Download or read book Long wavelength Fluorescence Resonance Energy Transfer FRET based Biosensor for Glucose Sensing Development and Interference Testing written by Mamdouh Salem Aloraefy and published by . This book was released on 2014 with total page 148 pages. Available in PDF, EPUB and Kindle. Book excerpt: Rapid, accurate, and non- or minimally-invasive sensors for glucose measurement have the potential to enhance diabetes control. Recent studies have indicated that implantable optical, affinity biosensors based on Förster Resonance Energy Transfer (FRET) can provide high sensitivity in quantifying glucose concentrations, in continuous and non-destructive fashion. However, there are gaps in the research related to best practices for characterizing the device and factors that alter its performance. First, a standard set of in vitro approaches for evaluating FRET glucose biosensor response has not been established. Second, the potential for chemical interference due to sugars and medications is not well established and standardized methods for quantifying such effects have not been developed. Third, information on the influence of tissue optics on signal detection in these devices is lacking. The general goals of this research project were two-fold: to elucidate the performance and working mechanism of FRET glucose biosensors and identify best practices for assessing performance. Towards these goals, a battery of performance test methods was developed, including spectral response, linearity, sensitivity, limit of detection, kinetic response, reversibility, stability, precision, and accuracy, including error grid analysis. A FRET glucose biosensor was then fabricated and the test methods implemented to fully characterize its response in vitro. Biochemical and optical interference were assessed through a bench-top fluorescence spectroscopy system and phantom measurements, respectively. The biosensor demonstrated a glucose response change of 45%, bias of less than 11%, and a limit of detection of 25 mg/dL. Mannose, maltose, fructose, lactose, and sucrose showed positive results for interference, with concentration estimates over-predicted by up to 64% due to 50 mg/dL concentrations of other sugars. The phantom measurements suggested that non-specific interactions between light and tissue-like samples tended to affect the overall detected signal, with minimal variations in spectral distribution. This was likely due to the choice of fluorophores with long-visible-wavelength emission peaks, where absorption due to hemoglobin is relatively small. The overall results provide evidence of the strengths and weaknesses of the performance of FRET glucose biosensor as well as insight into best practices for thorough objective, quantitative test methods for response evaluation of optical glucose biosensors and factors that influence performance.
Download or read book Fluorescence Based Biosensors written by and published by Academic Press. This book was released on 2012-12-31 with total page 449 pages. Available in PDF, EPUB and Kindle. Book excerpt: One of the major challenges of modern biology and medicine consists in finding means to visualize biomolecules in their natural environment with the greatest level of accuracy, so as to gain insight into their properties and behaviour in a physiological and pathological setting. This has been achieved thanks to the design of novel imaging agents, in particular to fluorescent biosensors. Fluorescence Biosensors comprise a large set of tools which are useful for fundamental purposes as well as for applications in biomedicine, drug discovery and biotechnology. These tools have been designed and engineered thanks to the combined efforts of chemists and biologists over the last decade, and developed hand in hand together with imaging technologies. This volume will convey the many exciting developments the field of fluorescent biosensors and reporters has witnessed over the recent years, from concepts to applications, including chapters on the chemistry of fluorescent probes, on technologies for monitoring protein/protein interactions and technologies for imaging biosensors in cultured cells and in vivo. Other chapters are devoted to specific examples of genetically-encoded reporters, or to protein and peptide biosensors, together with examples illustrating their application to cellular and in vivo imaging, biomedical applications, drug discovery and high throughput screening. - Contributions from leading authorities - Informs and updates on all the latest developments in the field
Download or read book Fluorescence Sensors and Biosensors written by Richard B. Thompson and published by CRC Press. This book was released on 2005-12-16 with total page 416 pages. Available in PDF, EPUB and Kindle. Book excerpt: Fluorescence-based sensing is a significant technique used in prominent fields such as fluorescence-activated cell sorting, DNA sequencing, high-throughput screening, and clinical diagnostics. Fluorescence Sensors and Biosensors emphasizes the most recent developments and emerging technologies with the broadest impacts. The text begins wi
Download or read book Glucose Sensing written by Chris D. Geddes and published by Springer Science & Business Media. This book was released on 2007-12-29 with total page 460 pages. Available in PDF, EPUB and Kindle. Book excerpt: An essential reference for any laboratory working in the analytical fluorescence glucose sensing field. The increasing importance of these techniques is typified in one emerging area by developing non-invasive and continuous approaches for physiological glucose monitoring. This volume incorporates analytical fluorescence-based glucose sensing reviews, specialized enough to be attractive to professional researchers, yet appealing to a wider audience of scientists in related disciplines of fluorescence.
Download or read book Towards a Fluorescence Resonance Energy Transfer FRET Strategy for Development of a Biosensor for Detection of Nucleic Acid Hybridization on Fused Silica Surfaces microform written by Melissa Massey and published by Library and Archives Canada = Bibliothèque et Archives Canada. This book was released on 2005 with total page 368 pages. Available in PDF, EPUB and Kindle. Book excerpt: Thiazole orange (TO) with different tethers were synthesized to be attached to oligonucleotides. The FRET of TO in solution with double-stranded DNA (dsDNA) was investigated with BlackHole (BHQ1) or ((4-dimethylamino)phenyl)azo)benzoic acid (DABCYL) quenchers which decreased the fluorescence 2.9 +/- 7% and 2.5 +/- 10% times, respectively. A quenching mechanism could therefore be designed to transduce hybridization. The FRET of N, N, N, N-tetramethylcarboxyrhodamine (TAMRA) and IowaBlackRQ RTM (IABLK) linked to complementary oligonucleotides immobilized on glass substrates was investigated; IABLK quenched TAMRA fluorescence. However, surface bound dsDNA caused some self-quenching of TAMRA. Solution FRET using TAMRA/IABLK at 24.5°C and 60°C with complementary and mismatched DNA was measured to investigate potential for mismatch detection. The probe sequence was based on the determinate for Spinal Muscular Atrophy (SMA). Signal intensity differed between complementary and the mismatch samples at 60°C, indicating mismatch detection potential. Results suggest the possibility of designing tethered fluorophore-quencher pairs for transduction of hybridization for development of optical nucleic acid biosensors for SMA screening.
Download or read book Fluorescence Resonance Energy Transfer FRET Based Sensors for Bioanalysis written by Gabriela Blagoi and published by . This book was released on 2004 with total page pages. Available in PDF, EPUB and Kindle. Book excerpt: The objective of my PhD study was to develop and characterize new methods and sensors based on fluorescence resonance energy transfer (FRET) for bioanalysis. Chapter 3 describes the use of FRET between donor fluorophores and acceptor labeled murine macrophage cells. FRET microscopy was used to determine whether the donor molecules truly permeate through the cell membrane or only adsorb to the cell surface. This method was found to be partially successful since the donor red tail fluorescence overlapped with the sensitized acceptor fluorescence and led to false reading of FRET. We found that is easier to monitor delivery of acceptor molecules into donor-labeled cells. Using donor labeled cells it was possible to determine whether the acceptor molecules were actually delivered into cells. However, a relatively high acceptor concentration in the hundreds of micromolar level was needed to obtain measurable FRET signals in the 3-D cellular system. The results underscored the need to reduce the dimensionality of FRET systems in order to increase the FRET efficiency between donor and acceptor molecules. Chapter 4 describes the development of FRET sensing lipobeads labeled with donors and their use to evaluate the interactions of acceptor molecules with the phospholipid membrane of FRET sensing lipobeads. The change in the dimensionality of the system in which FRET occurs, improved the sensitivity of our measurements by 3-folds compared to FRET measurements in solution. We concluded that a molecular recognition component had to be added to the sensing particles to further increase their selectivity and sensitivity. Chapter 5 describes the development of FRET trap sensing beads and their use for screening nonfluorescent carbohydrates and glycoproteins. The FRET sensing technique was based on binding between dextran molecules labeled with Texas Red (Dextran-TR) and polystyrene microparticles labeled with Fluorescein tagged Concanavalin A (FITC-ConA). It was found that carbohydrates and glycoproteins inhibit the binding between dextran-TR and FITC-ConA labeled particles. The inhibition effect was concentration dependent thus enabled screening carbohydrates and glycoproteins based on their inhibition potency. The dissertation critically evaluates the performance of FRET microscopy and FRET based sensors in delivery and screening applications.
Download or read book Application and Modeling of Frequency domain Lifetime Spectroscopy for Microsphere based Optical Glucose Sensors written by Feng Liang and published by . This book was released on 2010 with total page pages. Available in PDF, EPUB and Kindle. Book excerpt: A new glucose affinity sensor based on a homogeneous fluorescence resonance energy transfer (FRET) assay system was developed to monitor the competitive binding between concanavalin A (ConA) and dextran. The FRET quenching kinetics of the donor were analyzed from frequency-domain (FD) measurements as functions of both glucose and acceptor-protein concentrations using a Förster-type decay kinetics model. The results showed that the FD measurements and donor decay kinetics can indicate quantitative changes in the presence of glucose at concentrations ranging from 0 to 224 mg/dL. The second set of experiments proved the feasibility of performing analyte sensing with FD lifetime spectroscopy using microsphere-based sensors in multiple scattering solutions. A well characterized pH-sensitive fluorophore was entrapped in poly(ethylene glycol) microspheres. The particles were then immersed in a buffered polystyrene solution of various pH. Measurements of phase shift and modulation of the generated and multiply scattered fluorescent light were acquired as the modulation frequency of the incident excitation light varied from 10 to 120 MHz. After the measured data were analyzed with the coupled diffusion equations, the obtained lifetimes from the scattering measurements matched values from non-scattering measurements. Lastly, a new two-speed Monte Carlo (MC) simulation was developed to predict light propagation through the sensors and thus was used to evaluate these sensors and to design these sensors for implantation. The model used random packing structure and considered geometric optics and two light propagation speeds. Experimental measurements of phase-shift and modulation of excitation light were made on a cubic phantom with non-fluorescent resin microspheres of 74 ℗æm diameter, and compared to those computed from the MC simulation. The results showed that the mean phase shift (PS) deviation was 0.736° and the mean amplitude deviation was 42%. Quantitative changes in detected fluorescence phase-shift and modulation were investigated for microsphere diameter, volume fraction, refractive index, and fluorophore lifetime. We also found that even though the sensitivity of PS change in the presence of scattering was the same as the value without scattering, the values of PS were magnified due to the scattering effects.
Download or read book Development of a Fluorescence Resonance Energy Transfer Optical Nanoscale Biosensor Based on a Liquid core Waveguide Platform written by R. Cody Stringer and published by . This book was released on 2007 with total page pages. Available in PDF, EPUB and Kindle. Book excerpt: In order to produce a more versatile, adaptable, and effective method for detection of biological analytes, a self-contained and robust fluorescent optical biosensor architecture utilizing fluorescence resonance energy transfer (FRET) is proposed. This biosensor architecture is then applied to a highly adept liquid-core waveguide platform. FRET is a distance-dependent signal transduction method that occurs between two fluorescent molecules, termed the donor and acceptor. When the donor and acceptor are brought within close proximity, a quantifiable nonradiative energy exchange takes place. In order to launch FRET, a donor-labeled Protein A molecule is bound to an acceptor-labeled capture antibody. When exposed to antigen, the antibody-antigen binding event initiates a conformational change within the structure of the antibody, and thereby induces a measurable change in energy transfer from the donor to the acceptor by altering the distance between the FRET pair. Additionally, effects of quantum dots and gold nanoparticles utilized within the FRET system are studied. The resulting system is then optimized and tested in a liquid-core waveguide platform that is able to retrieve sensitive and accurate measurements. In the current study, the biosensor was used to detect Porcine Reproductive and Respiratory Syndrome virus and human cardiac Troponin I, showing ample sensitivity and a high degree of specificity, as well as rapid response.
Download or read book Designing Microwave Sensors for Glucose Concentration Detection in Aqueous and Biological Solutions written by Carlos G. Juan and published by Springer Nature. This book was released on 2021-06-14 with total page 256 pages. Available in PDF, EPUB and Kindle. Book excerpt: This book presents a comprehensive study covering the design and application of microwave sensors for glucose concentration detection, with a special focus on glucose concentration tracking in watery and biological solutions. This book is based on the idea that changes in the glucose concentration provoke variations in the dielectric permittivity of the medium. Sensors whose electrical response is sensitive to the dielectric permittivity of the surrounding media should be able to perform as glucose concentration trackers. At first, this book offers an in-depth study of the dielectric permittivity of water–glucose solutions at concentrations relevant for diabetes purposes; in turn, it presents guidelines for designing suitable microwave resonators, which are then tested in both water–glucose solutions and multi-component human blood plasma solutions for their detection ability and sensitivities. Finally, a portable version is developed and tested on a large number of individuals in a real clinical scenario. All in all, the book reports on a comprehensive study on glucose monitoring devices based on microwave sensors. It covers in depth the theoretical background, provides extensive design guidelines to maximize sensitivity, and validates a portable device for applications in clinical settings.
Download or read book Resonant Energy Transfer Based Biosensor for Detection of Multivalent Proteins written by and published by . This book was released on 2001 with total page 9 pages. Available in PDF, EPUB and Kindle. Book excerpt: We have developed a new fluorescence-based biosensor for sensitive detection of species involved in a multivslent interaction. The biosensor system utilizes specific interactions between proteins and cell surface receptors, which trigger a receptor aggregation process. Distance-dependent fluorescence self-quenching and resonant energy transfer mechanisms were coupled with a multivalent interaction to probe the receptor aggregation process, providing a sensitive and specific signal transduction method for such a binding event. The fluorescence change induced by the aggregation process can be monitored by different instrument platforms, e.g. fluorimetry and flow cytometry. In this article, a sensitive detection of pentavalent cholera toxin which recognizes ganglioside GM1 has been demonstrated through the resonant energy transfer scheme, which can achieve a double color change simultaneously. A detection sensitivity as high as 10 pM has been achieved within a few minutes (c.a. 5 minutes). The simultaneous double color change (an increase of acceptor fluorescence and a decrease of donor fluorescence intensity) of two similar fluorescent probes provides particularly high detection reliability owing to the fact that they act as each other's internal reference. Any external perturbation such as environmental temperature change causes no significant change in signal generation. Besides the application for biological sensing, the method also provides a useful tool for investigation of kinetics and thermodynamics of a multivalent interaction. Keywords: Biosensor, Fluorescence resonant energy transfer, Multivalent interaction, Cholera Toxin, Ganglioside GM1, Signal Transduction.
Download or read book Handbook of Optical Sensing of Glucose in Biological Fluids and Tissues written by Valery V. Tuchin and published by CRC Press. This book was released on 2008-12-22 with total page 744 pages. Available in PDF, EPUB and Kindle. Book excerpt: Although noninvasive, continuous monitoring of glucose concentration in blood and tissues is one of the most challenging areas in medicine, a wide range of optical techniques has recently been designed to help develop robust noninvasive methods for glucose sensing. For the first time in book form, the Handbook of Optical Sensing of Glucose in Biological Fluids and Tissues analyzes trends in noninvasive optical glucose sensing and discusses its impact on tissue optical properties. This handbook presents methods that improve the accuracy in glucose prediction based on infrared absorption spectroscopy, recent studies on the influence of acute hyperglycemia on cerebral blood flow, and the correlation between diabetes and the thermo-optical response of human skin. It examines skin glucose monitoring by near-infrared spectroscopy (NIR), fluorescence-based glucose biosensors, and a photonic crystal contact lens sensor. The contributors also explore problems of polarimetric glucose sensing in transparent and turbid tissues as well as offer a high-resolution optical technique for noninvasive, continuous, and accurate blood glucose monitoring and glucose diffusion measurement. Written by world-renowned experts in biomedical optics and biophotonics, this book gives a complete, state-of-the-art treatise on the design and applications of noninvasive optical methods and instruments for glucose sensing.
Download or read book Toward Multiplexed Nucleic Acid Assays and Biosensors Using Immobilized Quantum Dots as Donors in Fluorescence Resonance Energy Transfer FRET written by Walter Russell Algar and published by . This book was released on 2010 with total page pages. Available in PDF, EPUB and Kindle. Book excerpt:
Download or read book Development of New Fluorescent Protein Biosensors written by Ahmed Saied F. Belal and published by . This book was released on 2014 with total page 124 pages. Available in PDF, EPUB and Kindle. Book excerpt: The discovery of green fluorescent protein (GFP) from the Aequorea victoria Jellyfish followed by the extensive efforts of protein engineers to produce other fluorescent proteins (FPs) spanning the visible color spectrum made fluorescent proteins indispensable biochemical tools in the scientific community. Experimental biologists have utilized FPs as genetically encoded markers for the imaging of subcellular structures and protein dynamics in live cells. Another important application of FPs is their design for use in biosensors for either enzymes or small biological molecules of interest. The work described in this thesis is an attempt to portray different experimental designs of FP based biosensors with the final objective of either modifying previously reported or introducing novel biosensors. We addressed the FP biosensors based on the principle of intramolecular Förster resonance energy transfer (FRET) in two projects. In the first project we demonstrated a modification of a methodology of development and optimization of FRET-based biosensor for a post translational modification. The end result of this project has led to improving a previously reported protein kinase B (PkB) biosensor and the discovery of a new cyclin B1- cyclin dependent kinase 1 (Cyclin B1-CDK1) biosensor of a higher dynamic range than previously published one. In the second project our efforts were directed to develop a matrix metallo proteinase 2 (MMP2) FRET based biosensor, with the ultimate goal of using this biosensor in live cell imaging of cardiomyocytes to explore postulated MMP2 intracellular role in ischemia-reperfusion injury. We designed protein constructs based on both previously reported MMP2 substrate sequences and potential cardiac protein target sequences. After testing and characterizing the designed constructs, the expression of the best candidate in neonatal cardiomyocytes cell lines is undertaken by our collaborator Professor Richard Schulz, Department of Pharmacology, University of Alberta. In the third and final project we utilized single FP-based biosensor concept in trying to develop Hydrogen peroxide biosensors of different hues. Our efforts were fruitful in discovering two green biosensors, one is with direct and the other is with inverse response to hydrogen peroxide.
Download or read book Quantum Dot fluorescent Protein Pairs as Fluorescence Resonance Energy Transfer Pairs written by Allison Marie Dennis and published by . This book was released on 2009 with total page pages. Available in PDF, EPUB and Kindle. Book excerpt: Fluorescence resonance energy transfer (FRET)-based biosensors have been designed to fluorometrically detect everything from proteolytic activity to receptor-ligand interactions and structural changes in proteins. While a wide variety of fluorophores have demonstrated effectiveness in FRET probes, several potential sensor components are particularly notable. Semiconductor quantum dots (QDs) are attractive FRET donors because they are rather bright, exhibit high quantum yields, and their nanoparticulate structure enables the attachment of multiple acceptor molecules. Fluorescent proteins (FPs) are also of particular interest for fluorescent biosensors because design elements necessary for signal transduction, probe assembly, and device delivery and localization for intracellular applications can all be genetically incorporated into the FP polypeptide. : The studies described in this thesis elucidate the important parameters for concerted QD-FP FRET probe design. Experimental results clarify issues of FRET pair selection, probe assembly, and donor-acceptor distance for the multivalent systems. Various analysis approaches are compared and guidelines asserted based on the results. To demonstrate the effectiveness of the QD-FP FRET probe platform, a ratiometric pH sensor is presented. The sensor, which uses the intrinsic pH-sensitivity of the FP mOrange to modulate the FP/QD emission ratio, exhibits a 20-fold change in its ratiometric measurement over a physiologically interesting pH range, making it a prime candidate for intracellular imaging applications.
Download or read book Optical Glucose Nanobiosensor Encapsulated in Erythrocytes written by Majed El-Dweik and published by . This book was released on 2007 with total page pages. Available in PDF, EPUB and Kindle. Book excerpt: An implantable glucose biosensor encapsulated in erythrocytes, Red Blood Cells (RBC), will become a method for continuously measuring blood glucose concentration in diabetics. In 2005, the American Diabetes Association (ADA) reported that 20.8 million people have diabetes, making it the fifth leading cause of death by disease in the USA. This paper focuses on the preparation phase of the glucose sensor. Glucose Binding Protein (GBP) from E. coli was labeled with two fluorophores, Alexa Fluor 680 (AF680), and Alexa Fluor 750 (AF750). This sensor based on Fluorescence Resonance Energy Transfer (FRET). FRET is a distance sensitive technique between the above fluorophores. The initial energy transfer between AF680 and AF750 labeled on the GBP before glucose additions was determined. After glucose additions, the labeled GBP went through conformational change which caused distance between the labeled sites. This change in distance caused a change in the energy transfer. The labeled GBP became the glucose nanobiosensor. The labeled GBP nanobiosensors were encapsulated in erythrocytes, red blood cells (RBCs), by using the Hypo-Osmotic dialysis technique. The encapsulated RBCs responded well to different glucose concentrations ranging form 0-33.16mM. This range covers the normal blood glucose concentration, 4 - 9mM.
Download or read book In Vivo Glucose Sensing written by David D. Cunningham and published by John Wiley & Sons. This book was released on 2009-11-19 with total page 466 pages. Available in PDF, EPUB and Kindle. Book excerpt: In Vivo Glucose Sensing is a key reference for scientists and engineers working on the development of glucose sensing technologies for the management of diabetes and other medical conditions. It discusses the analytical chemistry behind the strategies currently used for measuring glucose in vivo. It focuses on analyzing samples in the real world and discusses the biological complexities that make glucose sensing difficult. Covering current implantable devices, next-generation implantable sensing methods, and non-invasive methods for measuring glucose, this book concludes with an overview of possible applications other than diabetes.
Download or read book Biosensor Technology written by Buck and published by Routledge. This book was released on 2017-11-22 with total page 408 pages. Available in PDF, EPUB and Kindle. Book excerpt: This authoritative reference covers recent advances in the field, stressing an interdisciplinaryapproach to the development and use of biosensor technology in physics,engineering, analytical chemistry, and biochemistry (including immunochemistry).about the editors ...RICHARD P. BucK is a Professor in the Chemistry Department, University of Northcarolina, Chapel Hill. Professor Buck serves on the editorial boards of severaljournals including Analytical Instrumentation: Applications and Designs for Chemica~Biomedica~ and Environmental Science (Marcel Dekker, Inc.). He is a member of theAmerican Chemical Society, Electrochemical Society, and International Society ofElectrochemistry. He received the B.S. (1950) and M.S. (1951) degrees from thecalifornia Institute of Tochnology, Pasadena, and Ph.D. degree (1954) from theMassachusetts Institute of Tochnology, cambridge.W1WAM E. HATFIELD is Mary Ann Smith Professor and Vice Chairman of Chemistry,and Acting Chairman of the Curriculum in Applied Sciences, University of Northcarolina, Chapel Hill. He is the author or coauthor of over 300 publications, andcoeditor, with John H. Miller, Jr., of High-Temperature Superconducting Materials:Preparations, Properties, and Processing (Marcel Dekker, Inc.). He is a member of theAmerican Chemical Society, American Association for the Advancement of Science,and Materials Research Society. He received the B.S. (1958) and M.S. (1959) degreesfrom Marshall University, Huntington, West Virginia, Ph.D. degree (1962) from theUniversity of Arizona, Tucson, and completed postdoctoral research at the Universityof Illinois, Urbana.M1KTHA UMANA is an independent consultant to Glaxo Inc. and Research 'IriangleInstitute, Research 'Iriangle Park, and Duke University Engineering Research Center,Durham, North carolina. The coauthor of numerous scientific journal articles, herresearch interests include surface chemistry, electrochemistry, and biosensors. Shereceived the B.Sc. degree (1969) from the University of Chile, Santiago, and Ph.D.degree (1972) from the University of London, England.EDMOND E BowoEN is an Associate Professor in the Department of Chemistry andin the Biotechnology Program, North carolina State University, Raleigh. Thecoauthor of numerous journal articles, his research interests include bioelectrochemistry,biological electron transfer, biosensors, and surface chemistry. He receivedthe B.S. degree (1970) from Syracuse University, Syracuse, New York, and Ph.D.degree (1982) from Virginia Commonwealth University, Richmond.
