of target proteins. Antigen-antibody reactions on the NIL-based 2D-PhC caused changes to the surrounding refractive index, which was monitored as reflection spectrum changes in the visible region. By using surface modified 2D-PhC, the calculated detection limit for CRP was 12.24 pg/mL at an extremely short reaction time (5 min) without the need for additional labeling procedures and secondary antibody. Furthermore, using the dual-functional random copolymer, CRP could be detected in a pooled blood serum diluted 100× with dramatic reduction of nonspecific adsorption. From these results, the NIL-based 2D-PhC film has great potential for development of an on-site, high-sensitivity, cost-effective, label-free biosensor for medical diagnostics applications. © 2016 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

An electrochemiluminescence biosensor for endonuclease EcoRI detection.

PubMed

Li, Yingjie; Li, Yuqin; Wu, Yaoyu; Lu, Fushen; Chen, Yaowen; Gao, Wenhua

2017-03-15

Endonucleases cleavage of DNA plays an important role in biological and medicinal chemistry. This work was going to develop a reliable and sensitive electrochemiluminescent (ECL) biosensor for detecting endonucleases by using gold nanoparticles graphene composite (GNPs-graphene) as a signal amplifier. Firstly, the GNPs and graphene were simultaneously deposited on the glassy carbon electrode (GCE) by cyclic voltammetry. Then a stem DNA was anchored on the surface of GCE. And with a modifying DNA introduced into the electrode by DNA assembly, a strong ECL signal was obtained. After a DNA modified with ferrocene assembly to the stem DNA, the ECL signal had a sharp decrease due to the quench effect of ferrocene to and the biosensor comes into being a "off" state. With the effect of endonuclease, the ECL signal had a recovery because of the ferrocene being released and the biosensor formed a "on" state. Moreover, the recovery of ECL signal was related to the concentration of endonucleases. Combining specific defined DNA and endonuclease, this method has a potential to detect different endonucleases. In this work, we took the EcoRI as an example to identify the feasibility of ECL biosensor in applying in sensitive detection of endonucleases using a GNPs-graphene signal amplifier. Under optimal condition, the proposed biosensor obtained a low limit of detection (LOD) 5.6×10 -5 UmL -1 . And the stability, selectivity and reproducibility of the biosensor also were researched. Copyright © 2016 Elsevier B.V. All rights reserved.

Biosensor Regeneration: A Review of Common Techniques and Outcomes.

PubMed

Goode, J A; Rushworth, J V H; Millner, P A

2015-06-16

Biosensors are ideally portable, low-cost tools for the rapid detection of pathogens, proteins, and other analytes. The global biosensor market is currently worth over 10 billion dollars annually and is a burgeoning field of interdisciplinary research that is hailed as a potential revolution in consumer, healthcare, and industrial testing. A key barrier to the widespread adoption of biosensors, however, is their cost. Although many systems have been validated in the laboratory setting and biosensors for a range of analytes are proven at the concept level, many have yet to make a strong commercial case for their acceptance. Though it is true with the development of cheaper electrodes, circuits, and components that there is a downward pressure on costs, there is also an emerging trend toward the development of multianalyte biosensors that is pushing in the other direction. One way to reduce the cost that is suitable for certain systems is to enable their reuse, thus reducing the cost per test. Regenerating biosensors is a technique that can often be used in conjunction with existing systems in order to reduce costs and accelerate the commercialization process. This article discusses the merits and drawbacks of regeneration schemes that have been proven in various biosensor systems and indicates parameters for successful regeneration based on a systematic review of the literature. It also outlines some of the difficulties encountered when considering the role of regeneration at the point of use. A brief meta-analysis has been included in this review to develop a working definition for biosensor regeneration, and using this analysis only ∼60% of the reported studies analyzed were deemed a success. This highlights the variation within the field and the need to normalize regeneration as a standard process across the field by establishing a consensus term.

Two-Dimensional Photonic Crystals for Sensitive Microscale Chemical and Biochemical Sensing

PubMed Central

Miller, Benjamin L.

2015-01-01

Photonic crystals – optical devices able to respond to changes in the refractive index of a small volume of space – are an emerging class of label-free chemical-and bio-sensors. This review focuses on one class of photonic crystal, in which light is confined to a patterned planar material layer of sub-wavelength thickness. These devices are small (on the order of tens to 100s of microns square), suitable for incorporation into lab-on-a-chip systems, and in theory can provide exceptional sensitivity. We introduce the defining characteristics and basic operation of two-dimensional photonic crystal sensors, describe variations of their basic design geometry, and summarize reported detection results from chemical and biological sensing experiments. PMID:25563402

Development of human cell biosensor system for genotoxicity detection based on DNA damage-induced gene expression.

PubMed

Zager, Valerija; Cemazar, Maja; Hreljac, Irena; Lah, Tamara T; Sersa, Gregor; Filipic, Metka

2010-03-01

Human exposure to genotoxic agents in the environment and everyday life represents a serious health threat. Fast and reliable assessment of genotoxicity of chemicals is of main importance in the fields of new chemicals and drug development as well as in environmental monitoring. The tumor suppressor gene p21, the major downstream target gene of activated p53 which is responsible for cell cycle arrest following DNA damage, has been shown to be specifically up-regulated by genotoxic carcinogens. The aim of our study was to develop a human cell-based biosensor system for simple and fast detection of genotoxic agents. Metabolically active HepG2 human hepatoma cells were transfected with plasmid encoding Enhanced Green Fluorescent Protein (EGFP) under the control of the p21 promoter (p21HepG2GFP). DNA damage was induced by genotoxic agents with known mechanisms of action. The increase in fluorescence intensity, due to p21 mediated EGFP expression, was measured with a fluorescence microplate reader. The viability of treated cells was determined by the colorimetric MTS assay. The directly acting alkylating agent methylmethane sulphonate (MMS) showed significant increase in EGFP production after 48 h at 20 μg/mL. The indirectly acting carcinogen benzo(a)pyren (BaP) and the cross-linking agent cisplatin (CisPt) induced a dose- dependent increase in EGFP fluorescence, which was already significant at concentrations 0.13 μg/mL and 0.41 μg/mL, respectively. Vinblastine (VLB), a spindle poison that does not induce direct DNA damage, induced only a small increase in EGFP fluorescence intensity after 24 h at the lowest concentration (0.1 μg/mL), while exposure to higher concentrations was associated with significantly reduced cell viability. The results of our study demonstrated that this novel assay based on the stably transformed cell line p21HepG2GFP can be used as a fast and simple biosensor system for detection of genetic damage caused by chemical agents.

Future of biosensors: a personal view.

PubMed

Scheller, Frieder W; Yarman, Aysu; Bachmann, Till; Hirsch, Thomas; Kubick, Stefan; Renneberg, Reinhard; Schumacher, Soeren; Wollenberger, Ulla; Teller, Carsten; Bier, Frank F

2014-01-01

Biosensors representing the technological counterpart of living senses have found routine application in amperometric enzyme electrodes for decentralized blood glucose measurement, interaction analysis by surface plasmon resonance in drug development, and to some extent DNA chips for expression analysis and enzyme polymorphisms. These technologies have already reached a highly advanced level and need minor improvement at most. The dream of the "100-dollar" personal genome may come true in the next few years provided that the technological hurdles of nanopore technology or of polymerase-based single molecule sequencing can be overcome. Tailor-made recognition elements for biosensors including membrane-bound enzymes and receptors will be prepared by cell-free protein synthesis. As alternatives for biological recognition elements, molecularly imprinted polymers (MIPs) have been created. They have the potential to substitute antibodies in biosensors and biochips for the measurement of low-molecular-weight substances, proteins, viruses, and living cells. They are more stable than proteins and can be produced in large amounts by chemical synthesis. Integration of nanomaterials, especially of graphene, could lead to new miniaturized biosensors with high sensitivity and ultrafast response. In the future individual therapy will include genetic profiling of isoenzymes and polymorphic forms of drug-metabolizing enzymes especially of the cytochrome P450 family. For defining the pharmacokinetics including the clearance of a given genotype enzyme electrodes will be a useful tool. For decentralized online patient control or the integration into everyday "consumables" such as drinking water, foods, hygienic articles, clothing, or for control of air conditioners in buildings and cars and swimming pools, a new generation of "autonomous" biosensors will emerge.

Genetically engineered microbial biosensors for in situ monitoring of environmental pollution.

PubMed

Shin, Hae Ja

2011-02-01

Microbial biosensors are compact, portable, cost effective, and simple to use, making them seem eminently suitable for the in situ monitoring of environmental pollution. One promising approach for such applications is the fusion of reporter genes with regulatory genes that are dose-dependently responsive to the target chemicals or physiological signals. Their biosensor capabilities, such as target range and sensitivity, could be improved by modification of regulatory genes. Recent uses of such genetically engineered microbial biosensors include the development of portable biosensor kits and high-throughput cell arrays on chips, optic fibers, or other platforms for on-site and on-line monitoring of environmental pollution. This mini-review discusses recent advances in microbial biosensors and their future prospects, with a focus on the development and application of genetically modified microbial biosensors for in situ environmental monitoring.

Recent advances in electrochemical biosensors based on graphene two-dimensional nanomaterials.

PubMed

Song, Yang; Luo, Yanan; Zhu, Chengzhou; Li, He; Du, Dan; Lin, Yuehe

2016-02-15

Graphene as a star among two-dimensional nanomaterials has attracted tremendous research interest in the field of electrochemistry due to their intrinsic properties, including the electronic, optical, and mechanical properties associated with their planar structure. The marriage of graphene and electrochemical biosensors has created many ingenious biosensing strategies for applications in the areas of clinical diagnosis and food safety. This review provides a comprehensive overview of the recent advances in the development of graphene based electrochemical biosensors. Special attention is paid to graphene-based enzyme biosensors, immunosensors, and DNA biosensors. Future perspectives on high-performance graphene-based electrochemical biosensors are also discussed. Copyright © 2015 Elsevier B.V. All rights reserved.

Biosensors for Whole-Cell Bacterial Detection

PubMed Central

Rushworth, Jo V.; Hirst, Natalie A.; Millner, Paul A.

2014-01-01

SUMMARY Bacterial pathogens are important targets for detection and identification in medicine, food safety, public health, and security. Bacterial infection is a common cause of morbidity and mortality worldwide. In spite of the availability of antibiotics, these infections are often misdiagnosed or there is an unacceptable delay in diagnosis. Current methods of bacterial detection rely upon laboratory-based techniques such as cell culture, microscopic analysis, and biochemical assays. These procedures are time-consuming and costly and require specialist equipment and trained users. Portable stand-alone biosensors can facilitate rapid detection and diagnosis at the point of care. Biosensors will be particularly useful where a clear diagnosis informs treatment, in critical illness (e.g., meningitis) or to prevent further disease spread (e.g., in case of food-borne pathogens or sexually transmitted diseases). Detection of bacteria is also becoming increasingly important in antibioterrorism measures (e.g., anthrax detection). In this review, we discuss recent progress in the use of biosensors for the detection of whole bacterial cells for sensitive and earlier identification of bacteria without the need for sample processing. There is a particular focus on electrochemical biosensors, especially impedance-based systems, as these present key advantages in terms of ease of miniaturization, lack of reagents, sensitivity, and low cost. PMID:24982325

A novel conductometric biosensor based on hexokinase for determination of adenosine triphosphate.

PubMed

Kucherenko, I S; Kucherenko, D Yu; Soldatkin, O O; Lagarde, F; Dzyadevych, S V; Soldatkin, A P

2016-04-01

The paper presents a simple and inexpensive reusable biosensor for determination of the concentration of adenosine-5'-triphosphate (ATP) in aqueous samples. The biosensor is based on a conductometric transducer which contains two pairs of gold interdigitated electrodes. An enzyme hexokinase was immobilized onto one pair of electrodes, and bovine serum albumin-onto another pair (thus, a differential mode of measurement was used). Conditions of hexokinase immobilization on the transducer by cross-linking via glutaraldehyde were optimized. Influence of experimental conditions (concentration of magnesium ions, ionic strength and concentration of the working buffer) on the biosensor work was studied. The reproducibility of biosensor responses and operational stability of the biosensor were checked during one week. Dry storage at -18 °C was shown to be the best conditions to store the biosensor. The biosensor was successfully applied for measurements of ATP concentration in pharmaceutical samples. The proposed biosensor may be used in future for determination of ATP and/or glucose in water samples. Copyright © 2016 Elsevier B.V. All rights reserved.

Biosensors for marine applications. We all need the sea, but does the sea need biosensors?

PubMed

Kröger, Silke; Law, Robin J

2005-04-15

The aim of the paper is to explain the rationale behind marine biosensor applications, give an overview of measurement strategies currently employed, summarise some of the relevant available biosensor technology as well as instrumentation requirements for marine sensors and attempt a forward look at what the future might hold in terms of needs and developments. Application areas considered are eutrophication, organism detection, food safety, pollutants, trace metals and ecotoxicology. The drivers for many of these studies are discussed and the policy environment for current and future measurements is outlined.

Use of a parasitic wasp as a biosensor

USDA-ARS?s Scientific Manuscript database

Screening cargo for illicit substances is still in need of high-throughput inspection systems that can rapidly screen and accurately identify suspicious cargo. Here we investigate the ability of a parasitic wasp, Microplitis croceipes to detect and respond to methyl benzoate, the volatile component ...

High-Performance Multiplex SNP Analysis of Three Hemochromatosis-Related Mutations With Capillary Array Electrophoresis Microplates

PubMed Central

Medintz, Igor; Wong, Wendy W.; Berti, Lorenzo; Shiow, Lawrence; Tom, Jennifer; Scherer, James; Sensabaugh, George; Mathies, Richard A.

2001-01-01

An assay is described for high-throughput single nucleotide polymorphism (SNP) genotyping on a microfabricated capillary array electrophoresis (CAE) microchip. The assay targets the three common variants at the HFE locus associated with the genetic disease hereditary hemochromatosis (HHC). The assay employs allele-specific PCR (ASPCR) for the C282Y (845g->a), H63D (187c->g), and S65C (193a->t) variants using fluorescently-labeled energy-transfer (ET) allele-specific primers. Using a 96-channel radial CAE microplate, the labeled ASPCR products generated from 96 samples in a reference Caucasian population are simultaneously separated with single-base-pair resolution and genotyped in under 10 min. Detection is accomplished with a laser-excited rotary four-color fluorescence scanner. The allele-specific amplicons are differentiated on the basis of both their size and the color of the label emission. This study is the first demonstration of the combined use of ASPCR with ET primers and microfabricated radial CAE microplates to perform multiplex SNP analyses in a clinically relevant population. PMID:11230165

CMOS Electrochemical Instrumentation for Biosensor Microsystems: A Review.

PubMed

Li, Haitao; Liu, Xiaowen; Li, Lin; Mu, Xiaoyi; Genov, Roman; Mason, Andrew J

2016-12-31

Modern biosensors play a critical role in healthcare and have a quickly growing commercial market. Compared to traditional optical-based sensing, electrochemical biosensors are attractive due to superior performance in response time, cost, complexity and potential for miniaturization. To address the shortcomings of traditional benchtop electrochemical instruments, in recent years, many complementary metal oxide semiconductor (CMOS) instrumentation circuits have been reported for electrochemical biosensors. This paper provides a review and analysis of CMOS electrochemical instrumentation circuits. First, important concepts in electrochemical sensing are presented from an instrumentation point of view. Then, electrochemical instrumentation circuits are organized into functional classes, and reported CMOS circuits are reviewed and analyzed to illuminate design options and performance tradeoffs. Finally, recent trends and challenges toward on-CMOS sensor integration that could enable highly miniaturized electrochemical biosensor microsystems are discussed. The information in the paper can guide next generation electrochemical sensor design.

CMOS Electrochemical Instrumentation for Biosensor Microsystems: A Review

PubMed Central

Li, Haitao; Liu, Xiaowen; Li, Lin; Mu, Xiaoyi; Genov, Roman; Mason, Andrew J.

2016-01-01

Modern biosensors play a critical role in healthcare and have a quickly growing commercial market. Compared to traditional optical-based sensing, electrochemical biosensors are attractive due to superior performance in response time, cost, complexity and potential for miniaturization. To address the shortcomings of traditional benchtop electrochemical instruments, in recent years, many complementary metal oxide semiconductor (CMOS) instrumentation circuits have been reported for electrochemical biosensors. This paper provides a review and analysis of CMOS electrochemical instrumentation circuits. First, important concepts in electrochemical sensing are presented from an instrumentation point of view. Then, electrochemical instrumentation circuits are organized into functional classes, and reported CMOS circuits are reviewed and analyzed to illuminate design options and performance tradeoffs. Finally, recent trends and challenges toward on-CMOS sensor integration that could enable highly miniaturized electrochemical biosensor microsystems are discussed. The information in the paper can guide next generation electrochemical sensor design. PMID:28042860

Microplate and shear zone models for oceanic spreading center reorganizations

NASA Technical Reports Server (NTRS)

Engeln, Joseph F.; Stein, Seth; Werner, John; Gordon, Richard

1988-01-01

The kinematics of rift propagation and the resulting goemetries of various tectonic elements for two plates is reviewed with no overlap zone. The formation and evolution of overlap regions using schematic models is discussed. The models are scaled in space and time to approximate the Easter plate, but are simplified to emphasize key elements. The tectonic evolution of overlap regions which act as rigid microplates and shear zones is discussed, and the use of relative motion and structural data to discriminate between the two types of models is investigated. The effect of propagation rate and rise time on the size, shape, and deformation of the overlap region is demonstrated.

Enzyme-modified nanoporous gold-based electrochemical biosensors.

PubMed

Qiu, Huajun; Xue, Luyan; Ji, Guanglei; Zhou, Guiping; Huang, Xirong; Qu, Yinbo; Gao, Peiji

2009-06-15

On the basis of the unique physical and chemical properties of nanoporous gold (NPG), which was obtained simply by dealloying Ag from Au/Ag alloy, an attempt was made in the present study to develop NPG-based electrochemical biosensors. The NPG-modified glassy carbon electrode (NPG/GCE) exhibited high-electrocatalytic activity toward the oxidation of nicotinamide adenine dinucleotide (NADH) and hydrogen peroxide (H(2)O(2)), which resulted in a remarkable decrease in the overpotential of NADH and H(2)O(2) electro-oxidation when compared with the gold sheet electrode. The high density of edge-plane-like defective sites and large specific surface area of NPG should be responsible for the electrocatalytic behavior. Such electrocatalytic behavior of the NPG/GCE permitted effective low-potential amperometric biosensing of ethanol or glucose via the incorporation of alcohol dehydrogenase (ADH) or glucose oxidase (GOD) within the three-dimensional matrix of NPG. The ADH- and GOD-modified NPG-based biosensors showed good analytical performance for biosensing ethanol and glucose due to the clean, reproducible and uniformly distributed microstructure of NPG. The stabilization effect of NPG on the incorporated enzymes also made the constructed biosensors very stable. After 1 month storage at 4 degrees C, the ADH- and GOD-based biosensors lost only 5.0% and 4.2% of the original current response. All these indicated that NPG was a promising electrode material for biosensors construction.

Glucose Biosensors: An Overview of Use in Clinical Practice

PubMed Central

Yoo, Eun-Hyung; Lee, Soo-Youn

2010-01-01

Blood glucose monitoring has been established as a valuable tool in the management of diabetes. Since maintaining normal blood glucose levels is recommended, a series of suitable glucose biosensors have been developed. During the last 50 years, glucose biosensor technology including point-of-care devices, continuous glucose monitoring systems and noninvasive glucose monitoring systems has been significantly improved. However, there continues to be several challenges related to the achievement of accurate and reliable glucose monitoring. Further technical improvements in glucose biosensors, standardization of the analytical goals for their performance, and continuously assessing and training lay users are required. This article reviews the brief history, basic principles, analytical performance, and the present status of glucose biosensors in the clinical practice. PMID:22399892

Interdigitated electrodes as impedance and capacitance biosensors: A review

NASA Astrophysics Data System (ADS)

Mazlan, N. S.; Ramli, M. M.; Abdullah, M. M. A. B.; Halin, D. S. C.; Isa, S. S. M.; Talip, L. F. A.; Danial, N. S.; Murad, S. A. Z.

2017-09-01

Interdigitated electrodes (IDEs) are made of two individually addressable interdigitated comb-like electrode structures. IDEs are one of the most favored transducers, widely utilized in technological applications especially in the field of biological and chemical sensors due to their inexpensive, ease of fabrication process and high sensitivity. In order to detect and analyze a biochemical molecule or analyte, the impedance and capacitance signal need to be obtained. This paper investigates the working principle and influencer of the impedance and capacitance biosensors. The impedance biosensor depends on the resistance and capacitance while the capacitance biosensor influenced by the dielectric permittivity. However, the geometry and structures of the interdigitated electrodes affect both impedance and capacitance biosensor. The details have been discussed in this paper.

Film bulk acoustic resonators (FBARs) as biosensors: A review.

PubMed

Zhang, Yi; Luo, Jikui; Flewitt, Andrew J; Cai, Zhiqiang; Zhao, Xiubo

2018-09-30

Biosensors play important roles in different applications such as medical diagnostics, environmental monitoring, food safety, and the study of biomolecular interactions. Highly sensitive, label-free and disposable biosensors are particularly desired for many clinical applications. In the past decade, film bulk acoustic resonators (FBARs) have been developed as biosensors because of their high resonant frequency and small base mass (hence greater sensitivity), lower cost, label-free capability and small size. This paper reviews the piezoelectric materials used for FBARs, the optimisation of device structures, and their applications as biosensors in a wide range of biological applications such as the detection of antigens, DNAs and small biomolecules. Their integration with microfluidic devices and high-throughput detection are also discussed. Copyright © 2018 Elsevier B.V. All rights reserved.

Acetylcholinesterase-reduced graphene oxide hybrid films for organophosphorus neurotoxin sensing via quartz crystal microbalance

NASA Astrophysics Data System (ADS)

Tang, Shi; Ma, Wenying; Xie, Guangzhong; Su, Yuanjie; Jiang, Yadong

2016-09-01

An acetylcholinesterase (AChE)-reduced graphene oxide (RGO) hybrid films based biosensor enabled by quartz crystal microbalance (QCM) has been developed for the detection of organophosphorus neurotoxin in gas phase at room temperature. To improve the sensing performance, RGO was used to immobilize large quantities of enzyme and provide a favorable microenvironment to maintain the enzyme activity. The experimental results reveal that the response of AChE-RGO/glutaraldehyde based sensors is about 8 times larger than that of the AChE with the sensitivity of 1.583 Hz/mg/m3. 1.0 mg amount of RGO, 5% concentration of glutaraldehyde and pH 6.8 is the optimal condition of this biosensor.

Investigation of ultrahigh sensitivity in GaInAsP nanolaser biosensor

NASA Astrophysics Data System (ADS)

Saijo, Yoshito; Watanabe, Takumi; Hasegawa, Yu; Nishijima, Yoshiaki; Baba, Toshihiko

2018-02-01

We have developed GaInAsP semiconductor photonic crystal nanolaser biosensor and demonstrated the detection of ultralow-concentration (fM to aM) proteins and deoxyribonucleic acids (DNAs) adsorbed on the device surface. In general, this type of photonic sensors exploiting optical resonance has been considered to detect the refractive index of biomolecules via the wavelength shift. However, this principle cannot explain the detection of such ultralowconcentration. Therefore, we investigated another candidate principle, i.e., ion sensitivity. We consider such a process that 1) the electric charge of biomolecules changes the nanolaser's surface charge, 2) the Schottky barrier near the semiconductor surface is increased or decreased, 3) the distribution of photopumped carriers is modified by the barrier, 4) the refractive index of the semiconductor is changed by the carrier effects, and 5) the laser wavelength shifts. To confirm this process, we electrochemically measured the zeta and flatband potentials when charged electrolyte polymers were adsorbed in water. We clearly observed that these potentials temporally behaved consistently with that of the laser wavelength, which suggests that polymers significantly acted on the Schottky barrier. The same behaviors were also observed for the adsorption of 1 fM DNA. We consider that a limited number of charged DNA changed the surface functional group of the entire device surface. Such charge effects will be the key that achieves the ultrahigh sensitivity in the nanolaser biosensor.

Engineering nanomaterials-based biosensors for food safety detection.

PubMed

Lv, Man; Liu, Yang; Geng, Jinhui; Kou, Xiaohong; Xin, Zhihong; Yang, Dayong

2018-05-30

Food safety always remains a grand global challenge to human health, especially in developing countries. To solve food safety pertained problems, numerous strategies have been developed to detect biological and chemical contaminants in food. Among these approaches, nanomaterials-based biosensors provide opportunity to realize rapid, sensitive, efficient and portable detection, overcoming the restrictions and limitations of traditional methods such as complicated sample pretreatment, long detection time, and relying on expensive instruments and well-trained personnel. In this review article, we provide a cross-disciplinary perspective to review the progress of nanomaterials-based biosensors for the detection of food contaminants. The review article is organized by the category of food contaminants including pathogens/toxins, heavy metals, pesticides, veterinary drugs and illegal additives. In each category of food contaminant, the biosensing strategies are summarized including optical, colorimetric, fluorescent, electrochemical, and immune- biosensors; the relevant analytes, nanomaterials and biosensors are analyzed comprehensively. Future perspectives and challenges are also discussed briefly. We envision that our review could bridge the gap between the fields of food science and nanotechnology, providing implications for the scientists or engineers in both areas to collaborate and promote the development of nanomaterials-based biosensors for food safety detection. Copyright © 2018 Elsevier B.V. All rights reserved.

Flexible Molybdenum Electrodes towards Designing Affinity Based Protein Biosensors

PubMed Central

Kamakoti, Vikramshankar; Panneer Selvam, Anjan; Radha Shanmugam, Nandhinee; Muthukumar, Sriram; Prasad, Shalini

2016-01-01

Molybdenum electrode based flexible biosensor on porous polyamide substrates has been fabricated and tested for its functionality as a protein affinity based biosensor. The biosensor performance was evaluated using a key cardiac biomarker; cardiac Troponin-I (cTnI). Molybdenum is a transition metal and demonstrates electrochemical behavior upon interaction with an electrolyte. We have leveraged this property of molybdenum for designing an affinity based biosensor using electrochemical impedance spectroscopy. We have evaluated the feasibility of detection of cTnI in phosphate-buffered saline (PBS) and human serum (HS) by measuring impedance changes over a frequency window from 100 mHz to 1 MHz. Increasing changes to the measured impedance was correlated to the increased dose of cTnI molecules binding to the cTnI antibody functionalized molybdenum surface. We achieved cTnI detection limit of 10 pg/mL in PBS and 1 ng/mL in HS medium. The use of flexible substrates for designing the biosensor demonstrates promise for integration with a large-scale batch manufacturing process. PMID:27438863

Flexible Molybdenum Electrodes towards Designing Affinity Based Protein Biosensors.

PubMed

Kamakoti, Vikramshankar; Panneer Selvam, Anjan; Radha Shanmugam, Nandhinee; Muthukumar, Sriram; Prasad, Shalini

2016-07-18

Molybdenum electrode based flexible biosensor on porous polyamide substrates has been fabricated and tested for its functionality as a protein affinity based biosensor. The biosensor performance was evaluated using a key cardiac biomarker; cardiac Troponin-I (cTnI). Molybdenum is a transition metal and demonstrates electrochemical behavior upon interaction with an electrolyte. We have leveraged this property of molybdenum for designing an affinity based biosensor using electrochemical impedance spectroscopy. We have evaluated the feasibility of detection of cTnI in phosphate-buffered saline (PBS) and human serum (HS) by measuring impedance changes over a frequency window from 100 mHz to 1 MHz. Increasing changes to the measured impedance was correlated to the increased dose of cTnI molecules binding to the cTnI antibody functionalized molybdenum surface. We achieved cTnI detection limit of 10 pg/mL in PBS and 1 ng/mL in HS medium. The use of flexible substrates for designing the biosensor demonstrates promise for integration with a large-scale batch manufacturing process.

Recent advances in transition-metal dichalcogenides based electrochemical biosensors: A review.

PubMed

Wang, Yi-Han; Huang, Ke-Jing; Wu, Xu

2017-11-15

Layered transition metal dichalcogenides (TMDCs) comprise a category of two-dimensional (2D) materials that offer exciting properties, including large surface area, metallic and semi-conducting electrical capabilities, and intercalatable morphologies. Biosensors employ biological molecules to recognize the target and utilize output elements which can translate the biorecognition event into electrical, optical or mass-sensitive signals to determine the quantities of the target. TMDCs nanomaterials have been widely applied in various electrochemical biosensors with high sensitivity and selectivity. The marriage of TMDCs and electrochemical biosensors has created many productive sensing strategies for applications in the areas of clinical diagnosis, environmental monitoring and food safety. In recent years, an increasing number of TMDCs-based electrochemical biosensors are reported, suggesting TMDCs offers new possibilities of improving the performance of electrochemical biosensors. This review summarizes recent advances in electrochemical biosensors based on TMDCs for detection of various inorganic and organic analytes in the last five years, including glucose, proteins, DNA, heavy metal, etc. In addition, we also point out the challenges and future perspectives related to the material design and development of TMDCs-based electrochemical biosensors. Copyright © 2017 Elsevier B.V. All rights reserved.

A Microplate Growth Inhibition Assay for Screening Bacteriocins against Listeria monocytogenes to Differentiate Their Mode-of-Action.

PubMed

Vijayakumar, Paul Priyesh; Muriana, Peter M

2015-06-11

Lactic acid bacteria (LAB) have historically been used in food fermentations to preserve foods and are generally-recognized-as-safe (GRAS) by the FDA for use as food ingredients. In addition to lactic acid; some strains also produce bacteriocins that have been proposed for use as food preservatives. In this study we examined the inhibition of Listeria monocytogenes 39-2 by neutralized and non-neutralized bacteriocin preparations (Bac+ preps) produced by Lactobacillus curvatus FS47; Lb. curvatus Beef3; Pediococcus acidilactici Bac3; Lactococcus lactis FLS1; Enterococcus faecium FS56-1; and Enterococcus thailandicus FS92. Activity differences between non-neutralized and neutralized Bac+ preps in agar spot assays could not readily be attributed to acid because a bacteriocin-negative control strain was not inhibitory to Listeria in these assays. When neutralized and non-neutralized Bac+ preps were used in microplate growth inhibition assays against L. monocytogenes 39-2 we observed some differences attributed to acid inhibition. A microplate growth inhibition assay was used to compare inhibitory reactions of wild-type and bacteriocin-resistant variants of L. monocytogenes to differentiate bacteriocins with different modes-of-action (MOA) whereby curvaticins FS47 and Beef3, and pediocin Bac3 were categorized to be in MOA1; enterocins FS92 and FS56-1 in MOA2; and lacticin FLS1 in MOA3. The microplate bacteriocin MOA assay establishes a platform to evaluate the best combination of bacteriocin preparations for use in food applications as biopreservatives against L. monocytogenes.

A Microplate Growth Inhibition Assay for Screening Bacteriocins against Listeria monocytogenes to Differentiate Their Mode-of-Action

PubMed Central

Vijayakumar, Paul Priyesh; Muriana, Peter M.

2015-01-01

Lactic acid bacteria (LAB) have historically been used in food fermentations to preserve foods and are generally-recognized-as-safe (GRAS) by the FDA for use as food ingredients. In addition to lactic acid; some strains also produce bacteriocins that have been proposed for use as food preservatives. In this study we examined the inhibition of Listeria monocytogenes 39-2 by neutralized and non-neutralized bacteriocin preparations (Bac+ preps) produced by Lactobacillus curvatus FS47; Lb. curvatus Beef3; Pediococcus acidilactici Bac3; Lactococcus lactis FLS1; Enterococcus faecium FS56-1; and Enterococcus thailandicus FS92. Activity differences between non-neutralized and neutralized Bac+ preps in agar spot assays could not readily be attributed to acid because a bacteriocin-negative control strain was not inhibitory to Listeria in these assays. When neutralized and non-neutralized Bac+ preps were used in microplate growth inhibition assays against L. monocytogenes 39-2 we observed some differences attributed to acid inhibition. A microplate growth inhibition assay was used to compare inhibitory reactions of wild-type and bacteriocin-resistant variants of L. monocytogenes to differentiate bacteriocins with different modes-of-action (MOA) whereby curvaticins FS47 and Beef3, and pediocin Bac3 were categorized to be in MOA1; enterocins FS92 and FS56-1 in MOA2; and lacticin FLS1 in MOA3. The microplate bacteriocin MOA assay establishes a platform to evaluate the best combination of bacteriocin preparations for use in food applications as biopreservatives against L. monocytogenes. PMID:26111195

Population subdivision of hydrothermal vent polychaete Alvinella pompejana across equatorial and Easter Microplate boundaries.

PubMed

Jang, Sook-Jin; Park, Eunji; Lee, Won-Kyung; Johnson, Shannon B; Vrijenhoek, Robert C; Won, Yong-Jin

2016-10-28

The Equator and Easter Microplate regions of the eastern Pacific Ocean exhibit geomorphological and hydrological features that create barriers to dispersal for a number of animals associated with deep-sea hydrothermal vent habitats. This study examined effects of these boundaries on geographical subdivision of the vent polychaete Alvinella pompejana. DNA sequences from one mitochondrial and eleven nuclear genes were examined in samples collected from ten vent localities that comprise the species' known range from 23°N latitude on the East Pacific Rise to 38°S latitude on the Pacific Antarctic Ridge. Multi-locus genotypes inferred from these sequences clustered the individual worms into three metapopulation segments - the northern East Pacific Rise (NEPR), southern East Pacific Rise (SEPR), and northeastern Pacific Antarctic Ridge (PAR) - separated by the Equator and Easter Microplate boundaries. Genetic diversity estimators were negatively correlated with tectonic spreading rates. Application of the isolation-with-migration (IMa2) model provided information about divergence times and demographic parameters. The PAR and NEPR metapopulation segments were estimated to have split roughly 4.20 million years ago (Mya) (2.42-33.42 Mya, 95 % highest posterior density, (HPD)), followed by splitting of the SEPR and NEPR segments about 0.79 Mya (0.07-6.67 Mya, 95 % HPD). Estimates of gene flow between the neighboring regions were mostly low (2 Nm < 1). Estimates of effective population size decreased with southern latitudes: NEPR > SEPR > PAR. Highly effective dispersal capabilities allow A. pompejana to overcome the temporal instability and intermittent distribution of active hydrothermal vents in the eastern Pacific Ocean. Consequently, the species exhibits very high levels of genetic diversity compared with many co-distributed vent annelids and mollusks. Nonetheless, its levels of genetic diversity in partially isolated populations are inversely

Characterization of growth inhibition of oral bacteria by sophorolipid using a microplate-format assay.

PubMed

Solaiman, Daniel K Y; Ashby, Richard D; Uknalis, Joseph

2017-05-01

Sophorolipid (SL) is a class of glycolipid biosurfactant produced by yeast and has potent antimicrobial activity against many microorganisms. In this paper, a microplate-based method was developed to characterize the growth inhibition by SL on five representative species of caries-causing oral bacteria. Bacterial growth on microplate in the absence and presence of varying concentrations of SL was continuously monitored by recording the absorbance at 600nm of the cultures using a microplate reader. The results showed that SL completely inhibited the growth of the Lactobacilli at ≥1mg/ml and the Streptococci at much lower concentrations of ≥50μg/ml. More importantly, we further defined the mechanism of antimicrobial activity of SL by analyzing the pattern of the cell growth curves. SL at sublethal concentrations (<1mg/ml) is bactericidal towards the Lactobacilli; it lengthens the apparent cell-doubling time (T d ) and decreases the final cell density (as indicated by A 600nm ) in a concentration-dependent manner. Against the oral Streptococci, on the other hand, SL at sublethal concentrations (<50μg/ml) is bacteriostatic; it delays the onset of cell growth in a concentration-dependent fashion, but once the cell growth is commenced there is no noticeable adverse effect on T d and the final A 600nm . Scanning electron microscopic (SEM) study of L. acidophilus grown in sublethal concentration of SL reveals extensive structural damage to the cells. S. mutans grown in sublethal level of SL did not show morphological damage to the cells, but numerous protruding structures could be seen on the cell surface. At the respective lethal levels of SL, L. acidophilus cells were lysed (at 1mg/ml SL) and the cell surface structure of S. mutans (at 130μg/ml SL) was extensively deformed. In summary, this paper presents the first report on a detailed analysis of the effects of SL on Lactobacilli and Streptococci important to oral health and hygiene. Published by Elsevier B.V.

Nanoplasmonic Biosensor Using Localized Surface Plasmon Resonance Spectroscopy for Biochemical Detection.

PubMed

Zhang, Diming; Zhang, Qian; Lu, Yanli; Yao, Yao; Li, Shuang; Liu, Qingjun

2017-01-01

Localized surface plasmon resonance (LSPR) associated with metal nanostructures has developed into a highly useful sensor technique. Optical LSPR spectroscopy of nanostructures often shows sharp absorption and scattering peaks, which can be used to probe several bio-molecular interactions. Here, we report nanoplasmonic biosensors using LSPR on nanocup arrays (nanoCA) to recognize bio-molecular binding for biochemical detection. These sensors can be modified to quantify binding of small molecules to proteins for odorant and explosive detections. Electrochemical LSPR biosensors can also be designed by coupling electrochemistry and LSPR spectroscopy measurements. Multiple sensing information can be obtained and electrochemical LSPR property can be investigated for biosensors. In some applications, the electrochemical LSPR biosensor can be used to quantify immunoreactions and enzymatic activity. The biosensors exhibit better performance than those of conventional optical LSPR measurements. With multi-transducers, the nanoplasmonic biosensor can provide a promising approach for bio-detection in environmental monitoring, healthcare diagnostics, and food quality control.

Amperometric biosensor system for simultaneous determination of adenosine-5'-triphosphate and glucose.

PubMed

Kucherenko, Ivan S; Didukh, Daria Yu; Soldatkin, Oleksandr O; Soldatkin, Alexei P

2014-06-03

The majority of biosensors for adenosine-5'-triphosphate (ATP) determination are based on cascades of enzymatic reactions; therefore, they are sensitive to glucose or glycerol (depending on the enzymatic system) as well as to ATP. The presence of unknown concentrations of these substances in the sample greatly complicates the determination of ATP. To overcome this disadvantage of known biosensors, we developed a biosensor system consisting of two biosensors: the first one is based on glucose oxidase and is intended for measuring glucose concentration, and the second one is based on glucose oxidase and hexokinase and is sensitive toward both glucose and ATP. Using glucose concentration measured by the first biosensor, we can analyze the total response to glucose and ATP obtained by the second biosensor. Platinum disc electrodes were used as amperometric transducers. The polyphenilenediamine membrane was deposited onto the surface of platinum electrodes to avoid the response to electroactive substances. The effect of glucose concentration on biosensor determination of ATP was studied. The reproducibility of biosensor responses to glucose and ATP during a day was tested (relative standard deviation, RSD, of responses to glucose was 3-6% and to ATP was 8-12%) as well as storage stability of the biosensors (no decrease of glucose responses and 43% drop of ATP responses during 50 days). The measurements of ATP and glucose in pharmaceutical vials (including mixtures of ATP and glucose) were carried out. It was shown that the developed biosensor system can be used for simultaneous analysis of glucose and ATP concentrations in water solutions.

Mercaptobenzothiazole-on-gold organic phase biosensor systems: 1. Enhanced organosphosphate pesticide determination.

PubMed

Somerset, V; Baker, P; Iwuoha, E

2009-02-01

This paper reports the construction of the gold/mercaptobenzothiazole/polyaniline/acetylcholinesterase/polyvinylacetate (Au/ MBT/PANI/AChE/PVAc) thick-film biosensor for the determination of certain organophosphate pesticide solutions in selected aqueous organic solvent solutions. The Au/MBT/PANI/AChE/PVAc electrocatalytic biosensor device was constructed by encapsulating acetylcholinesterase (AChE) enzyme in the PANI polymer composite, followed by the coating of poly(vinyl acetate) (PVAc) on top to secure the biosensor film from disintegration in the organic solvents evaluated. The electroactive substrate called acetylthiocholine (ATCh) was employed to provide the movement of electrons in the amperometric biosensor. The voltammetric results have shown that the current shifts more anodically as the Au/MBT/PANI/AChE/PVAc biosensor responded to successive acetylthiocholine (ATCh) substrate addition under anaerobic conditions in 0.1 M phosphate buffer, KCl (pH 7.2) solution and aqueous organic solvent solutions. For the Au/MBT/PANI/AChE/PVAc biosensor, various performance and stability parameters were evaluated. These factors include the optimal enzyme loading, effect of pH, long-term stability of the biosensor, temperature stability of the biosensor, the effect of polar organic solvents, and the effect of non-polar organic solvents on the amperometric behavior of the biosensor. The biosensor was then applied to detect a series of 5 organophosphorous pesticides in aqueous organic solvents and the pesticides studied were parathion-methyl, malathion and chlorpyrifos. The results obtained have shown that the detection limit values for the individual pesticides were 1.332 nM (parathion-methyl), 0.189 nM (malathion), 0.018 nM (chlorpyrifos).

Lignin and silicate based hydrogels for biosensor applications

NASA Astrophysics Data System (ADS)

Burrs, S. L.; Jairam, S.; Vanegas, D. C.; Tong, Z.; McLamore, E. S.

2013-05-01

Advances in biocompatible materials and electrocatalytic nanomaterials have extended and enhanced the field of biosensors. Immobilization of biorecognition elements on nanomaterial platforms is an efficient technique for developing high fidelity biosensors. Single layer (i.e., Langmuir-Blodgett) protein films are efficient, but disadvantages of this approach include high cost, mass transfer limitations, and Vromer competition for surface binding sites. There is a need for simple, user friendly protein-nanomaterial sensing membranes that can be developed in laboratories or classrooms (i.e., outside of the clean room). In this research, we develop high fidelity nanomaterial platforms for developing electrochemical biosensors using sustainable biomaterials and user-friendly deposition techniques. Catalytic nanomaterial platforms are developed using a combination of self assembled monolayer chemistry and electrodeposition. High performance biomaterials (e.g., nanolignin) are recovered from paper pulp waste and combined with proteins and nanomaterials to form active sensor membranes. These methods are being used to develop electrochemical biosensors for studying physiological transport in biomedical, agricultural, and environmental applications.

Resonance phenomenon of the ATP motor as an ultrasensitive biosensor.

PubMed

Wang, Peirong; Zhang, Xiaoguang; Zhang, Xu; Wang, Xia; Li, Xueren; Yue, Jiachang

2012-09-28

We designed a rotary biosensor as a damping effector, with the rotation of the F(0)F(1)-ATPase driven by Adenosine Triphosphate (ATP) synthesis being indicated by the fluorescence intensity and a damping effect force being induced by the binding of an RNA molecule to its probe on the rotary biosensor. We found that the damping effect could contribute to the resonance phenomenon and energy transfer process of our rotary biosensor in the liquid phase. This result indicates that the ability of the rotary motor to operate in the vibration harmonic mode depends on the environmental conditions and mechanism in that a few molecules of the rotary biosensor could induce all of the sensor molecules to fluoresce together. These findings contribute to the theory study of the ATPase motor and future development of biosensors for ultrasensitive detection. Copyright © 2012 Elsevier Inc. All rights reserved.

Sensitive microplate assay for the detection of proteolytic enzymes using radiolabeled gelatin

DOE Office of Scientific and Technical Information (OSTI.GOV)

Robertson, B.D.; Kwan-Lim, G.E.; Maizels, R.M.

1988-07-01

A sensitive, microplate assay is described for the detection of a wide range of proteolytic enzymes, using radio-iodine-labeled gelatin as substrate. The technique uses the Bolton-Hunter reagent to label the substrate, which is then coated onto the wells of polyvinyl chloride microtiter plates. By measuring the radioactivity released the assay is able to detect elastase, trypsin, and collagenase in concentrations of 1 ng/ml or less, while the microtiter format permits multiple sample handling and minimizes sample volumes required for analysis.

Development and Applications of Portable Biosensors.

PubMed

Srinivasan, Balaji; Tung, Steve

2015-08-01

The significance of microfluidics-based and microelectromechanical systems-based biosensors has been widely acknowledged, and many reviews have explored their potential applications in clinical diagnostics, personalized medicine, global health, drug discovery, food safety, and forensics. Because health care costs are increasing, there is an increasing need to remotely monitor the health condition of patients by point-of-care-testing. The demand for biosensors for detection of biological warfare agents has increased, and research is focused on ways of producing small portable devices that would allow fast, accurate, and on-site detection. In the past decade, the demand for rapid and accurate on-site detection of plant disease diagnosis has increased due to emerging pathogens with resistance to pesticides, increased human mobility, and regulations limiting the application of toxic chemicals to prevent spread of diseases. The portability of biosensors for on-site diagnosis is limited due to various issues, including sample preparation techniques, fluid-handling techniques, the limited lifetime of biological reagents, device packaging, integrating electronics for data collection/analysis, and the requirement of external accessories and power. Many microfluidic, electronic, and biological design strategies, such as handling liquids in biosensors without pumps/valves, the application of droplet-based microfluidics, paper-based microfluidic devices, and wireless networking capabilities for data transmission, are being explored. © 2015 Society for Laboratory Automation and Screening.

Scattering-Type Surface-Plasmon-Resonance Biosensors

NASA Technical Reports Server (NTRS)

Wang, Yu; Pain, Bedabrata; Cunningham, Thomas; Seshadri, Suresh

2005-01-01

Biosensors of a proposed type would exploit scattering of light by surface plasmon resonance (SPR). Related prior biosensors exploit absorption of light by SPR. Relative to the prior SPR biosensors, the proposed SPR biosensors would offer greater sensitivity in some cases, enough sensitivity to detect bioparticles having dimensions as small as nanometers. A surface plasmon wave can be described as a light-induced collective oscillation in electron density at the interface between a metal and a dielectric. At SPR, most incident photons are either absorbed or scattered at the metal/dielectric interface and, consequently, reflected light is greatly attenuated. The resonance wavelength and angle of incidence depend upon the permittivities of the metal and dielectric. An SPR sensor of the type most widely used heretofore includes a gold film coated with a ligand a substance that binds analyte molecules. The gold film is thin enough to support evanescent-wave coupling through its thickness. The change in the effective index of refraction at the surface, and thus the change in the SPR response, increases with the number of bound analyte molecules. The device is illuminated at a fixed wavelength, and the intensity of light reflected from the gold surface opposite the ligand-coated surface is measured as a function of the angle of incidence. From these measurements, the angle of minimum reflection intensity is determined

Nanomaterials-based enzyme electrochemical biosensors operating through inhibition for biosensing applications.

PubMed

Kurbanoglu, Sevinc; Ozkan, Sibel A; Merkoçi, Arben

2017-03-15

In recent years great progress has been made in applying nanomaterials to design novel biosensors. Use of nanomaterials offers to biosensing platforms exceptional optical, electronic and magnetic properties. Nanomaterials can increase the surface of the transducing area of the sensors that in turn bring an increase in catalytic behaviors. They have large surface-to-volume ratio, controlled morphology and structure that also favor miniaturization, an interesting advantage when the sample volume is a critical issue. Biosensors have great potential for achieving detect-to-protect devices: devices that can be used in detections of pollutants and other treating compounds/analytes (drugs) protecting citizens' life. After a long term focused scientific and financial efforts/supports biosensors are expected now to fulfill their promise such as being able to perform sampling and analysis of complex samples with interest for clinical or environment fields. Among all types of biosensors, enzymatic biosensors, the most explored biosensing devices, have an interesting property, the inherent inhibition phenomena given the enzyme-substrate complex formation. The exploration of such phenomena is making remarkably important their application as research and applied tools in diagnostics. Different inhibition biosensor systems based on nanomaterials modification has been proposed and applied. The role of nanomaterials in inhibition-based biosensors for the analyses of different groups of drugs as well as contaminants such as pesticides, phenolic compounds and others, are discussed in this review. This deep analysis of inhibition-based biosensors that employ nanomaterials will serve researchers as a guideline for further improvements and approaching of these devices to real sample applications so as to reach society needs and such biosensor market demands. Copyright © 2016 Elsevier B.V. All rights reserved.

Sensors and Biosensors for C-Reactive Protein, Temperature and pH, and Their Applications for Monitoring Wound Healing: A Review

PubMed Central

Dini, Valentina; Kirchhain, Arno; Janowska, Agata; Oranges, Teresa; Di Francesco, Fabio

2017-01-01

Wound assessment is usually performed in hospitals or specialized labs. However, since patients spend most of their time at home, a remote real time wound monitoring would help providing a better care and improving the healing rate. This review describes the advances in sensors and biosensors for monitoring the concentration of C-reactive protein (CRP), temperature and pH in wounds. These three parameters can be used as qualitative biomarkers to assess the wound status and the effectiveness of therapy. CRP biosensors can be classified in: (a) field effect transistors, (b) optical immunosensors based on surface plasmon resonance, total internal reflection, fluorescence and chemiluminescence, (c) electrochemical sensors based on potentiometry, amperometry, and electrochemical impedance, and (d) piezoresistive sensors, such as quartz crystal microbalances and microcantilevers. The last section reports the most recent developments for wearable non-invasive temperature and pH sensors suitable for wound monitoring. PMID:29257113

Integrated multienzyme electrochemical biosensors for monitoring malolactic fermentation in wines.

PubMed

Gamella, M; Campuzano, S; Conzuelo, F; Curiel, J A; Muñoz, R; Reviejo, A J; Pingarrón, José M

2010-05-15

Integrated amperometric biosensors for the determination of L-malic and L-lactic acids were developed by coimmobilization of the enzymes L-malate dehydrogenase (MDH) and diaphorase (DP), or L-lactate oxidase (LOX) and horseradish peroxidase (HRP), respectively, together with the redox mediator tetrathiafulvalene (TTF), on a 3-mercaptopropionic acid (MPA) self-assembled monolayer (SAM)-modified gold electrode by using a dialysis membrane. The electrochemical oxidation of TTF at +100mV (vs. Ag/AgCl), and the reduction of TTF(+) at -50mV were used for the monitoring of the enzyme reactions involved in L-malic and L-lactic acid determinations, respectively. Experimental variables concerning the biosensors composition and the detection conditions were optimized for each biosensor. Good relative standard deviation values were obtained in both cases for the measurements carried out with the same biosensor, with no need of cleaning or pretreatment of the bioelectrodes surface, and with different biosensors constructed in the same manner. After 7 days of continuous use, the MDH/DP biosensor still exhibited 90% of the original sensitivity, while the LOX/HRP biosensor yielded a 91% of the original response after 5 days. Calibration graphs for L-malic and L-lactic were obtained with linear ranges of 5.2x10(-7) to 2.0x10(-5) and 4.2x10(-7) to 2.0x10(-5)M, respectively. The calculated detection limits were 5.2x10(-7) and 4.2x10(-7)M, respectively. The biosensors exhibited a high selectivity with no significant interferences. They were applied to monitor malolactic fermentation (MLF) induced by inoculation of Lactobacillus plantarum CECT 748(T) into a synthetic wine. Samples collected during MLF were assayed for L-malic and L-lactic acids, and the results obtained with the biosensors exhibited a very good correlation when plotted against those obtained by using commercial enzymatic kits.

Ring-Interferometric Sol-Gel Bio-Sensor

NASA Technical Reports Server (NTRS)

Bearman, Gregory (Inventor); Cohen, David (Inventor)

2006-01-01

A biosensor embodying the invention includes a sensing volume having an array of pores sized for immobilizing a first biological entity tending to bind to a second biological entity in such a manner as to change an index of refraction of the sensing volume. The biosensor further includes a ring interferometer, one volumetric section of the ring interferometer being the sensing volume, a laser for supplying light to the ring interferometer, and a photodetector for receiving light from the interferometer.

Selective detection of hypertoxic organophosphates pesticides via PDMS composite based acetylcholinesterase-inhibition biosensor.

PubMed

Zhao, Wei; Ge, Pei-Yu; Xu, Jing-Juan; Chen, Hong-Yuan

2009-09-01

We report on a pair of highly sensitive amperometric biosensors for organophosphate pesticides (OPs) based on assembling acetylcholinesterase (AChE) on poly(dimethylsiloxane) (PDMS)-poly(diallydimethylemmonium) (PDDA)/gold nanoparticles (AuNPs) composite film. Two AChE immobilization strategies are proposed based on the composite film with hydrophobic and hydrophilic surface tailored by oxygen plasma. The twin biosensors show interesting different electrochemical performances. The hydrophobic surface based PDMS-PDDAN AuNPs/choline oxidase (ChO)/AChE biosensor (biosensor-1) shows excellent stability and unique selectivity to hypertoxic organophosphate. At optimal conditions, this biosensor-1 could measure 5.0 x 10(-10) g/L paraoxon and 1.0 x 10(-9) g/L parathion. As for the hydrophilic surface based biosensor (biosensor-2), it shows no selectivity but can be commonly used for the detection of most OPs. Based on the structure of AChE, it is assumed that via the hydrophobic interaction between enzyme molecules and hydrophobic surface, the enzyme active sites surrounded by hydrophobic amino acids face toward the surface and get better protection from OPs. This assumption may explain the different performances of the twin biosensors and especially the unique selectivity of biosensor-1 to hypertoxic OPs. Real sample detection was performed and the omethoate residue on Cottomrose Hibiscus leaves was detected with biosensor-1.

Bacterial host and reporter gene optimization for genetically encoded whole cell biosensors.

PubMed

Brutesco, Catherine; Prévéral, Sandra; Escoffier, Camille; Descamps, Elodie C T; Prudent, Elsa; Cayron, Julien; Dumas, Louis; Ricquebourg, Manon; Adryanczyk-Perrier, Géraldine; de Groot, Arjan; Garcia, Daniel; Rodrigue, Agnès; Pignol, David; Ginet, Nicolas

2017-01-01

Whole-cell biosensors based on reporter genes allow detection of toxic metals in water with high selectivity and sensitivity under laboratory conditions; nevertheless, their transfer to a commercial inline water analyzer requires specific adaptation and optimization to field conditions as well as economical considerations. We focused here on both the influence of the bacterial host and the choice of the reporter gene by following the responses of global toxicity biosensors based on constitutive bacterial promoters as well as arsenite biosensors based on the arsenite-inducible P ars promoter. We observed important variations of the bioluminescence emission levels in five different Escherichia coli strains harboring two different lux-based biosensors, suggesting that the best host strain has to be empirically selected for each new biosensor under construction. We also investigated the bioluminescence reporter gene system transferred into Deinococcus deserti, an environmental, desiccation- and radiation-tolerant bacterium that would reduce the manufacturing costs of bacterial biosensors for commercial water analyzers and open the field of biodetection in radioactive environments. We thus successfully obtained a cell survival biosensor and a metal biosensor able to detect a concentration as low as 100 nM of arsenite in D. deserti. We demonstrated that the arsenite biosensor resisted desiccation and remained functional after 7 days stored in air-dried D. deserti cells. We also report here the use of a new near-infrared (NIR) fluorescent reporter candidate, a bacteriophytochrome from the magnetotactic bacterium Magnetospirillum magneticum AMB-1, which showed a NIR fluorescent signal that remained optimal despite increasing sample turbidity, while in similar conditions, a drastic loss of the lux-based biosensors signal was observed.

Biosensors engineered from conditionally stable ligand-binding domains

DOEpatents

Church, George M.; Feng, Justin; Mandell, Daniel J.; Baker, David; Fields, Stanley; Jester, Benjamin Ward; Tinberg, Christine Elaine

2017-09-19

Disclosed is a biosensor engineered to conditionally respond to the presence of specific small molecules, the biosensors including conditionally stable ligand-binding domains (LBDs) which respond to the presence of specific small molecules, wherein readout of binding is provided by reporter genes or transcription factors (TFs) fused to the LBDs.

Advances and challenges in biosensor-based diagnosis of infectious diseases

PubMed Central

Sin, Mandy LY; Mach, Kathleen E; Wong, Pak Kin; Liao, Joseph C

2014-01-01

Rapid diagnosis of infectious diseases and timely initiation of appropriate treatment are critical determinants that promote optimal clinical outcomes and general public health. Conventional in vitro diagnostics for infectious diseases are time-consuming and require centralized laboratories, experienced personnel and bulky equipment. Recent advances in biosensor technologies have potential to deliver point-of-care diagnostics that match or surpass conventional standards in regards to time, accuracy and cost. Broadly classified as either label-free or labeled, modern biosensors exploit micro- and nanofabrication technologies and diverse sensing strategies including optical, electrical and mechanical transducers. Despite clinical need, translation of biosensors from research laboratories to clinical applications has remained limited to a few notable examples, such as the glucose sensor. Challenges to be overcome include sample preparation, matrix effects and system integration. We review the advances of biosensors for infectious disease diagnostics and discuss the critical challenges that need to be overcome in order to implement integrated diagnostic biosensors in real world settings. PMID:24524681

Rapid sucrose monitoring in green coffee samples using multienzymatic biosensor.

PubMed

Stredansky, Miroslav; Redivo, Luca; Magdolen, Peter; Stredansky, Adam; Navarini, Luciano

2018-07-15

Amperometric biosensor utilizing FAD-dependent glucose dehydrogenase (FAD-GDH) for a specific sucrose monitoring in green coffee is described. FAD-GDH was co-immobilized with invertase and mutarotase on a thin-layer gold planar electrode using chitosan. The biosensor showed a wide linearity (from 10 to 1200 μM), low detection limit (8.4 μM), fast response time (50 s), and appeared to be O2 independent. In addition the biosensors exhibited a good operational (3 days) and storage (1 year) stability. Finally, the results achieved from the biosensor measurements of sucrose in 17 samples of green coffee (Coffea arabica, C. canephora and C. liberica) were compared with those obtained by the standard HPLC method. The good correlation among results of real samples, satisfactory analytical performance and simple use of the presented biosensor make it suitable for application in coffee industry. Copyright © 2018 Elsevier Ltd. All rights reserved.

Development of mercury (II) ion biosensors based on mercury-specific oligonucleotide probes.

PubMed

Li, Lanying; Wen, Yanli; Xu, Li; Xu, Qin; Song, Shiping; Zuo, Xiaolei; Yan, Juan; Zhang, Weijia; Liu, Gang

2016-01-15

Mercury (II) ion (Hg(2+)) contamination can be accumulated along the food chain and cause serious threat to the public health. Plenty of research effort thus has been devoted to the development of fast, sensitive and selective biosensors for monitoring Hg(2+). Thymine was demonstrated to specifically combine with Hg(2+) and form a thymine-Hg(2+)-thymine (T-Hg(2+)-T) structure, with binding constant even higher than T-A Watson-Crick pair in DNA duplex. Recently, various novel Hg(2+) biosensors have been developed based on T-rich Mercury-Specific Oligonucleotide (MSO) probes, and exhibited advanced selectivity and excellent sensitivity for Hg(2+) detection. In this review, we explained recent development of MSO-based Hg(2+) biosensors mainly in 3 groups: fluorescent biosensors, colorimetric biosensors and electrochemical biosensors. Copyright © 2015 Elsevier B.V. All rights reserved.

Fiber Optic Surface Plasmon Resonance-Based Biosensor Technique: Fabrication, Advancement, and Application.

PubMed

Liang, Gaoling; Luo, Zewei; Liu, Kunping; Wang, Yimin; Dai, Jianxiong; Duan, Yixiang

2016-05-03

Fiber optic-based biosensors with surface plasmon resonance (SPR) technology are advanced label-free optical biosensing methods. They have brought tremendous progress in the sensing of various chemical and biological species. This review summarizes four sensing configurations (prism, grating, waveguide, and fiber optic) with two ways, attenuated total reflection (ATR) and diffraction, to excite the surface plasmons. Meanwhile, the designs of different probes (U-bent, tapered, and other probes) are also described. Finally, four major types of biosensors, immunosensor, DNA biosensor, enzyme biosensor, and living cell biosensor, are discussed in detail for their sensing principles and applications. Future prospects of fiber optic-based SPR sensor technology are discussed.

Recent research trends of radio-frequency biosensors for biomolecular detection.

PubMed

Lee, Hee-Jo; Yook, Jong-Gwan

2014-11-15

This article reviews radio-frequency (RF) biosensors based on passive and/or active devices and circuits. In particular, we focus on RF biosensors designed for detection of various biomolecules such as biotin-streptavidin, DNA hybridization, IgG, and glucose. The performance of these biosensors has been enhanced by the introduction of various sensing schemes with diverse nanomaterials (e.g., carbon nanotubes, graphene oxide, magnetic and gold nanoparticles, etc.). In addition, the RF biosensing platforms that can be associated with an RF active system are discussed. Finally, the challenges of RF biosensors are presented and suggestions are made for their future direction and prospects. Copyright © 2014 Elsevier B.V. All rights reserved.

Integrated multienzyme electrochemical biosensors for the determination of glycerol in wines.

PubMed

Gamella, M; Campuzano, S; Reviejo, A J; Pingarrón, J M

2008-02-25

The construction and performance of integrated amperometric biosensors for the determination of glycerol are reported. Two different biosensor configurations have been evaluated: one based on the glycerol dehydrogenase/diaphorase (GDH/DP) bienzyme system, and another using glycerol kinase/glycerol-3-phosphate oxidase/peroxidase (GK/GPOx/HRP). Both enzyme systems were immobilized together with the mediator tetrathiafulvalene (TTF) on a 3-mercaptopropionic acid (MPA) self-assembled monolayer (SAM)-modified gold electrode by using a dialysis membrane. The electrochemical oxidation of TTF at +150mV (vs. Ag/AgCl), and the reduction of TTF(+) at 0mV were used for the monitoring of the enzyme reactions for the bienzyme and trienzyme configurations, respectively. Experimental variables concerning both the biosensors composition and the working conditions were optimized for each configuration. A good repeatability of the measurements with no need of cleaning or pretreatment of the biosensors was obtained in both cases. After 51 days of use, the GDH/DP biosensor still exhibited 87% of the original sensitivity, while the GK/GPOx/HRP biosensor yielded a 46% of the original response after 8 days. Calibration graphs for glycerol with linear ranges of 1.0x10(-6) to 2.0x10(-5) or 1.0x10(-6) to 1.0x10(-5)M glycerol and sensitivities of 1214+/-21 or 1460+/-34microAM(-1) were obtained with GDH/DP and GK/GPOx/HRP biosensors, respectively. The calculated detection limits were 4.0x10(-7) and 3.1x10(-7)M, respectively. The biosensors exhibited a great sensitivity with no significant interferences in the analysis of wines. The biosensors were applied to the determination of glycerol in 12 different wines and the results advantageously compared with those provided by a commercial enzyme kit.

Advances in the manufacturing, types, and applications of biosensors

NASA Astrophysics Data System (ADS)

Ravindra, Nuggehalli M.; Prodan, Camelia; Fnu, Shanmugamurthy; Padronl, Ivan; Sikha, Sushil K.

2007-12-01

In recent years, there have been significant technological advancements in the manufacturing, types, and applications of biosensors. Applications include clinical and non-clinical diagnostics for home, bio-defense, bio-remediation, environment, agriculture, and the food industry. Biosensors have progressed beyond the detection of biological threats such as anthrax and are finding use in a number of non-biological applications. Emerging biosensor technologies such as lab-on-a-chip have revolutionized the integration approaches for a very flexible, innovative, and user-friendly platform. An overview of the fundamentals, types, applications, and manufacturers, as well as the market trends of biosensors is presented here. Two case studies are discussed: one focused on a characterization technique—patch clamping and dielectric spectroscopy as a biological sensor—and the other about lithium phthalocyanine, a material that is being developed for in-vivo oxymetry.

Environmental Stability of Plasmonic Biosensors Based on Natural versus Artificial Antibody.

PubMed

Luan, Jingyi; Xu, Ting; Cashin, John; Morrissey, Jeremiah J; Kharasch, Evan D; Singamaneni, Srikanth

2018-06-13

Plasmonic biosensors based on the refractive index sensitivity of localized surface plasmon resonance (LSPR) are considered to be highly promising for on-chip and point-of-care biodiagnostics. However, most of the current plasmonic biosensors employ natural antibodies as biorecognition elements, which can easily lose their biorecognition ability upon exposure to environmental stressors (e.g., temperature and humidity). Plasmonic biosensors relying on molecular imprints as recognition elements (artificial antibodies) are hypothesized to be an attractive alternative for applications in resource-limited settings due to their excellent thermal, chemical, and environmental stability. In this work, we provide a comprehensive comparison of the stability of plasmonic biosensors based on natural and artificial antibodies. Although the natural antibody-based plasmonic biosensors exhibit superior sensitivity, their stability (temporal, thermal, and chemical) was found to be vastly inferior to those based on artificial antibodies. Our results convincingly demonstrate that these novel classes of artificial antibody-based plasmonic biosensors are highly attractive for point-of-care and resource-limited conditions where tight control over transport, storage, and handling conditions is not possible.

Nanomaterials towards fabrication of cholesterol biosensors: Key roles and design approaches.

PubMed

Saxena, Urmila; Das, Asim Bikas

2016-01-15

Importance of cholesterol biosensors is already recognized in the clinical diagnosis of cardiac and brain vascular diseases as discernible from the enormous amount of research in this field. Nevertheless, the practical application of a majority of the fabricated cholesterol biosensors is ordinarily limited by their inadequate performance in terms of one or more analytical parameters including stability, sensitivity and detection limit. Nanoscale materials offer distinctive size tunable electronic, catalytic and optical properties which opened new opportunities for designing highly efficient biosensor devices. Incorporation of nanomaterials in biosensing devices has found to improve the electroactive surface, electronic conductivity and biocompatibility of the electrode surfaces which then improves the analytical performance of the biosensors. Here we have reviewed recent advances in nanomaterial-based cholesterol biosensors. Foremost, the diverse roles of nanomaterials in these sensor systems have been discussed. Later, we have exhaustively explored the strategies used for engineering cholesterol biosensors with nanotubes, nanoparticles and nanocomposites. Finally, this review concludes with future outlook signifying some challenges of these nanoengineered cholesterol sensors. Copyright © 2015 Elsevier B.V. All rights reserved.

A Highly Responsive Silicon Nanowire/Amplifier MOSFET Hybrid Biosensor

DTIC Science & Technology

2015-07-21

biosensor. The insets show a magnified view of the SiNW channel region (W = 55 nm). ( c ) Photograph of the biosensor chip fabricated via a top-down method...of the SiNW FET is 147 mV/decade. (b) VT and ( c ) ISINW at different pH levels; these values were extracted from Fig. 2a. VT was extracted using the...function of pH level in the hybrid biosensor. The extracted current change is 5.5 × 105 (=5.74 decade per pH). ( c ) Transient response of IMOSFET while

Oxygen sensing glucose biosensors based on alginate nano-micro systems

NASA Astrophysics Data System (ADS)

Chaudhari, Rashmi; Joshi, Abhijeet; Srivastava, Rohit

2014-04-01

Clinically glucose monitoring in diabetes management is done by point-measurement. However, an accurate, continuous glucose monitoring, and minimally invasive method is desirable. The research aims at developing fluorescence-mediated glucose detecting biosensors based on near-infrared radiation (NIR) oxygen sensitive dyes. Biosensors based on Glucose oxidase (GOx)-Rudpp loaded alginate microspheres (GRAM) and GOx-Platinum-octaethylporphyrin (PtOEP)-PLAalginate microsphere system (GPAM) were developed using air-driven atomization and characterized using optical microscopy, CLSM, fluorescence spectro-photometry etc. Biosensing studies were performed by exposing standard solutions of glucose. Uniform sized GRAM and GPAM with size 50+/-10μm were formed using atomization. CLSM imaging of biosensors suggests that Rudpp and PtOEP nanoparticles are uniformly distributed in alginate microspheres. The GRAM and GPAM showed a good regression constant of 0.974 and of 0.9648 over a range of 0-10 mM of glucose with a high sensitivity of 3.349%/mM (625 nm) and 2.38%/mM (645 nm) at 10 mM of glucose for GRAM and GPAM biosensor. GRAM and GPAM biosensors show great potential in development of an accurate and minimally invasive glucose biosensor. NIR dye based assays can aid sensitive, minimally-invasive and interference-free detection of glucose in diabetic patients.

Rapid solid-phase immunoassay for 6-keto prostaglandin F1 alpha on microplates

DOE Office of Scientific and Technical Information (OSTI.GOV)

Schramm, W.; Smith, R.H.; Jackson, T.M.

1990-03-01

We describe, for the measurement of 6-keto prostaglandin F1 alpha in biological media, a solid-phase immunoassay with immobilized antibodies that requires a total processing time of less than 2 h with hands-on time less than 30 min for 40 samples. The method combines the convenience of the microplate format with the sensitivity of radiolabeled prostaglandin derivatives as tracers in a competitive immunoassay. The intra- and interassay variations at 50% displacement of the radiolabeled prostaglandin derivative as tracer were 9.0% and 11.8%, respectively. At 50% displacement of the radiolabeled tracer, the sensitivity is about 20 pg per well. Optimal incubation timemore » is between 60 and 90 min. Nonspecific binding was less than 1% if about 8 pg of tracer (approximately 25,000 counts/min per well) was used. Inhibition curves of samples in different dilutions were parallel to standard curves. The variation of bound radiolabeled prostaglandin derivative within the wells of one microplate (n = 96) was less than 3%. Human plasma samples and medium from tissue culture assayed for 6-keto prostaglandin F1 alpha correlated well with results obtained with a solid-phase assay based on use of magnetic particles (r = 0.99, n = 24) for culture-medium samples; r = 0.99; n = 26 for plasma samples.« less

Brunn: an open source laboratory information system for microplates with a graphical plate layout design process.

PubMed

Alvarsson, Jonathan; Andersson, Claes; Spjuth, Ola; Larsson, Rolf; Wikberg, Jarl E S

2011-05-20

Compound profiling and drug screening generates large amounts of data and is generally based on microplate assays. Current information systems used for handling this are mainly commercial, closed source, expensive, and heavyweight and there is a need for a flexible lightweight open system for handling plate design, and validation and preparation of data. A Bioclipse plugin consisting of a client part and a relational database was constructed. A multiple-step plate layout point-and-click interface was implemented inside Bioclipse. The system contains a data validation step, where outliers can be removed, and finally a plate report with all relevant calculated data, including dose-response curves. Brunn is capable of handling the data from microplate assays. It can create dose-response curves and calculate IC50 values. Using a system of this sort facilitates work in the laboratory. Being able to reuse already constructed plates and plate layouts by starting out from an earlier step in the plate layout design process saves time and cuts down on error sources.

A novel immobilization multienzyme glucose fluorescence capillary biosensor.

PubMed

Li, Yong-Sheng; Du, Yun-Dong; Chen, Ting-Mei; Gao, Xiu-Feng

2010-02-15

Based on the immobilization enzyme technology and the fluorescence capillary analysis method, the authors have developed a highly sensitive fluorescence reaction system and a novel immobilization multienzyme glucose fluorescence capillary biosensor for determining glucose contents. Reaction principle of the system is that under the catalysis of glucose oxidase (GOD) and horseradish peroxidase (HRP) immobilized on inner surface of a medical capillary, beta-D-glucose reacts with dissolved oxygen to form gluconic acid-delta-lactone and hydrogen peroxide, and then the latter reacts with l-tyrosine to produce a tyrosine dimer, which has maximal excitation and emission wavelengths at 320 nm and 410 nm, respectively. Fluorescence of the dimer is proportional to the concentration of the beta-D-glucose. Optimization conditions suitable for the reaction system and the biosensor were as follows. Concentration of the L-tyrosine used as fluorescence reagent was 0.15 mol L(-1), the active concentrations of the GOD and the HRP for the immobilization were 15 kU L(-1) and 8 kU L(-1), respectively. Consumptions of the sample and reagents in one determination were 5.0 microL and 15 microL, respectively. Quantitative range of the biosensor for the glucose was in the range 1-10 micromol L(-1), its relative standard deviation was less than 4.9%, and its detection limit was 0.62 micromol L(-1). The biosensor's recovery was in the range 96-105%. Results of some serum determined with the biosensor and with a commercial glucose-kit were well coincident to each other. Accordingly, the biosensor can be applied to the determination of serum glucose contents in the diagnosis of diabetes. Copyright 2009 Elsevier B.V. All rights reserved.

Electrochemical Aptamer Scaffold Biosensors for Detection of Botulism and Ricin Proteins.

PubMed

Daniel, Jessica; Fetter, Lisa; Jett, Susan; Rowland, Teisha J; Bonham, Andrew J

2017-01-01

Electrochemical DNA (E-DNA) biosensors enable the detection and quantification of a variety of molecular targets, including oligonucleotides, small molecules, heavy metals, antibodies, and proteins. Here we describe the design, electrode preparation and sensor attachment, and voltammetry conditions needed to generate and perform measurements using E-DNA biosensors against two protein targets, the biological toxins ricin and botulinum neurotoxin. This method can be applied to generate E-DNA biosensors for the detection of many other protein targets, with potential advantages over other systems including sensitive detection limits typically in the nanomolar range, real-time monitoring, and reusable biosensors.

A Colorimetric Microplate Assay for DNA-Binding Activity of His-Tagged MutS Protein.

PubMed

Banasik, Michał; Sachadyn, Paweł

2016-09-01

A simple microplate method was designed for rapid testing DNA-binding activity of proteins. The principle of the assay involves binding of tested DNA by his-tagged protein immobilized on a nickel-coated ELISA plate, following colorimetric detection of biotinylated DNA with avidin conjugated to horseradish peroxidase. The method was used to compare DNA mismatch binding activities of MutS proteins from three bacterial species. The assay required relatively low amounts of tested protein (approximately 0.5-10 pmol) and DNA (0.1-10 pmol) and a relatively short time of analysis (up to 60 min). The method is very simple to apply and convenient to test different buffer conditions of DNA-protein binding. Sensitive colorimetric detection enables naked eye observations and quantitation with an ELISA reader. The performance of the assay, which we believe is a distinguishing trait of the method, is based on two strong and specific molecular interactions: binding of a his-tagged protein to a nickel-coated microplate and binding of biotinylated DNA to avidin. In the reported experiments, the solution was used to optimize the conditions for DNA mismatch binding by MutS protein; however, the approach could be implemented to test nucleic acids interactions with any protein of interest.

A Sensitive DNA Capacitive Biosensor Using Interdigitated Electrodes

PubMed Central

Wang, Lei; Veselinovic, Milena; Yang, Lang; Geiss, Brian J.; Dandy, David S.; Chen, Tom

2017-01-01

This paper presents a label-free affinity-based capacitive biosensor using interdigitated electrodes. Using an optimized process of DNA probe preparation to minimize the effect of contaminants in commercial thiolated DNA probe, the electrode surface was functionalized with the 24-nucleotide DNA probes based on the West Nile virus sequence (Kunjin strain). The biosensor has the ability to detect complementary DNA fragments with a detection limit down to 20 DNA target molecules (1.5 aM range), making it suitable for a practical point-of-care (POC) platform for low target count clinical applications without the need for amplification. The reproducibility of the biosensor detection was improved with efficient covalent immobilization of purified single-stranded DNA probe oligomers on cleaned gold microelectrodes. In addition to the low detection limit, the biosensor showed a dynamic range of detection from 1 μL−1 to 105 μL−1 target molecules (20 to 2 million targets), making it suitable for sample analysis in a typical clinical application environment. The binding results presented in this paper were validated using fluorescent oligomers. PMID:27619528

The blocking reagent optimization for the magnetoelastic biosensor

NASA Astrophysics Data System (ADS)

Hu, Jiajia; Chai, Yating; Horikawa, Shin; Wikle, Howard C.; Wang, Feng'en; Du, Songtao; Chin, Bryan A.; Hu, Jing

2015-06-01

The wireless phage-based magnetoelastic (ME) biosensor has proven to be promising for real-time detection of pathogenic bacteria on fresh produces. The ME biosensor consists of a freestanding ME resonator as the signal transducer and filamentous phage as the biomolecular-recognition element, which can specifically bind to a pathogen of interest. Due to the Joule magnetostriction effect, the biosensors can be placed into mechanical resonance when subjected to a time-varying magnetic field alternating at the sensor's resonant frequency. Upon the attachment of the target pathogen, the mass of the biosensor increases, thereby decreasing its resonant frequency. This paper presents an investigation of blocking reagents immobilization for detecting Salmonella Typhimurium on fresh food surfaces. Three different blocking reagents (BSA, SuperBlock blocking buffer, and blocker BLOTTO) were used and compared. The optical microscope was used for bacterial cells binding observation. Student t-test was used to statistically analysis the experiment results. The results shows that SuperBlock blocking buffer and blocker BLOTTO have much better blocking performance than usually used BSA.

Prediction of the limit of detection of an optical resonant reflection biosensor.

PubMed

Hong, Jongcheol; Kim, Kyung-Hyun; Shin, Jae-Heon; Huh, Chul; Sung, Gun Yong

2007-07-09

A prediction of the limit of detection of an optical resonant reflection biosensor is presented. An optical resonant reflection biosensor using a guided-mode resonance filter is one of the most promising label-free optical immunosensors due to a sharp reflectance peak and a high sensitivity to the changes of optical path length. We have simulated this type of biosensor using rigorous coupled wave theory to calculate the limit of detection of the thickness of the target protein layer. Theoretically, our biosensor has an estimated ability to detect thickness change approximately the size of typical antigen proteins. We have also investigated the effects of the absorption and divergence of the incident light on the detection ability of the biosensor.

Inkjet printing for biosensor fabrication: combining chemistry and technology for advanced manufacturing.

PubMed

Li, Jia; Rossignol, Fabrice; Macdonald, Joanne

2015-06-21

Inkjet printing is emerging at the forefront of biosensor fabrication technologies. Parallel advances in both ink chemistry and printers have led to a biosensor manufacturing approach that is simple, rapid, flexible, high resolution, low cost, efficient for mass production, and extends the capabilities of devices beyond other manufacturing technologies. Here we review for the first time the factors behind successful inkjet biosensor fabrication, including printers, inks, patterning methods, and matrix types. We discuss technical considerations that are important when moving beyond theoretical knowledge to practical implementation. We also highlight significant advances in biosensor functionality that have been realised through inkjet printing. Finally, we consider future possibilities for biosensors enabled by this novel combination of chemistry and technology.

Pulsed excitation system to measure the resonant frequency of magnetoelastic biosensors

NASA Astrophysics Data System (ADS)

Xie, Hong; Chai, Yating; Horikawa, Shin; Wikle, Howard C.; Chin, Bryan A.

2014-05-01

An electrical circuit was designed and tested to measure the resonant frequency of micron-scale magnetoelastic (ME) biosensors using a pulsed wave excitation technique. In this circuit, a square pulse current is applied to an excitation coil to excite the vibration of ME biosensors and a pick-up coil is used to sense the ME biosensor's mechanical vibration and convert it to an electrical output signal. The output signal is filtered and amplified by a custom designed circuit to allow the measurement of the resonant frequency of the ME biosensor from which the detection of specific pathogens can be made. As a proof-in-concept experiment, JRB7 phage-coated ME biosensors were used to detect different concentrations of Bacillus anthracis Sterne strain spores. A statistically significant difference was observed for concentrations of 5 × 102 spore/ml and above.

[Establishment of an in vitro screening model for steroid 5 alpha-reductase inhibitors with the microplate reader].

PubMed

Wu, Jian-Hui; Sun, Zu-Yue

2013-06-01

To establish an in vitro screening model for steroid 5 alpha-reductase inhibitors using the microplate reader. Steroid 5 alpha-reductase was obtained from the liver of female rats, an in vitro screening model for steroid 5 alpha-reductase inhibitors established using the 96-well plate and microplate reader after determination of the enzymatic activity, and the reliability of the model verified with the known 5 alpha-reductase inhibitors epristeride and finasteride. Added to the 96-well plate were the final concentrations of testosterone (0-40 micromol/L), NADPH (22 micromol/L), epristeride (0-60 nmol/L) or finasteride (0-60 nmol/ L) and steroid 5 alpha-reductase (20 microl), the total volume of each well adjusted to 200 microl with Tris-Hcl buffer. The 96-well plate was placed in the microplate reader, mixed and incubated at 37 degrees C, followed by detection of the A340nm value at 0 and 10 min and analysis of the data. The Km value of steroid 5 alpha-reductase was 3.794 micromol/L, with a Vmax of 0.271 micromol/(L. min). The Ki of epristeride was 148.2 nmol/L, with an IC50 of 31.5 nmol/L, and the enzymatic reaction kinetic curve suggested that epristeride was an uncompetitive enzyme inhibitor. The Ki of finasteride was 158. 8 nmol/L, with an IC50 of 13.6 nmol/L. The enzymatic reaction kinetic curve showed that both epristeride and finasteride were competitive enzyme inhibitors, similar to those reported in the published literature. A screening model was successfully established, which could rapidly and effectively screen steroid 5 alpha-reductase inhibitors in vitro.

Microbially derived biosensors for diagnosis, monitoring and epidemiology.

PubMed

Chang, Hung-Ju; Voyvodic, Peter L; Zúñiga, Ana; Bonnet, Jérôme

2017-09-01

Living cells have evolved to detect and process various signals and can self-replicate, presenting an attractive platform for engineering scalable and affordable biosensing devices. Microbes are perfect candidates: they are inexpensive and easy to manipulate and store. Recent advances in synthetic biology promise to streamline the engineering of microbial biosensors with unprecedented capabilities. Here we review the applications of microbially-derived biosensors with a focus on environmental monitoring and healthcare applications. We also identify critical challenges that need to be addressed in order to translate the potential of synthetic microbial biosensors into large-scale, real-world applications. © 2017 The Authors. Microbial Biotechnology published by John Wiley & Sons Ltd and Society for Applied Microbiology.

Potentiometric urea biosensor based on an immobilised fullerene-urease bio-conjugate.

PubMed

Saeedfar, Kasra; Heng, Lee Yook; Ling, Tan Ling; Rezayi, Majid

2013-12-06

A novel method for the rapid modification of fullerene for subsequent enzyme attachment to create a potentiometric biosensor is presented. Urease was immobilized onto the modified fullerene nanomaterial. The modified fullerene-immobilized urease (C60-urease) bioconjugate has been confirmed to catalyze the hydrolysis of urea in solution. The biomaterial was then deposited on a screen-printed electrode containing a non-plasticized poly(n-butyl acrylate) (PnBA) membrane entrapped with a hydrogen ionophore. This pH-selective membrane is intended to function as a potentiometric urea biosensor with the deposition of C60-urease on the PnBA membrane. Various parameters for fullerene modification and urease immobilization were investigated. The optimal pH and concentration of the phosphate buffer for the urea biosensor were 7.0 and 0.5 mM, respectively. The linear response range of the biosensor was from 2.31 × 10-3 M to 8.28 × 10-5 M. The biosensor's sensitivity was 59.67 ± 0.91 mV/decade, which is close to the theoretical value. Common cations such as Na+, K+, Ca2+, Mg2+ and NH4+ showed no obvious interference with the urea biosensor's response. The use of a fullerene-urease bio-conjugate and an acrylic membrane with good adhesion prevented the leaching of urease enzyme and thus increased the stability of the urea biosensor for up to 140 days.

Did the Kyrenia Range of northern Cyprus rotate with the Troodos-Hatay microplate during the tectonic evolution of the eastern Mediterranean?

NASA Astrophysics Data System (ADS)

Morris, Antony; Robertson, Alastair H. F.; Anderson, Mark W.; Hodgson, Emma

2016-01-01

Previous palaeomagnetic studies have allowed the recognition of a distinctive area of Neotethyan oceanic rocks, including the Troodos ophiolite in Cyprus and the Hatay ophiolite to the east in southern Turkey, that underwent 90° of anticlockwise rotation between Late Cretaceous (Campanian) and Early Eocene time. The southern and western boundaries of this rotated Troodos-Hatay microplate have been inferred to lie within, or adjacent to, zones of deformed oceanic and continental margin rocks that are now exposed in southern and western Cyprus; however, the northern boundary of the microplate remains undefined. Relevant to this problem, palaeomagnetic data are presented here from basaltic lavas exposed along the Kyrenia Range, mostly from Late Cretaceous (Maastrichtian) sites and one Eocene site. A positive inclination-only fold test demonstrates that remanences are pre-deformational in age, and positive conglomerate tests show that magnetic remanences were acquired before Late Eocene-Early Oligocene time, together suggesting that primary magnetizations are preserved. Data from the eastern Kyrenia Range and the Karpas Peninsula (the easternmost extension of the Kyrenia Range) document significant relative tectonic rotation between these localities, with no rotation in the eastern range versus 30° of anticlockwise rotation of the Karpas Peninsula. Unfortunately, palaeomagnetic sites from the western Kyrenia Range did not yield tectonically interpretable magnetization directions, probably due to complex poly-phase thrusting and folding, and the central range also yielded no interpretable data. However, the available palaeomagnetic data are sufficient to demonstrate that the Kyrenia terrane underwent a separate rotation history to the Troodos-Hatay microplate and also implies that the northern boundary of the Troodos-Hatay microplate was located between the Troodos ophiolite and the Kyrenia Range. The former microplate margin has since been overridden and concealed by

Measurement of the distribution of non-structural carbohydrate composition in onion populations by a high-throughput microplate enzymatic assay.

PubMed

Revanna, Roopashree; Turnbull, Matthew H; Shaw, Martin L; Wright, Kathryn M; Butler, Ruth C; Jameson, Paula E; McCallum, John A

2013-08-15

Non-structural carbohydrate (NSC; glucose, fructose, sucrose and fructan) composition of onions (Allium cepa L.) varies widely and is a key determinant of market usage. To analyse the physiology and genetics of onion carbohydrate metabolism and to enable selective breeding, an inexpensive, reliable and practicable sugar assay is required to phenotype large numbers of samples. A rapid, reliable and cost-effective microplate-based assay was developed for NSC analysis in onions and used to characterise variation in tissue hexose, sucrose and fructan content in open-pollinated breeding populations and in mapping populations developed from a wide onion cross. Sucrose measured in microplates employing maltase as a hydrolytic enzyme was in agreement with HPLC-PAD results. The method revealed significant variation in bulb fructan content within open-pollinated 'Pukekohe Longkeeper' breeding populations over a threefold range. Very wide segregation from 80 to 600 g kg(-1) in fructan content was observed in bulbs of F2 genetic mapping populations from the wide onion cross 'Nasik Red × CUDH2150'. The microplate enzymatic assay is a reliable and practicable method for onion sugar analysis for genetics, breeding and food technology. Open-pollinated onion populations may harbour extensive within-population variability in carbohydrate content, which may be quantified and exploited using this method. The phenotypic data obtained from genetic mapping populations show that the method is well suited to detailed genetic and physiological analysis. © 2013 Society of Chemical Industry.

Label-free, real-time interaction and adsorption analysis 2: quartz crystal microbalance.

PubMed

Fee, Conan J

2013-01-01

In this chapter, a second biosensor technique is described: the quartz crystal microbalance (QCM). The quartz crystal microbalance is a physical technique that detects changes in the resonance frequency of an electrically driven quartz crystal with changes in mass. Unlike surface plasmon resonance (SPR), QCM is affected by both the water that may be associated with the adsorbed layer and by conformational changes in the adsorbed species, while SPR is insensitive to both effects. Thus QCM can both corroborate the findings of an SPR experiment and provide some complementary information. Also, the QCM surface is highly versatile and can range from plain quartz, through gold and other metal surfaces (e.g., titanium or stainless steel) to polymeric materials. Thus, the QCM technique has wide utility in tracking interactions with a variety of materials.

Development of conductometric biosensor array for simultaneous determination of maltose, lactose, sucrose and glucose.

PubMed

Soldatkin, O O; Peshkova, V M; Saiapina, O Y; Kucherenko, I S; Dudchenko, O Y; Melnyk, V G; Vasylenko, O D; Semenycheva, L M; Soldatkin, A P; Dzyadevych, S V

2013-10-15

The aim of this work was to develop an array of biosensors for simultaneous determination of four carbohydrates in solution. Several enzyme systems selective to lactose, maltose, sucrose and glucose were immobilised on the surface of four conductometric transducers and served as bio-recognition elements of the biosensor array. Direct enzyme analysis carried out by the developed biosensors was highly sensitive to the corresponding substrates. The analysis lasted 2 min. The dynamic range of substrate determination extended from 0.001 mM to 1.0-3.0mM, and strongly depended on the enzyme system used. An effect of the solution pH, ionic strength and buffer capacity on the biosensors responses was investigated; the conditions of simultaneous operation of all biosensors were optimised. The data on cross-impact of the substrates of all biosensors were obtained; the biosensor selectivity towards possible interfering carbohydrates was tested. The developed biosensor array showed good signal reproducibility and storage stability. The biosensor array is suited for simultaneous, quick, simple, and selective determination of maltose, lactose, sucrose and glucose. © 2013 Elsevier B.V. All rights reserved.

Detection of rabbit IgG by using functional magnetic particles and an enzyme-conjugated antibody with a homemade magnetic microplate.

PubMed

Tsai, Hweiyan; Lu, Yi-Hsuan; Liao, Huan-Xuan; Wu, Shih-Wei; Yu, Feng-Yih; Fuh, Chwan Bor

2015-01-01

The enzyme-linked immunosorbent assay (ELISA) has been used for diagnosing medical and plant pathologies. In addition, it is used for quality-control evaluations in various industries. The ELISA is the simplest method for obtaining excellent results; however, it is time consuming because the immunoreagents interact only on the contact surfaces. Antibody-labeled magnetic particles can be dispersed in a solution to yield a pseudohomogeneous reaction with antigens which improved the efficiency of immunoreaction, and can be easily separated from the unreactive substances by applying a magnetic force. We used a homemade magnetic microplate, functional magnetic particles (MPs) and enzyme-labeled secondary antibody to perform the sandwich ELISA successfully. Using antibody-labeled MPs enabled reducing the analysis time to one-third of that required in using a conventional ELISA. The secondary antibody conjugated with horseradish peroxidase (HRP) was affinity-bound to the analyte (IgG in this study). The calibration curve was established according to the measured absorbance of the 3, 3', 5, 5'-tetramethybezidine-HRP reaction products versus the concentrations of standard IgG. The linear range of IgG detection was 114 ng/mL-3.5 ng/mL. The limit of detection (LOD) of IgG was 3.4 ng/mL. The recovery and coefficient of variation were 100% (±7%) and 116% (±4%) for the spiked concentrations of 56.8 ng/mL and 14.2 ng/mL, respectively. Pseudohomogeneous reactions can be performed using functional MPs and a magnetic microplate. Using antibody-labeled MPs, the analysis time can be reduced to one-third of that required in using a conventional ELISA. The substrate-enzyme reaction products can be easily transferred to another microplate, and their absorbance can be measured without interference by light scattering caused by magnetic microbeads. This method demonstrates great potential for detecting other biomarkers and in biochemical applications. Graphical AbstractA magnetic

PEGylation of a Maltose Biosensor Promotes Enhanced Signal Response

DOE Office of Scientific and Technical Information (OSTI.GOV)

Dattelbaum, Andrew; Baker, Gary A; Fox, John M

2009-01-01

A robust method to immobilize a maltose biosensor is described using an engineered maltose periplasmic binding protein (PBP) covalently coupled to NBDamide, an environmentally sensitive fluorophore. A mesoporous silica sol-gel derived from diglycerylsilane (DGS) was constructed to embed the maltose biosensor, and the ligand reporting fluorescence properties were meas red. When sequestered in the DGS-derived silica matrix, the biosensor retained maltose-dependent fluorescence sensing capability with micromolar affinity, which is consistent with the protein free in solution. The MBP-NBD conjugate was further modified by covalent conjugation with poly(ethylene glycol)-5000 (PEG) to promote the retention of water molecules around the protein andmore » to reduce possible steric effects between the silica matrix and protein. Bioconjugation with PEG molecules does not significantly affect the signaling response of the protein in solution. When immobilized in the DGS polymer, a consistent increase in fluorescence intensity was observed as compared to the protein not functionalized with PEG. To our knowledge, this report presents the first successful method to embed a PBP biosensor in a polymerized matrix and retain signaling response using an environmentally sensitive probe. The immobilization method presented here should be easily adaptable to all conformation-dependent biosensors.« less

Biosensors and bioelectronics on smartphone for portable biochemical detection.

PubMed

Zhang, Diming; Liu, Qingjun

2016-01-15

Smartphone has been widely integrated with sensors, such as test strips, sensor chips, and hand-held detectors, for biochemical detections due to its portability and ubiquitous availability. Utilizing built-in function modules, smartphone is often employed as controller, analyzer, and displayer for rapid, real-time, and point-of-care monitoring, which can significantly simplify design and reduce cost of the detecting systems. This paper presents a review of biosensors and bioelectronics on smartphone for portable biochemical detections. The biosensors and bioelectronics based on smartphone can mainly be classified into biosensors using optics, surface plasmon resonance, electrochemistry, and near-field communication. The developments of these biosensors and bioelectronics on smartphone are reviewed along with typical biochemical detecting cases. Sensor strategies, detector attachments, and coupling methods are highlighted to show designs of the compact, lightweight, and low-cost sensor systems. The performances and advantages of these designs are introduced with their applications in healthcare diagnosis, environment monitoring, and food evaluation. With advances in micro-manufacture, sensor technology, and miniaturized electronics, biosensor and bioelectronic devices on smartphone can be used to perform biochemical detections as common and convenient as electronic tag readout in foreseeable future. Copyright © 2015 Elsevier B.V. All rights reserved.

Microbial Fuels Cell-Based Biosensor for Toxicity Detection: A Review

PubMed Central

Zhou, Tuoyu; Han, Huawen; Liu, Pu; Xiong, Jian; Tian, Fake; Li, Xiangkai

2017-01-01

With the unprecedented deterioration of environmental quality, rapid recognition of toxic compounds is paramount for performing in situ real-time monitoring. Although several analytical techniques based on electrochemistry or biosensors have been developed for the detection of toxic compounds, most of them are time-consuming, inaccurate, or cumbersome for practical applications. More recently, microbial fuel cell (MFC)-based biosensors have drawn increasing interest due to their sustainability and cost-effectiveness, with applications ranging from the monitoring of anaerobic digestion process parameters (VFA) to water quality detection (e.g., COD, BOD). When a MFC runs under correct conditions, the voltage generated is correlated with the amount of a given substrate. Based on this linear relationship, several studies have demonstrated that MFC-based biosensors could detect heavy metals such as copper, chromium, or zinc, as well as organic compounds, including p-nitrophenol (PNP), formaldehyde and levofloxacin. Both bacterial consortia and single strains can be used to develop MFC-based biosensors. Biosensors with single strains show several advantages over systems integrating bacterial consortia, such as selectivity and stability. One of the limitations of such sensors is that the detection range usually exceeds the actual pollution level. Therefore, improving their sensitivity is the most important for widespread application. Nonetheless, MFC-based biosensors represent a promising approach towards single pollutant detection. PMID:28956857

Microbial Fuels Cell-Based Biosensor for Toxicity Detection: A Review.

PubMed

Zhou, Tuoyu; Han, Huawen; Liu, Pu; Xiong, Jian; Tian, Fake; Li, Xiangkai

2017-09-28

With the unprecedented deterioration of environmental quality, rapid recognition of toxic compounds is paramount for performing in situ real-time monitoring. Although several analytical techniques based on electrochemistry or biosensors have been developed for the detection of toxic compounds, most of them are time-consuming, inaccurate, or cumbersome for practical applications. More recently, microbial fuel cell (MFC)-based biosensors have drawn increasing interest due to their sustainability and cost-effectiveness, with applications ranging from the monitoring of anaerobic digestion process parameters (VFA) to water quality detection (e.g., COD, BOD). When a MFC runs under correct conditions, the voltage generated is correlated with the amount of a given substrate. Based on this linear relationship, several studies have demonstrated that MFC-based biosensors could detect heavy metals such as copper, chromium, or zinc, as well as organic compounds, including p -nitrophenol (PNP), formaldehyde and levofloxacin. Both bacterial consortia and single strains can be used to develop MFC-based biosensors. Biosensors with single strains show several advantages over systems integrating bacterial consortia, such as selectivity and stability. One of the limitations of such sensors is that the detection range usually exceeds the actual pollution level. Therefore, improving their sensitivity is the most important for widespread application. Nonetheless, MFC-based biosensors represent a promising approach towards single pollutant detection.

BIOSENSORS FOR ENVIRONMENTAL APPLICATIONS

EPA Science Inventory

A review, with 19 references, is given on challenges and possible opportunities for the development of biosensors for environmental monitoring applications. The high cost and slow turnaround times typically associated with the measurement of regulated pollutants clearly indicates...

Application of genetically engineered microbial whole-cell biosensors for combined chemosensing.

PubMed

He, Wei; Yuan, Sheng; Zhong, Wen-Hui; Siddikee, Md Ashaduzzaman; Dai, Chuan-Chao

2016-02-01

The progress of genetically engineered microbial whole-cell biosensors for chemosensing and monitoring has been developed in the last 20 years. Those biosensors respond to target chemicals and produce output signals, which offer a simple and alternative way of assessment approaches. As actual pollution caused by human activities usually contains a combination of different chemical substances, how to employ those biosensors to accurately detect real contaminant samples and evaluate biological effects of the combined chemicals has become a realistic object of environmental researches. In this review, we outlined different types of the recent method of genetically engineered microbial whole-cell biosensors for combined chemical evaluation, epitomized their detection performance, threshold, specificity, and application progress that have been achieved up to now. We also discussed the applicability and limitations of this biosensor technology and analyzed the optimum conditions for their environmental assessment in a combined way.

A reduced graphene oxide based electrochemical biosensor for tyrosine detection

NASA Astrophysics Data System (ADS)

Wei, Junhua; Qiu, Jingjing; Li, Li; Ren, Liqiang; Zhang, Xianwen; Chaudhuri, Jharna; Wang, Shiren

2012-08-01

In this paper, a ‘green’ and safe hydrothermal method has been used to reduce graphene oxide and produce hemin modified graphene nanosheet (HGN) based electrochemical biosensors for the determination of l-tyrosine levels. The as-fabricated HGN biosensors were characterized by UV-visible absorption spectra, fluorescence spectra, Fourier transform infrared spectroscopy (FTIR) spectra and thermogravimetric analysis (TGA). The experimental results indicated that hemin was successfully immobilized on the reduced graphene oxide nanosheet (rGO) through π-π interaction. TEM images and EDX results further confirmed the attachment of hemin on the rGO nanosheet. Cyclic voltammetry tests were carried out for the bare glass carbon electrode (GCE), the rGO electrode (rGO/GCE), and the hemin-rGO electrode (HGN/GCE). The HGN/GCE based biosensor exhibits a tyrosine detection linear range from 5 × 10-7 M to 2 × 10-5 M with a detection limitation of 7.5 × 10-8 M at a signal-to-noise ratio of 3. The sensitivity of this biosensor is 133 times higher than that of the bare GCE. In comparison with other works, electroactive biosensors are easily fabricated, easily controlled and cost-effective. Moreover, the hemin-rGO based biosensors demonstrate higher stability, a broader detection linear range and better detection sensitivity. Study of the oxidation scheme reveals that the rGO enhances the electron transfer between the electrode and the hemin, and the existence of hemin groups effectively electrocatalyzes the oxidation of tyrosine. This study contributes to a widespread clinical application of nanomaterial based biosensor devices with a broader detection linear range, improved stability, enhanced sensitivity and reduced costs.

Development of phage/antibody immobilized magnetostrictive biosensors

NASA Astrophysics Data System (ADS)

Fu, Liling

There is an urgent need for biosensors that are able to detect and quantify the presence of a small amount of pathogens in a real-time manner accurately and quickly to guide prevention efforts and assay food and water quality. Acoustic wave (AW) devices, whose performance is defined by mass sensitivity (Sm) and quality factor (Q value), have been extensively studied as high performance biosensor platforms. However, current AW devices still face some challenges such as the difficulty to be employed in liquid and low Q value in practical applications. The objective of this research is to develop magnetostrictive sensors which include milli/microcantilever type (MSMC) and particle type (MSP). Compared to other AW devices, MSMC exhibits the following advantages: (1) wireless/remote driving and sensing; (2) easy to fabricate; (3) works well in liquid; (4) exhibits a high Q value (> 500 in air). The fundamental study of the damping effect on MSMCs from the surrounding media including air and liquids were conducted to improve the Q value of MSMCs. The experiment results show that the Q value is dependent on the properties of surrounding media (e.g. viscosity, density), the geometry of the MSMCs, and the harmonic mode on the resonance behavior of MSMCs, etc. The phage-coated MSMC has high specificity and sensitivity even while used in water with a low concentration of targeted bacteria. Two currently developed phages, JRB7 and E2, respectively respond to Bacillus anthracis spores and Salmonella typhimurium, were employed as bio-recognition elements in this research. The phage-immobilized MSMC biosensors exhibited high performance and detection of limit was 5 x 104 cfu/ml for the MSMC in size of 1.4 x 0.8 x 0.035 mm. The MSMC-based biosensors were indicated as a very potential method for in-situ monitoring of the biological quality in water. The MSP combine antibody was used to detect Staphylococcus aureus in this experiment. The interface between MSPs and antibody was

Nano-particle enhanced impedimetric biosensor for detedtion of foodborne pathogens

NASA Astrophysics Data System (ADS)

Kim, G.; Om, A. S.; Mun, J. H.

2007-03-01

Recent outbreaks of foodborne illness have been increased the need for rapid and sensitive methods for detection of these pathogens. Conventional methods for pathogens detection and identification involve prolonged multiple enrichment steps. Even though some immunological rapid assays are available, these assays still need enrichment steps result in delayed detection. Biosensors have shown great potential for rapid detection of foodborne pathogens. They are capable of direct monitoring the antigen-antibody reactions in real time. Among the biosensors, impedimetric biosensors have been widely adapted as an analysis tool for the study of various biological binding reactions because of their high sensitivity and reagentless operation. In this study a nanoparticle-enhanced impedimetric biosensor for Salmonella enteritidis detection was developed which detected impedance changes caused by the attachment of the cells to the anti-Salmonella antibodies immobilized on interdigitated gold electrodes. Successive immobilization of neutravidin followed by anti-Salmonella antibodies was performed to the sensing area to create a biological detection surface. To enhance the impedance responses generated by antigen-antibody reactions, anti-Salmonella antibody conjugated nanoparticles were introduced on the sensing area. Using a portable impedance analyzer, the impedance across the interdigital electrodes was measured after the series of antigen-antibody bindings. Bacteria cells present in solution attached to capture antibodies and became tethered to the sensor surface. Attached bacteria cells changed the dielectric constant of the media between the electrodes thereby causing a change in measured impedance. Optimum input frequency was determined by analyzing frequency characteristics of the biosensor over ranges of applied frequencies from 10 Hz to 400 Hz. At 100 Hz of input frequency, the biosensor was most sensitive to the changes of the bacteria concentration and this frequency

Nanogravimetric and voltammetric DNA-hybridization biosensors for studies of DNA damage by common toxicants and pollutants.

PubMed

Nowicka, Anna M; Kowalczyk, Agata; Stojek, Zbigniew; Hepel, Maria

2010-01-01

Electrochemical and nanogravimetric DNA-hybridization biosensors have been developed for sensing single mismatches in the probe-target ssDNA sequences. The voltammetric transduction was achieved by coupling ferrocene moiety to streptavidin linked to biotinylated tDNA. The mass-related frequency transduction was implemented by immobilizing the sensory pDNA on a gold-coated quartz crystal piezoresonators oscillating in the 10MHz band. The high sensitivity of these sensors enabled us to study DNA damage caused by representative toxicants and environmental pollutants, including Cr(VI) species, common pesticides and herbicides. We have found that the sensor responds rapidly to any damage caused by Cr(VI) species, with more severe DNA damage observed for Cr(2)O(7)(2-) and for CrO(4)(2-) in the presence of H(2)O(2) as compared to CrO(4)(2-) alone. All herbicides and pesticides examined caused DNA damage or structural alterations leading to the double-helix unwinding. Among these compounds, paraoxon-ethyl and atrazine caused the fastest and most severe damage to DNA. The physico-chemical mechanism of damaging interactions between toxicants and DNA has been proposed. The methodology of testing voltammetric and nanogravimetric DNA-hybridization biosensors developed in this work can be employed as a simple protocol to obtain rapid comparative data concerning DNA damage caused by herbicide, pesticides and other toxic pollutants. The DNA-hybridization biosensor can, therefore, be utilized as a rapid screening device for classifying environmental pollutants and to evaluate DNA damage induced by these compounds.

Green Chemistry Glucose Biosensor Development using Etlingera elatior Extract

NASA Astrophysics Data System (ADS)

Fatoni, A.; Anggraeni, M. D.; Zusfahair; Iqlima, H.

2018-01-01

Glucose biosensor development is one of the important strategies for early detection of diabetes mellitus disease. This study was aimed to explore the flower extract of Etlingera elatior for a green-analysis method of glucose biosensor. Flowers were extracted using ethanol: HCl and tested its performances as an indicator of glucose biosensor using glucose oxidase enzyme. The glucose oxidase react with glucose resulted hydrogen peroxide that would change the color of the flower extract. Furthermore, the extract was also studied including their stability to pH, oxidizing and reducing, temperature, and storage. The results showed that the Etlingera elatior extract had high correlation between color change and glucose concentration with regression equation of y = -0.0005x + 0.4724 and R2 of 0.9965. The studied biosensor showed a wide linear range to detect glucose sample of 0 to 500 mM. The extract characterization showed a more stable in low pH (acid), reducing agent addition, heating treatment and storage.

Engineering prokaryotic transcriptional activators as metabolite biosensors in yeast.

PubMed

Skjoedt, Mette L; Snoek, Tim; Kildegaard, Kanchana R; Arsovska, Dushica; Eichenberger, Michael; Goedecke, Tobias J; Rajkumar, Arun S; Zhang, Jie; Kristensen, Mette; Lehka, Beata J; Siedler, Solvej; Borodina, Irina; Jensen, Michael K; Keasling, Jay D

2016-11-01

Whole-cell biocatalysts have proven a tractable path toward sustainable production of bulk and fine chemicals. Yet the screening of libraries of cellular designs to identify best-performing biocatalysts is most often a low-throughput endeavor. For this reason, the development of biosensors enabling real-time monitoring of production has attracted attention. Here we applied systematic engineering of multiple parameters to search for a general biosensor design in the budding yeast Saccharomyces cerevisiae based on small-molecule binding transcriptional activators from the prokaryote superfamily of LysR-type transcriptional regulators (LTTRs). We identified a design supporting LTTR-dependent activation of reporter gene expression in the presence of cognate small-molecule inducers. As proof of principle, we applied the biosensors for in vivo screening of cells producing naringenin or cis,cis-muconic acid at different levels, and found that reporter gene output correlated with production. The transplantation of prokaryotic transcriptional activators into the eukaryotic chassis illustrates the potential of a hitherto untapped biosensor resource useful for biotechnological applications.

Electrochemical Enzyme Biosensors Revisited: Old Solutions for New Problems.

PubMed

Monteiro, Tiago; Almeida, Maria Gabriela

2018-05-14

Worldwide legislation is driving the development of novel and highly efficient analytical tools for assessing the composition of every material that interacts with Consumers or Nature. The biosensor technology is one of the most active R&D domains of Analytical Sciences focused on the challenge of taking analytical chemistry to the field. Electrochemical biosensors based on redox enzymes, in particular, are highly appealing due to their usual quick response, high selectivity and sensitivity, low cost and portable dimensions. This review paper aims to provide an overview of the most important advances made in the field since the proposal of the first biosensor, the well-known hand-held glucose meter. The first section addresses the current needs and challenges for novel analytical tools, followed by a brief description of the different components and configurations of biosensing devices, and the fundamentals of enzyme kinetics and amperometry. The following sections emphasize on enzyme-based amperometric biosensors and the different stages of their development.

Paper electrodes for bioelectrochemistry: Biosensors and biofuel cells.

PubMed

Desmet, Cloé; Marquette, Christophe A; Blum, Loïc J; Doumèche, Bastien

2016-02-15

Paper-based analytical devices (PAD) emerge in the scientific community since 2007 as low-cost, wearable and disposable devices for point-of-care diagnostic due to the widespread availability, long-time knowledge and easy manufacturing of cellulose. Rapidly, electrodes were introduced in PAD for electrochemical measurements. Together with biological components, a new generation of electrochemical biosensors was born. This review aims to take an inventory of existing electrochemical paper-based biosensors and biofuel cells and to identify, at the light of newly acquired data, suitable methodologies and crucial parameters in this field. Paper selection, electrode material, hydrophobization of cellulose, dedicated electrochemical devices and electrode configuration in biosensors and biofuel cells will be discussed. Copyright © 2015 Elsevier B.V. All rights reserved.

Potentiometric Urea Biosensor Based on an Immobilised Fullerene-Urease Bio-Conjugate

PubMed Central

Saeedfar, Kasra; Heng, Lee Yook; Ling, Tan Ling; Rezayi, Majid

2013-01-01

A novel method for the rapid modification of fullerene for subsequent enzyme attachment to create a potentiometric biosensor is presented. Urease was immobilized onto the modified fullerene nanomaterial. The modified fullerene-immobilized urease (C60-urease) bioconjugate has been confirmed to catalyze the hydrolysis of urea in solution. The biomaterial was then deposited on a screen-printed electrode containing a non-plasticized poly(n-butyl acrylate) (PnBA) membrane entrapped with a hydrogen ionophore. This pH-selective membrane is intended to function as a potentiometric urea biosensor with the deposition of C60-urease on the PnBA membrane. Various parameters for fullerene modification and urease immobilization were investigated. The optimal pH and concentration of the phosphate buffer for the urea biosensor were 7.0 and 0.5 mM, respectively. The linear response range of the biosensor was from 2.31 × 10−3 M to 8.28 × 10−5 M. The biosensor's sensitivity was 59.67 ± 0.91 mV/decade, which is close to the theoretical value. Common cations such as Na+, K+, Ca2+, Mg2+ and NH4+ showed no obvious interference with the urea biosensor's response. The use of a fullerene-urease bio-conjugate and an acrylic membrane with good adhesion prevented the leaching of urease enzyme and thus increased the stability of the urea biosensor for up to 140 days. PMID:24322561

Development of mediator-type biosensor to wirelessly monitor whole cholesterol concentration in fish.

PubMed

Takase, Mai; Murata, Masataka; Hibi, Kyoko; Huifeng, Ren; Endo, Hideaki

2014-04-01

We developed a wireless monitoring system to monitor fish condition by tracking the change in whole cholesterol concentration. The whole cholesterol concentration of fish is a source of steroid hormones or indicator of immunity level, which makes its detection important for tracking physiological condition of fish. Wireless monitoring system comprises of mediator-type biosensor and wireless transmission device. Biosensor is implantable to fish body, and transmission device is so light, in that fish is allowed to swim freely during monitoring. Cholesterol esterase and oxidase were fixated on to the detection site of biosensor and used to detect the whole cholesterol concentration. However, cholesterol oxidase incorporates oxidation-reduction reaction of oxygen for detection, which concentration fluctuates easily due to change in environmental condition. Meanwhile, mediator-type biosensor enables monitoring of whole cholesterol concentration by using mediator to substitute that oxidation-reduction reaction of oxygen. Characteristic of fabricated mediator-type biosensor was tested. The sensor output current of mediator-type biosensor remained stable compared to output current of non-mediator-type biosensor under fluctuating oxygen concentration of 0-8 ppm, which implied that this sensor is less affected by change in dissolved oxygen concentration. That biosensor was then implanted into fish for wireless monitoring. As a result, approximately 48 h of real-time monitoring was successful.

Design and application of a lactulose biosensor.

PubMed

Wu, Jieyuan; Jiang, Peixia; Chen, Wei; Xiong, Dandan; Huang, Linglan; Jia, Junying; Chen, Yuanyuan; Jin, Jian-Ming; Tang, Shuang-Yan

2017-04-07

In this study the repressor of Escherichia coli lac operon, LacI, has been engineered for altered effector specificity. A LacI saturation mutagenesis library was subjected to Fluorescence Activated Cell Sorting (FACS) dual screening. Mutant LacI-L5 was selected and it is specifically induced by lactulose but not by other disaccharides tested (lactose, epilactose, maltose, sucrose, cellobiose and melibiose). LacI-L5 has been successfully used to construct a whole-cell lactulose biosensor which was then applied in directed evolution of cellobiose 2-epimerase (C2E) for elevated lactulose production. The mutant C2E enzyme with ~32-fold enhanced expression level was selected, demonstrating the high efficiency of the lactulose biosensor. LacI-L5 can also be used as a novel regulatory tool. This work explores the potential of engineering LacI for customized molecular biosensors which can be applied in practice.

Thin Hydrogel Films for Optical Biosensor Applications

PubMed Central

Mateescu, Anca; Wang, Yi; Dostalek, Jakub; Jonas, Ulrich

2012-01-01

Hydrogel materials consisting of water-swollen polymer networks exhibit a large number of specific properties highly attractive for a variety of optical biosensor applications. This properties profile embraces the aqueous swelling medium as the basis of biocompatibility, non-fouling behavior, and being not cell toxic, while providing high optical quality and transparency. The present review focuses on some of the most interesting aspects of surface-attached hydrogel films as active binding matrices in optical biosensors based on surface plasmon resonance and optical waveguide mode spectroscopy. In particular, the chemical nature, specific properties, and applications of such hydrogel surface architectures for highly sensitive affinity biosensors based on evanescent wave optics are discussed. The specific class of responsive hydrogel systems, which can change their physical state in response to externally applied stimuli, have found large interest as sophisticated materials that provide a complex behavior to hydrogel-based sensing devices. PMID:24957962

Xanthine oxidase biosensor for monitoring meat spoilage

NASA Astrophysics Data System (ADS)

Vanegas, D. C.; Gomes, C.; McLamore, E. S.

2014-05-01

In this study, we have designed an electrochemical biosensor for real-time detection of specific biomarkers of bacterial metabolism related to meat spoilage (hypoxanthine and xanthine). The selective biosensor was developed by assembling a `sandwich' of nanomaterials and enzymes on a platinum-iridium electrode (1.6 mm tip diameter). The materials deposited on the sensor tip include amorphous platinum nanoclusters (i.e. Pt black), reduced graphene oxide, nanoceria, and xanthine oxidase. Xanthine oxidase was encapsulated in laponite hydrogel and used for the biorecognition of hypoxanthine and xanthine (two molecules involved in the rotting of meat by spoilage microorganisms). The developed biosensor demonstrated good electrochemical performance toward xanthine with sensitivity of 2.14 +/- 1.48 μA/mM, response time of 5.2 +/- 1.5 sec, lower detection limit of 150 +/- 39 nM, and retained at least 88% of its activity after 7 days of continuous use.

Microplate technique for determining accumulation of metals by algae

DOE Office of Scientific and Technical Information (OSTI.GOV)

Hassett, J.M.; Jennett, J.C.; Smith, J.E.

1981-05-01

A microplate technique was developed to determine the conditions under which pure cultures of algae removed heavy metals from aqueous solutions. Variables investigated included algal species and strain, culture age (11 and 44 days), metal (mercury, lead, cadmium, and zinc), pH, effects of different buffer solutions, and time of exposure. Plastic, U-bottomed microtiter plates were used in conjunction with heavy metal radionuclides to determine concentration factors for metal-alga combinations. The technique developed was rapid, statistically reliable, and economical of materials and cells. All species of algae studied removed mercury from solution. Green algae proved better at accumulating cadmium than didmore » blue-green algae. No alga studied removed zinc, perhaps because cells were maintained in the dark during the labeling period. Chlamydomonas sp. proved superior in ability to remove lead from solution.« less

Sensitive detection of maltose and glucose based on dual enzyme-displayed bacteria electrochemical biosensor.

PubMed

Liu, Aihua; Lang, Qiaolin; Liang, Bo; Shi, Jianguo

2017-01-15

Glucoamylase-displayed bacteria (GA-bacteria) and glucose dehydrogenase-displayed bacteria (GDH-bacteria) were co-immobilized on multi-walled carbon nanotubes (MWNTs) modified glassy carbon electrode (GCE) to construct GA-bacteria/GDH-bacteria/MWNTs/GCE biosensor. The biosensor was developed by optimizing the loading amount and the ratio of GA-bacteria to GDH-bacteria. The as-prepared biosensor exhibited a wide dynamic range of 0.2-10mM and a low detection limit of 0.1mM maltose (S/N=3). The biosensor also had a linear response to glucose in the range of 0.1-2.0mM and a low detection limit of 0.04mM glucose (S/N=3). Interestingly, at the same concentration, glucose was 3.75-fold sensitive than that of maltose at the proposed biosensor. No interferences were observed for other possible mono- and disaccharides. The biosensor also demonstrated good long-term storage stability and repeatability. Further, using both GDH-bacteria/MWNTs/GCE biosensor and GA-bacteria/GDH-bacteria/MWNTs/GCE biosensor, glucose and maltose in real samples can be detected. Therefore, the proposed biosensor is capable of monitoring the food manufacturing and fermentation process. Copyright © 2016 Elsevier B.V. All rights reserved.

Nanomolar detection of methylparaben by a cost-effective hemoglobin-based biosensor.

PubMed

Hajian, A; Ghodsi, J; Afraz, A; Yurchenko, O; Urban, G

2016-12-01

This work describes the development of a new biosensor for methylparaben determination using electrocatalytic properties of hemoglobin in the presence of hydrogen peroxide. The voltammetric oxidation of methylparaben by the proposed biosensor in phosphate buffer (pH=7.0), a physiological pH, was studied and it was confirmed that methylparaben undergoes a one electron-one proton reaction in a diffusion-controlled process. The biosensor was fabricated by carbon paste electrode modified with hemoglobin and multiwalled carbon nanotube. Based on the excellent electrochemical properties of the modified electrode, a sensitive voltammetric method was used for determination of methylparaben within a linear range from 0.1 to 13μmolL(-1) and detection limit of 25nmolL(-1). The developed biosensor possessed accurate and rapid response to methylparaben and showed good sensitivity, stability, and repeatability. Finally, the applicability of the proposed biosensor was verified by methylparaben evaluation in various real samples. Copyright © 2016 Elsevier B.V. All rights reserved.

Amine oxidase-based biosensors for spermine and spermidine determination.

PubMed

Boffi, Alberto; Favero, Gabriele; Federico, Rodolfo; Macone, Alberto; Antiochia, Riccarda; Tortolini, Cristina; Sanzó, Gabriella; Mazzei, Franco

2015-02-01

The present work describes the development and optimization of electrochemical biosensors for specific determination of the biogenic polyamine spermine (Spm) and spermidine (Spmd) whose assessment represents a novel important analytical tool in food analysis and human diagnostics. These biosensors have been prepared using novel engineered enzymes: polyamine oxidase (PAO) endowed with selectivity towards Spm and Spmd and spermine oxidase (SMO) characterized by strict specificity towards Spm. The current design entails biosensors in which the enzymes were entrapped in poly(vinyl alcohol) bearing styrylpyridinium groups (PVA-SbQ), a photocrosslinkable gel, onto an electrode surface. Screen-printed electrodes (SPEs) were used as electrochemical transducers for enzymatically produced hydrogen peroxide, operating at different potential vs Ag/AgCl according to the material of the working electrode (WE): +700 mV for graphite (GP) or -100 mV for Prussian blue (PB)-modified SPE, respectively. Biosensor performances were evaluated by means of flow injection amperometric (FIA) measurements. The modified electrodes showed good sensitivity, long-term stability and reproducibility. Under optimal conditions, the PAO biosensor showed a linear range 0.003-0.3 mM for Spm and 0.01-0.4 mM for Spmd, while with the SMO biosensor, a linear range of 0.004-0.5 mM for Spm has been obtained. The main kinetic parameters apparent Michaelis constant (K M), turnover number (K cat) and steady-state current (I max) were determined. The proposed device was then applied to the determination of biogenic amines in blood samples. The results obtained were in good agreement with those obtained with the GC-MS reference method.

A global benchmark study using affinity-based biosensors

PubMed Central

Rich, Rebecca L.; Papalia, Giuseppe A.; Flynn, Peter J.; Furneisen, Jamie; Quinn, John; Klein, Joshua S.; Katsamba, Phini S.; Waddell, M. Brent; Scott, Michael; Thompson, Joshua; Berlier, Judie; Corry, Schuyler; Baltzinger, Mireille; Zeder-Lutz, Gabrielle; Schoenemann, Andreas; Clabbers, Anca; Wieckowski, Sebastien; Murphy, Mary M.; Page, Phillip; Ryan, Thomas E.; Duffner, Jay; Ganguly, Tanmoy; Corbin, John; Gautam, Satyen; Anderluh, Gregor; Bavdek, Andrej; Reichmann, Dana; Yadav, Satya P.; Hommema, Eric; Pol, Ewa; Drake, Andrew; Klakamp, Scott; Chapman, Trevor; Kernaghan, Dawn; Miller, Ken; Schuman, Jason; Lindquist, Kevin; Herlihy, Kara; Murphy, Michael B.; Bohnsack, Richard; Andrien, Bruce; Brandani, Pietro; Terwey, Danny; Millican, Rohn; Darling, Ryan J.; Wang, Liann; Carter, Quincy; Dotzlaf, Joe; Lopez-Sagaseta, Jacinto; Campbell, Islay; Torreri, Paola; Hoos, Sylviane; England, Patrick; Liu, Yang; Abdiche, Yasmina; Malashock, Daniel; Pinkerton, Alanna; Wong, Melanie; Lafer, Eileen; Hinck, Cynthia; Thompson, Kevin; Primo, Carmelo Di; Joyce, Alison; Brooks, Jonathan; Torta, Federico; Bagge Hagel, Anne Birgitte; Krarup, Janus; Pass, Jesper; Ferreira, Monica; Shikov, Sergei; Mikolajczyk, Malgorzata; Abe, Yuki; Barbato, Gaetano; Giannetti, Anthony M.; Krishnamoorthy, Ganeshram; Beusink, Bianca; Satpaev, Daulet; Tsang, Tiffany; Fang, Eric; Partridge, James; Brohawn, Stephen; Horn, James; Pritsch, Otto; Obal, Gonzalo; Nilapwar, Sanjay; Busby, Ben; Gutierrez-Sanchez, Gerardo; Gupta, Ruchira Das; Canepa, Sylvie; Witte, Krista; Nikolovska-Coleska, Zaneta; Cho, Yun Hee; D’Agata, Roberta; Schlick, Kristian; Calvert, Rosy; Munoz, Eva M.; Hernaiz, Maria Jose; Bravman, Tsafir; Dines, Monica; Yang, Min-Hsiang; Puskas, Agnes; Boni, Erica; Li, Jiejin; Wear, Martin; Grinberg, Asya; Baardsnes, Jason; Dolezal, Olan; Gainey, Melicia; Anderson, Henrik; Peng, Jinlin; Lewis, Mark; Spies, Peter; Trinh, Quyhn; Bibikov, Sergei; Raymond, Jill; Yousef, Mohammed; Chandrasekaran, Vidya; Feng, Yuguo; Emerick, Anne; Mundodo, Suparna; Guimaraes, Rejane; McGirr, Katy; Li, Yue-Ji; Hughes, Heather; Mantz, Hubert; Skrabana, Rostislav; Witmer, Mark; Ballard, Joshua; Martin, Loic; Skladal, Petr; Korza, George; Laird-Offringa, Ite; Lee, Charlene S.; Khadir, Abdelkrim; Podlaski, Frank; Neuner, Phillippe; Rothacker, Julie; Rafique, Ashique; Dankbar, Nico; Kainz, Peter; Gedig, Erk; Vuyisich, Momchilo; Boozer, Christina; Ly, Nguyen; Toews, Mark; Uren, Aykut; Kalyuzhniy, Oleksandr; Lewis, Kenneth; Chomey, Eugene; Pak, Brian J.; Myszka, David G.

2013-01-01

To explore the variability in biosensor studies, 150 participants from 20 countries were given the same protein samples and asked to determine kinetic rate constants for the interaction. We chose a protein system that was amenable to analysis using different biosensor platforms as well as by users of different expertise levels. The two proteins (a 50-kDa Fab and a 60-kDa glutathione S-transferase [GST] antigen) form a relatively high-affinity complex, so participants needed to optimize several experimental parameters, including ligand immobilization and regeneration conditions as well as analyte concentrations and injection/dissociation times. Although most participants collected binding responses that could be fit to yield kinetic parameters, the quality of a few data sets could have been improved by optimizing the assay design. Once these outliers were removed, the average reported affinity across the remaining panel of participants was 620 pM with a standard deviation of 980 pM. These results demonstrate that when this biosensor assay was designed and executed appropriately, the reported rate constants were consistent, and independent of which protein was immobilized and which biosensor was used. PMID:19133223

Biomolecular logic systems: applications to biosensors and bioactuators

NASA Astrophysics Data System (ADS)

Katz, Evgeny

2014-05-01

The paper presents an overview of recent advances in biosensors and bioactuators based on the biocomputing concept. Novel biosensors digitally process multiple biochemical signals through Boolean logic networks of coupled biomolecular reactions and produce output in the form of YES/NO response. Compared to traditional single-analyte sensing devices, biocomputing approach enables a high-fidelity multi-analyte biosensing, particularly beneficial for biomedical applications. Multi-signal digital biosensors thus promise advances in rapid diagnosis and treatment of diseases by processing complex patterns of physiological biomarkers. Specifically, they can provide timely detection and alert to medical emergencies, along with an immediate therapeutic intervention. Application of the biocomputing concept has been successfully demonstrated for systems performing logic analysis of biomarkers corresponding to different injuries, particularly exemplified for liver injury. Wide-ranging applications of multi-analyte digital biosensors in medicine, environmental monitoring and homeland security are anticipated. "Smart" bioactuators, for example for signal-triggered drug release, were designed by interfacing switchable electrodes and biocomputing systems. Integration of novel biosensing and bioactuating systems with the biomolecular information processing systems keeps promise for further scientific advances and numerous practical applications.

Tyrosinase-Based Biosensors for Selective Dopamine Detection

PubMed Central

Florescu, Monica; David, Melinda

2017-01-01

A novel tyrosinase-based biosensor was developed for the detection of dopamine (DA). For increased selectivity, gold electrodes were previously modified with cobalt (II)-porphyrin (CoP) film with electrocatalytic activity, to act both as an electrochemical mediator and an enzyme support, upon which the enzyme tyrosinase (Tyr) was cross-linked. Differential pulse voltammetry was used for electrochemical detection and the reduction current of dopamine-quinone was measured as a function of dopamine concentration. Our experiments demonstrated that the presence of CoP improves the selectivity of the electrode towards dopamine in the presence of ascorbic acid (AA), with a linear trend of concentration dependence in the range of 2–30 µM. By optimizing the conditioning parameters, a separation of 130 mV between the peak potentials for ascorbic acid AA and DA was obtained, allowing the selective detection of DA. The biosensor had a sensitivity of 1.22 ± 0.02 µA·cm−2·µM−1 and a detection limit of 0.43 µM. Biosensor performances were tested in the presence of dopamine medication, with satisfactory results in terms of recovery (96%), and relative standard deviation values below 5%. These results confirmed the applicability of the biosensors in real samples such as human urine and blood serum. PMID:28590453

Label-free electrical detection using carbon nanotube-based biosensors.

PubMed

Maehashi, Kenzo; Matsumoto, Kazuhiko

2009-01-01

Label-free detections of biomolecules have attracted great attention in a lot of life science fields such as genomics, clinical diagnosis and practical pharmacy. In this article, we reviewed amperometric and potentiometric biosensors based on carbon nanotubes (CNTs). In amperometric detections, CNT-modified electrodes were used as working electrodes to significantly enhance electroactive surface area. In contrast, the potentiometric biosensors were based on aptamer-modified CNT field-effect transistors (CNTFETs). Since aptamers are artificial oligonucleotides and thus are smaller than the Debye length, proteins can be detected with high sensitivity. In this review, we discussed on the technology, characteristics and developments for commercialization in label-free CNT-based biosensors.

A reduced graphene oxide based electrochemical biosensor for tyrosine detection.

PubMed

Wei, Junhua; Qiu, Jingjing; Li, Li; Ren, Liqiang; Zhang, Xianwen; Chaudhuri, Jharna; Wang, Shiren

2012-08-24

In this paper, a 'green' and safe hydrothermal method has been used to reduce graphene oxide and produce hemin modified graphene nanosheet (HGN) based electrochemical biosensors for the determination of l-tyrosine levels. The as-fabricated HGN biosensors were characterized by UV-visible absorption spectra, fluorescence spectra, Fourier transform infrared spectroscopy (FTIR) spectra and thermogravimetric analysis (TGA). The experimental results indicated that hemin was successfully immobilized on the reduced graphene oxide nanosheet (rGO) through π-π interaction. TEM images and EDX results further confirmed the attachment of hemin on the rGO nanosheet. Cyclic voltammetry tests were carried out for the bare glass carbon electrode (GCE), the rGO electrode (rGO/GCE), and the hemin-rGO electrode (HGN/GCE). The HGN/GCE based biosensor exhibits a tyrosine detection linear range from 5 × 10(-7) M to 2 × 10(-5) M with a detection limitation of 7.5 × 10(-8) M at a signal-to-noise ratio of 3. The sensitivity of this biosensor is 133 times higher than that of the bare GCE. In comparison with other works, electroactive biosensors are easily fabricated, easily controlled and cost-effective. Moreover, the hemin-rGO based biosensors demonstrate higher stability, a broader detection linear range and better detection sensitivity. Study of the oxidation scheme reveals that the rGO enhances the electron transfer between the electrode and the hemin, and the existence of hemin groups effectively electrocatalyzes the oxidation of tyrosine. This study contributes to a widespread clinical application of nanomaterial based biosensor devices with a broader detection linear range, improved stability, enhanced sensitivity and reduced costs.

Battlefield Medical Network: Biosensors In A Tactical Environment

DTIC Science & Technology

2016-03-01

MEDICAL NETWORK: BIOSENSORS IN A TACTICAL ENVIRONMENT by Ralph R. Montgomery Yekaterina L. Anderson March 2016 Thesis Advisor: Alex...2016 3. REPORT TYPE AND DATES COVERED Master’s thesis 4. TITLE AND SUBTITLE BATTLEFIELD MEDICAL NETWORK: BIOSENSORS IN A TACTICAL ENVIRONMENT 5...School Monterey, CA 93943-5000 8. PERFORMING ORGANIZATION REPORT NUMBER 9. SPONSORING /MONITORING AGENCY NAME(S) AND ADDRESS(ES) N/ A 10

Mechanistic Challenges and Advantages of Biosensor Miniaturization into the Nanoscale.

PubMed

Soleymani, Leyla; Li, Feng

2017-04-28

Over the past few decades, there has been tremendous interest in developing biosensing systems that combine high sensitivity and specificity with rapid sample-to-answer times, portability, low-cost operation, and ease-of-use. Miniaturizing the biosensor dimensions into the nanoscale has been identified as a strategy for addressing the functional requirements of point-of-care and wearable biosensors. However, it is important to consider that decreasing the critical dimensions of biosensing elements impacts the two most important performance metrics of biosensors: limit-of-detection and response time. Miniaturization into the nanoscale enhances signal-to-noise-ratio by increasing the signal density (signal/geometric surface area) and reducing background signals. However, there is a trade-off between the enhanced signal transduction efficiency and the longer time it takes to collect target analytes on sensor surfaces due to the increase in mass transport times. By carefully considering the signal transduction mechanisms and reaction-transport kinetics governing different classes of biosensors, it is possible to develop structure-level and device-level strategies for leveraging miniaturization toward creating biosensors that combine low limit-of-detection with rapid response times.

Last Advances in Silicon-Based Optical Biosensors.

PubMed

Fernández Gavela, Adrián; Grajales García, Daniel; Ramirez, Jhonattan C; Lechuga, Laura M

2016-02-24

We review the most important achievements published in the last five years in the field of silicon-based optical biosensors. We focus specially on label-free optical biosensors and their implementation into lab-on-a-chip platforms, with an emphasis on developments demonstrating the capability of the devices for real bioanalytical applications. We report on novel transducers and materials, improvements of existing transducers, new and improved biofunctionalization procedures as well as the prospects for near future commercialization of these technologies.

Last Advances in Silicon-Based Optical Biosensors

PubMed Central

Fernández Gavela, Adrián; Grajales García, Daniel; Ramirez, Jhonattan C.; Lechuga, Laura M.

2016-01-01

We review the most important achievements published in the last five years in the field of silicon-based optical biosensors. We focus specially on label-free optical biosensors and their implementation into lab-on-a-chip platforms, with an emphasis on developments demonstrating the capability of the devices for real bioanalytical applications. We report on novel transducers and materials, improvements of existing transducers, new and improved biofunctionalization procedures as well as the prospects for near future commercialization of these technologies. PMID:26927105

A protocatechuate biosensor for Pseudomonas putida KT2440 via promoter and protein evolution.

PubMed

Jha, Ramesh K; Bingen, Jeremy M; Johnson, Christopher W; Kern, Theresa L; Khanna, Payal; Trettel, Daniel S; Strauss, Charlie E M; Beckham, Gregg T; Dale, Taraka

2018-06-01

Robust fluorescence-based biosensors are emerging as critical tools for high-throughput strain improvement in synthetic biology. Many biosensors are developed in model organisms where sophisticated synthetic biology tools are also well established. However, industrial biochemical production often employs microbes with phenotypes that are advantageous for a target process, and biosensors may fail to directly transition outside the host in which they are developed. In particular, losses in sensitivity and dynamic range of sensing often occur, limiting the application of a biosensor across hosts. Here we demonstrate the optimization of an Escherichia coli- based biosensor in a robust microbial strain for the catabolism of aromatic compounds, Pseudomonas putida KT2440, through a generalizable approach of modulating interactions at the protein-DNA interface in the promoter and the protein-protein dimer interface. The high-throughput biosensor optimization approach demonstrated here is readily applicable towards other allosteric regulators.

Construction of uric acid biosensor based on biomimetic titanate nanotubes.

PubMed

Tao, Haisheng; Wang, Xuebin; Wang, Xizhang; Hu, Yemin; Ma, Yanwen; Lu, Yinong; Hu, Zheng

2010-02-01

A uric acid biosensor has been fabricated through the immobilization of uricase on glassy carbon electrode modified by biomimetic titanate nanotubes of high specific surface area synthesized by hydrothermal decomposition. The so-constructed biosensor presents a high affinity to uric acid with a small apparent Michaelis-Menten constant of only 0.66 mM. The biosensor exhibits fairly good electrochemical properties such as the high sensitivity of 184.3 microAcm(-2)mM(-1), the fast response of less than 2 s, as well as the wide linear range from 1 microM to 5 mM. These performances indicate that titanate nanotubes could provide a favorable microenvironment for uricase immobilization, stabilize its biological activity, and function as an efficient electron conducting tunnel to facilitate the electron transfer. This suggests an important potential of titanate nanotubes in uric acid biosensors.

Rapid detection of Salmonella Typhimurium in chicken carcass using a SPR biosensor

NASA Astrophysics Data System (ADS)

Wang, Shizhou; Lan, Yubin; Yin, Yongguang; Dasari, Thirumala R.

2005-11-01

The SPR biosensor was sensitive to the presence of Salmonella Typhimurium in chicken carcass. The selectivity of the SPR biosensor was assayed using a series of antibody concentrations and dilution series of the organism. The SPR biosensor was specific to Salmonella Typhimurium at concentrations of 106 CFU/ml. Initial results show potential for its application for pathogenic bacteria monitoring.

A glucose biosensor based on partially unzipped carbon nanotubes.

PubMed

Hu, Huifang; Feng, Miao; Zhan, Hongbing

2015-08-15

An amperometric glucose biosensor based on direct electron transfer of glucose oxidase (GOD) self-assembled on the surface of partially unzipped carbon nanotubes (PUCNTs) modified glassy carbon electrode (GCE) has been successfully fabricated. PUCNTs were synthesized via a facile chemical oxidative etching CNTs and used as a novel immobilization matrix for GOD. The cyclic voltammetric result of the PUCNT/GOD/GCE showed a pair of well-defined and quasi-reversible redox peaks with a formal potential of -0.470V and a peak to peak separation of 37mV, revealing that the fast direct electron transfer between GOD and the electrode has been achieved. It is notable that the glucose determination has been achieved in mediator-free condition. The developed biosensor displayed satisfactory analytical performance toward glucose including high sensitivity (19.50μA mM(-1)cm(-2)), low apparent Michaelis-Menten (5.09mM), a wide linear range of 0-17mM, and also preventing the interference from ascorbic acid, uric acid and dopamine usually coexisting with glucose in human blood. In addition, the biosensor acquired excellent storage stabilities. This facile, fast, environment-friendly and economical preparation strategy of PUCNT-GOD may provide a new platform for the fabrication of biocompatible glucose biosensors and other types of biosensors. Copyright © 2015 Elsevier B.V. All rights reserved.

Electrochemical affinity biosensors for detection of mycotoxins: A review.

PubMed

Vidal, Juan C; Bonel, Laura; Ezquerra, Alba; Hernández, Susana; Bertolín, Juan R; Cubel, Carlota; Castillo, Juan R

2013-11-15

This review discusses the current state of electrochemical biosensors in the determination of mycotoxins in foods. Mycotoxins are highly toxic secondary metabolites produced by molds. The acute toxicity of these results in serious human and animal health problems, although it has been only since early 1960s when the first studied aflatoxins were found to be carcinogenic. Mycotoxins affect a broad range of agricultural products, most important cereals and cereal-based foods. A majority of countries, mentioning especially the European Union, have established preventive programs to control contamination and strict laws of the permitted levels in foods. Official methods of analysis of mycotoxins normally requires sophisticated instrumentation, e.g. liquid chromatography with fluorescence or mass detectors, combined with extraction procedures for sample preparation. For about sixteen years, the use of simpler and faster analytical procedures based on affinity biosensors has emerged in scientific literature as a very promising alternative, particularly electrochemical (i.e., amperometric, impedance, potentiometric or conductimetric) affinity biosensors due to their simplicity and sensitivity. Typically, electrochemical biosensors for mycotoxins use specific antibodies or aptamers as affinity ligands, although recombinant antibodies, artificial receptors and molecular imprinted polymers show potential utility. This article deals with recent advances in electrochemical affinity biosensors for mycotoxins and covers complete literature from the first reports about sixteen years ago. Copyright © 2013 Elsevier B.V. All rights reserved.

A novel nano-photonics biosensor concept for rapid molecular diagnostics