Towards A Microfluidics Based Nucleic Acid Biosensor Using Immobilized Quantum Dot Dna Conjugates For Fret Detection Of Target Oligonucleotides

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Towards a Microfluidics-based Nucleic Acid Biosensor Using Immobilized Quantum Dot - DNA Conjugates for FRET Detection of Target Oligonucleotides

Author : Jonathan Cauchi
Publisher :
ISBN 13 :
Total Pages : 0 pages
Book Rating : 4.:/5 (133 download)

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Book Synopsis Towards a Microfluidics-based Nucleic Acid Biosensor Using Immobilized Quantum Dot - DNA Conjugates for FRET Detection of Target Oligonucleotides by : Jonathan Cauchi

Download or read book Towards a Microfluidics-based Nucleic Acid Biosensor Using Immobilized Quantum Dot - DNA Conjugates for FRET Detection of Target Oligonucleotides written by Jonathan Cauchi and published by . This book was released on 2009 with total page 0 pages. Available in PDF, EPUB and Kindle. Book excerpt: The potential for an electrokinetically driven, FRET-based microfluidic biosensor for SNP discrimination has been explored. The method functionalized the glass wall of a PDMS/glass microfluidics channel with multidentate thiol ligands to noncovalently immobilize MPA capped CdSe/ZnS QDs. Single stranded probe DNA could then be immobilized to QDs and target material could be delivered electrokinetically to the sensing surface. SNP discrimination could then occur by manipulation of shear, electrical and thermal effects derived from the applied voltage. The stability of immobilized QDs was investigated by EOF experiments that applied 500 V and 100 V voltages for 10 minutes to initiate electrokintetic flow. Fluorescence intensity measurements showed nearly complete removal of QDs from slides when compared with controls at both voltages. Pressure driven flow experiments demonstrated reduced dissociation of immobilized of QDs compared to channels exposed to EOF. A covalent approach is likely necessary to ensure stability of immobilized QDs during EOF.

Toward Multiplexed Nucleic Acid Assays and Biosensors Using Immobilized Quantum Dots as Donors in Fluorescence Resonance Energy Transfer (FRET).

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Author : Walter Russell Algar
Publisher :
ISBN 13 :
Total Pages : 696 pages
Book Rating : 4.:/5 (747 download)

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Book Synopsis Toward Multiplexed Nucleic Acid Assays and Biosensors Using Immobilized Quantum Dots as Donors in Fluorescence Resonance Energy Transfer (FRET). by : Walter Russell Algar

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 696 pages. Available in PDF, EPUB and Kindle. Book excerpt: Research toward a multiplexed nucleic acid biosensor that uses quantum dots (QDs) as donors in a fluorescence resonance energy transfer (FRET) assay is described. Optical fibers were modified with mixed films composed of different colours of QDs and different oligonucleotide probes that served as scaffolds for the hybridization of the corresponding target nucleic acid sequences. Fluorescent dyes that were suitable as acceptors for each QD donor were associated with hybridization and provided an analytical signal through FRET-sensitized emission. Different detection channels were achieved through the combination of different donors and acceptors: green emitting QDs with Cyanine 3 or Rhodamine Red-X; and red emitting QDs with Alexa Fluor 647. A detection channel that used the direct excitation of Pacific Blue complemented the FRET pairs. One-plex, two-plex, three-plex and four-plex hybridization assays were demonstrated. A sandwich assay format was adopted to avoid target labeling. Detection limits were 1-10 nM (1-12 pmol) and analysis times were 1-4 h. Single nucleotide polymorphisms were discriminated in multiplexed assays, and the potential for reusability was also demonstrated. Non-selective interactions between QDs and oligonucleotides were characterized, and routes toward the optimization of the QD-FRET hybridization assays were identified. A basic model for multiple FRET pathways in a mixed film was also developed. In addition to the advantages of solid-phase assays, the combination of QDs and FRET was advantageous because it permitted multiplexed detection using a single excitation source and a single substrate, in the ensemble, and via ratiometric signals. Spatial registration or sorting methods, imaging or spatial scanning, and single molecule spectroscopy were not required. The research in this thesis is expected to enable new chip-based biosensors in the future, and is an original contribution to both bioanalytical spectroscopy and the bioanalytical applications of nanomaterials.

Quantum Dots for DNA Biosensing

Author : Jun-Jie Zhu
Publisher : Springer Science & Business Media
ISBN 13 : 3642449107
Total Pages : 96 pages
Book Rating : 4.6/5 (424 download)

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Book Synopsis Quantum Dots for DNA Biosensing by : Jun-Jie Zhu

Download or read book Quantum Dots for DNA Biosensing written by Jun-Jie Zhu and published by Springer Science & Business Media. This book was released on 2013-12-04 with total page 96 pages. Available in PDF, EPUB and Kindle. Book excerpt: This book provides a broad introduction to all major aspects of quantum dot properties including fluorescence, electrochemical, photochemical and electroluminescence. Such properties have been produced for applications in biosensing, cell tracking, in vivo animal imaging and so on. It focuses on their special applications in DNA biosensing and provides readers with detailed information on the preparation and functionalization of quantum dots and the fabrication of DNA biosensors, using examples to show how these properties can be used in DNA biosensor design and the advantages of quantum dots in DNA biosensing. Further new emerging quantum dots such as metal nanoclusters and graphene dots and their applications in DNA biosensing have also been included.

Towards The Development Of A Quantum Dot Based Bioprobe For Intracellular Investigations of Nucleic Acid Hybridization Events Using Fluorescence Resonance Energy Transfer

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Author : Lori Lok-Yin Chong
Publisher :
ISBN 13 : 9780494764756
Total Pages : 268 pages
Book Rating : 4.7/5 (647 download)

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Book Synopsis Towards The Development Of A Quantum Dot Based Bioprobe For Intracellular Investigations of Nucleic Acid Hybridization Events Using Fluorescence Resonance Energy Transfer by : Lori Lok-Yin Chong

Download or read book Towards The Development Of A Quantum Dot Based Bioprobe For Intracellular Investigations of Nucleic Acid Hybridization Events Using Fluorescence Resonance Energy Transfer written by Lori Lok-Yin Chong and published by . This book was released on 2011 with total page 268 pages. Available in PDF, EPUB and Kindle. Book excerpt: The unique spectroscopic properties of quantum dots (QDs) are of interest for application in intracellular studies of gene expression. QDs derivatized with single-stranded probe oligonucleotides were used to detect complementary target sequences via hybridization and fluorescence resonance energy transfer (FRET). As nucleic acid targets are not labeled within cells, a displacement assay for nucleic acid detection featuring QDs as FRET donors was developed. QDs conjugated with oligonucleotide probes and then pre-hybridized with labeled target yielded efficient FRET in vitro. Studies in vitro confirmed that displacement kinetics of pre-hybridized target was a function of the stability of the initial hybridized complex. Displacement was observed as reduction in FRET intensity coupled with regeneration of QD fluorescence. By engineering the sequence of the labeled target, faster displacement was possible. The QD-probe+target system was successfully delivered into cells via transfection. Although QDs with their cargo remained sequestered in endosomal vesicles, fluorescent properties were retained.

DNA Hybridization on Walls of Electrokinetically Controlled Microfluidic Channels

Author : Lu Chen
Publisher :
ISBN 13 :
Total Pages : pages
Book Rating : 4.:/5 (13 download)

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Book Synopsis DNA Hybridization on Walls of Electrokinetically Controlled Microfluidic Channels by : Lu Chen

Download or read book DNA Hybridization on Walls of Electrokinetically Controlled Microfluidic Channels written by Lu Chen and published by . This book was released on 2010 with total page pages. Available in PDF, EPUB and Kindle. Book excerpt:

Microfluidic Biosensors

Author : Wing Cheung Mak
Publisher : Elsevier
ISBN 13 : 012823847X
Total Pages : 370 pages
Book Rating : 4.1/5 (282 download)

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Book Synopsis Microfluidic Biosensors by : Wing Cheung Mak

Download or read book Microfluidic Biosensors written by Wing Cheung Mak and published by Elsevier. This book was released on 2022-11-05 with total page 370 pages. Available in PDF, EPUB and Kindle. Book excerpt: Microfluidic Biosensors provides a comprehensive overview covering the most recent emerging technologies on the design, fabrication, and integration of microfluidics with transducers. These form various integrated microfluidic biosensors with device configurations ranging from 2D to 4D levels. Coverage also includes advanced printed microfluidic biosensors, flexible microfluidics for wearable biosensors, autonomous lab-on-a-chip biosensors, CMOS-base microanalysis systems, and microfluidic devices for mobile phone biosensing. The editors and contributors of this book represent both academia and industry, come from a varied range of backgrounds, and offer a global perspective. This book discusses the design and principle of microfluidic systems and uses them for biosensing applications. The microfluidic fabrication technologies covered in this book provide an up-to-date view, allowing the community to think of new ways to overcome challenges faced in this field. The focus is on existing and emerging technologies not currently being analyzed extensively elsewhere, providing a unique perspective and much-needed content. The editors have crafted this book to be accessible to all levels of academics from graduate students, researchers, and professors working in the fields of biosensors, microfluidics design, material science, analytical chemistry, biomedical devices, and biomedical engineering. It can also be useful for industry professionals working for microfluidic device manufacturers, or in the industry of biosensors and biomedical devices. Presents an in-depth overview of microfluidic biosensors and associated emerging technologies such as printed microfluidics and novel transducers Addresses a range of microfluidic biosensors with device configurations ranging from 2D to 4D levels Includes the commercialization aspects of microfluidic biosensors that provide insights for scientists and engineers in research and development

Microfluidics and Nanotechnology

Author : Eric Lagally
Publisher : CRC Press
ISBN 13 : 1351831488
Total Pages : 294 pages
Book Rating : 4.3/5 (518 download)

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Book Synopsis Microfluidics and Nanotechnology by : Eric Lagally

Download or read book Microfluidics and Nanotechnology written by Eric Lagally and published by CRC Press. This book was released on 2017-12-19 with total page 294 pages. Available in PDF, EPUB and Kindle. Book excerpt: An increasing number of technologies are being used to detect minute quantities of biomolecules and cells. However, it can be difficult to determine which technologies show the most promise for high-sensitivity and low-limit detection in different applications. Microfluidics and Nanotechnology: Biosensing to the Single Molecule Limit details proven approaches for the detection of single cells and even single molecules—approaches employed by the world’s foremost microfluidics and nanotechnology laboratories. While similar books concentrate only on microfluidics or nanotechnology, this book focuses on the combination of soft materials (elastomers and other polymers) with hard materials (semiconductors, metals, and glass) to form integrated detection systems for biological and chemical targets. It explores physical and chemical—as well as contact and noncontact—detection methods, using case studies to demonstrate system capabilities. Presenting a snapshot of the current state of the art, the text: Explains the theory behind different detection techniques, from mechanical resonators for detecting cell density to fiber-optic methods for detecting DNA hybridization, and beyond Examines microfluidic advances, including droplet microfluidics, digital microfluidics for manipulating droplets on the microscale, and more Highlights an array of technologies to allow for a comparison of the fundamental advantages and challenges of each, as well as an appreciation of the power of leveraging scalability and integration to achieve sensitivity at low cost Microfluidics and Nanotechnology: Biosensing to the Single Molecule Limit not only serves as a quick reference for the latest achievements in biochemical detection at the single-cell and single-molecule levels, but also provides researchers with inspiration for further innovation and expansion of the field.

Towards a Fluorescence Resonance Energy Transfer (FRET) Strategy for Development of a Biosensor for Detection of Nucleic Acid Hybridization on Fused Silica Surfaces [microform]

Download Towards a Fluorescence Resonance Energy Transfer (FRET) Strategy for Development of a Biosensor for Detection of Nucleic Acid Hybridization on Fused Silica Surfaces [microform] PDF Online Free

Author : Melissa Massey
Publisher : Library and Archives Canada = Bibliothèque et Archives Canada
ISBN 13 : 9780494021569
Total Pages : 368 pages
Book Rating : 4.0/5 (215 download)

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Book Synopsis Towards a Fluorescence Resonance Energy Transfer (FRET) Strategy for Development of a Biosensor for Detection of Nucleic Acid Hybridization on Fused Silica Surfaces [microform] by : Melissa Massey

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.

Towards a Microfluidic Nucleic Acid Biosensor [microform] : Use of Photochemistry for Immobilization of a Density Gradient of Probes

Author : Sara A. (Sara Anne) Dicks
Publisher : Library and Archives Canada = Bibliothèque et Archives Canada
ISBN 13 : 9780612952249
Total Pages : 300 pages
Book Rating : 4.9/5 (522 download)

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Book Synopsis Towards a Microfluidic Nucleic Acid Biosensor [microform] : Use of Photochemistry for Immobilization of a Density Gradient of Probes by : Sara A. (Sara Anne) Dicks

Download or read book Towards a Microfluidic Nucleic Acid Biosensor [microform] : Use of Photochemistry for Immobilization of a Density Gradient of Probes written by Sara A. (Sara Anne) Dicks and published by Library and Archives Canada = Bibliothèque et Archives Canada. This book was released on 2004 with total page 300 pages. Available in PDF, EPUB and Kindle. Book excerpt: A class of photolabile protecting groups, namely nitrobenzyl derivatives were investigated for the purpose of finding a suitable reactive coupling group to be used in the manufacture of slides for the development of a microfluidic nucleic acid biosensor. The ideal photolabile protecting group would enable quantitative control of the covalent immobilization chemistry used for attachment of nucleic acid probes on the biosensor surface. Nitrobenzyl bromide, a nitrobenzyl-ether olefin siloxane and a nitrobenzyl-ester olefin siloxane were the systems of study. While the three systems all could provide for covalent coupling, none provided sufficient control to permit practical and reproducible applications in the development of the biosensor. With respect to the nitrobenzyl bromide system, the stoichometry could not be controlled and the extreme water sensitivity was a problem. The ether and ester systems required excessive deprotection reaction time, even when exposed to intense UV radiation and did not exhibit sufficient reproducibility.

Development of a Fibre Optic Biosensor for Determination of Interfacial Nucleic Acid Hybrid Formation

Author : Paul Angelo Emilio Piunno
Publisher :
ISBN 13 :
Total Pages : 0 pages
Book Rating : 4.:/5 (133 download)

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Book Synopsis Development of a Fibre Optic Biosensor for Determination of Interfacial Nucleic Acid Hybrid Formation by : Paul Angelo Emilio Piunno

Download or read book Development of a Fibre Optic Biosensor for Determination of Interfacial Nucleic Acid Hybrid Formation written by Paul Angelo Emilio Piunno and published by . This book was released on 1999 with total page 0 pages. Available in PDF, EPUB and Kindle. Book excerpt: This work has demonstrated that single-stranded DNA may be assembled by solid-phase oligonucleotide synthesis onto linker functionalised fused silica optical fibres and can participate in hybrid formation with complementary DNA and RNA. Hybridisation events may be detected through the use of the fluorescent intercalant dye ethidium bromide (EtBr) to provide an optical sensor capable for nucleic acid determination. Four strategies for the functionalisation of substrates with oligonucleotide-hexa(ethylene glycol) (BEG) conjugates were investigated by quantitative AE-HPLC following liberation of the conjugate from the solid support. The method for sensor functionalisation employing pretreatment of the substrate surface with glycidoxypropyltrimethoxysilane followed by extension with dimethoxytrityl protected BEG proved most practical owing to the stability of the immobilisation chemistry and the control provided over strand immobilisation. Optical sensors have been prepared for various target nucleic acids, including sequences known to be selective for the identification of the pathogenic fungi 'Candida albicans', pathogenic forms of the bacterium ' Salmonella', and for genetically modified organisms, such as Round-Up ReadyTm Soy beans. An automated intrinsic mode fibre optic spectrofluorimeter was developed for fluorimetric analysis of interfacial hybrid formation on the optical sensors. Selective, sensitive, rapid, reproducible, and quantitative determinations have been made using the dedicated instrument and optical sensors developed in this work. The sensors have also been observed to be long-lived and rugged. Experiments done to determine the availability of the immobilised strands for participation in complementary binding indicated that the hybridisation efficiency for the immobilised strands was quantitative. Detection of triple-helical nucleic acid formation on the surface of the optical sensors developed in this work has also been demonstrated. A fundamental physical study has also been done that explored the ramifications of the use of immobilised DNA layers as selective reagents. Preliminary work has been done to create a reagentless biosensor. An analogue of EtBr tethered by a 19-atom chain to the termini of immobilised oligonucleotides was prepared to demonstrate reagentless sensor operation with accelerated response times. Further work has focused on the creation of fluorescent dyes that may be tethered to immobilised oligonucleotides functionalised with a polyether tether at the 5'-terminus.

Electrogenerated Chemiluminescence

Author : Allen J. Bard
Publisher : CRC Press
ISBN 13 : 1135535795
Total Pages : 707 pages
Book Rating : 4.1/5 (355 download)

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Book Synopsis Electrogenerated Chemiluminescence by : Allen J. Bard

Download or read book Electrogenerated Chemiluminescence written by Allen J. Bard and published by CRC Press. This book was released on 2004-07-20 with total page 707 pages. Available in PDF, EPUB and Kindle. Book excerpt: The first source on this expanding analytical science, this reference explores advances in the instrumentation, design, and application of techniques with electrogenerated chemiluminescence (ECL), examining the use and impact of ECL-based assays in clinical diagnostics, life science research, environmental testing, food and water evaluation, and th

Nanomaterials for Biosensors

Author : Bansi D. Malhotra
Publisher : William Andrew
ISBN 13 : 0128135158
Total Pages : 334 pages
Book Rating : 4.1/5 (281 download)

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Book Synopsis Nanomaterials for Biosensors by : Bansi D. Malhotra

Download or read book Nanomaterials for Biosensors written by Bansi D. Malhotra and published by William Andrew. This book was released on 2017-10-26 with total page 334 pages. Available in PDF, EPUB and Kindle. Book excerpt: Nanomaterials for Biosensors: Fundamentals and Applications provides a detailed summary of the main nanomaterials used in biosensing and their application. It covers recent developments in nanomaterials for the fabrication of biosensor devices for healthcare diagnostics, food freshness and bioprocessing. The various processes used for synthesis and characterization of nanostructured materials are examined, along with the design and fabrication of bioelectronic devices using nanostructured materials as building blocks. Users will find the fundamentals of the main nanomaterials used in biosensing, helping them visualize a systematic and coherent picture of how nanomaterials are used in biosensors. The book also addresses the role of bio-conjugation of nanomaterials in the construction of nano-biointerfaces for application in biosensors. Such applications, including metal nanoparticles, metal oxide nanoparticles, nanocomposites, carbon nanotubes, conducting polymers and plasmonic nanostructures in biosensing are discussed relative to each nanomaterial concerned. Finally, recent advancements in protein functionalized nanomaterials for cancer diagnostics and bio-imaging are also included. Provides a detailed study on how nanomaterials are used to enhance sensing capabilities in biosensors Explains the properties, characterization methods and preparation techniques of the nanomaterials used in biosensing Arranged in a material-by-material way, making it clear how each nanomaterial should be used

Electroanalytical Applications of Quantum Dot-Based Biosensors

Author : Bengi Uslu
Publisher : Elsevier
ISBN 13 : 0128216700
Total Pages : 476 pages
Book Rating : 4.1/5 (282 download)

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Book Synopsis Electroanalytical Applications of Quantum Dot-Based Biosensors by : Bengi Uslu

Download or read book Electroanalytical Applications of Quantum Dot-Based Biosensors written by Bengi Uslu and published by Elsevier. This book was released on 2021-05-21 with total page 476 pages. Available in PDF, EPUB and Kindle. Book excerpt: Quantum dots (QDs) are hybrid organic/inorganic nanoparticles with novel physical properties. QDs have two components: an inorganic core and an optically active coated shell. Moreover, surface coatings can be applied to QDs to modify the particle as needed for experiments. Hydrophilic coatings prevent leaking of metal cargo from the core, enhancing the solubility in biological contexts and bind molecules, such as receptor-ligands, antibodies, therapeutic, and diagnostic macromolecules for enhanced effects. Their high surface-to-volume ratio allows multiple functional groups to attach onto the surface of the particles at constant surface volume. Silicon-, gallium-, indium-, or germanium-based; cadmium-based; and carbon-based QDs have already been used in many applications, such as imaging probes for the engineering of multifunctional nanodevices. Superior properties of QDs make them an excellent system in technology and biotechnology. This book describes electroanalytical applications of QD-based nanobiosensors, including brief information about the synthesis and characterization of QDs and basics of electroanalytical methods, followed by QDs in electrochemical biomimetic sensors, QDs in microchips, inorganic materials doped QDs, QD-based electrochemical DNA biosensors, electroluminescence for biomarker analysis using aptamer-based QDs, QD-based photoelectrochemical techniques, enzyme-based nanobiosensors using QDs, QD-based electrochemical immunosensors, and QD-modified nanosensors in drug analysis. Outlines QD-based applications for drug, food, clinical, and environmental science Shows how the properties of QDs make them effective ingredients in biosensing applications Assesses the major challenges in integrating QDs in biosensing systems

Nano-inspired Biosensors for Protein Assay with Clinical Applications

Author : Genxi Li
Publisher : Elsevier
ISBN 13 : 0128150548
Total Pages : 380 pages
Book Rating : 4.1/5 (281 download)

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Book Synopsis Nano-inspired Biosensors for Protein Assay with Clinical Applications by : Genxi Li

Download or read book Nano-inspired Biosensors for Protein Assay with Clinical Applications written by Genxi Li and published by Elsevier. This book was released on 2018-08-29 with total page 380 pages. Available in PDF, EPUB and Kindle. Book excerpt: Nano-inspired Biosensors for Protein Assay with Clinical Applications introduces the latest developments in nano-inspired biosensing, helping readers understand both the fundamentals and frontiers in this rapidly advancing field. In recent decades, there has been increased interest in nano-inspired biosensors for clinical application. Proteins, e.g. antigen-antibody, tumor markers and enzymes are the most important target in disease diagnosis, and a variety of biosensing techniques and strategies have been developed for protein assay. This book brings together all the current literature on the most recent advances of protein analysis and new methodologies in designing new kinds of biosensors for clinical diagnostic use. Provides a single source of information on the latest developments in the field of biosensors for protein analysis and clinical diagnosis Focuses on biosensors fabricated with nanomaterials and nanotechnology Gives detailed methodologies for designing and fabricating nano-inspired biosensors

Heterofunctional Solid-binding Peptides for Nucleic Acid Sensing Towards the Development of Cancer Biosensors

Author : Richard Victorino Lee
Publisher :
ISBN 13 :
Total Pages : 52 pages
Book Rating : 4.:/5 (124 download)

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Book Synopsis Heterofunctional Solid-binding Peptides for Nucleic Acid Sensing Towards the Development of Cancer Biosensors by : Richard Victorino Lee

Download or read book Heterofunctional Solid-binding Peptides for Nucleic Acid Sensing Towards the Development of Cancer Biosensors written by Richard Victorino Lee and published by . This book was released on 2020 with total page 52 pages. Available in PDF, EPUB and Kindle. Book excerpt: A key challenge in designing biosensors is achieving high detection sensitivity without compromising target specificity. Field-effect transistors based on 2D solids offer enhanced sensitivity due to their atomically thin characteristics. Still, such biosensors must possess several critical attributes to detect ultra-low target concentrations within a bodily fluid: 1) Probes must be securely immobilized onto the 2D-layer substrate; 2) Optimal molecular packing of the probe must be controlled for efficient target capture; 3) Non-specific adsorption of off-target molecules must be prevented; and 4) Non-covalent functionalization of the sensing surface is favored to avoid creating surface defects that could affect sensing properties. Achieving these attributes presents several major obstacles. In the present research, these challenges are addressed to set the foundation for further development towards a versatile cancer diagnostic device. An anti-fouling graphene-binding peptide (GrBP) was used to confer passivating properties onto the sensing surface which mitigates non-specific protein adsorption, while a heterofunctional GrBP chimera bearing a peptide nucleic acid linker was used (GrBP-PNA) to immobilize DNA probes onto the sensing surface. Sensor surface functionalization, followed by subsequent target capture, was confirmed using surface plasmon resonance (SPR) and quartz crystal microbalance (QCM) analysis. Raman spectroscopy of the graphene FET device revealed a pristine surface ready functionalization in order to detect nucleic acid biomarker targets. The results provide a viable biosensing strategy for versatile nucleic acid sensing using a modular probe design. This modular approach and anti-fouling method can allow for the eventual goal of collecting miRNA expression profiles from complex biological liquids for clinical diagnosis and prognosis of various diseases.

Towards a Fibre Optic Nucleic Acid Biosensor, Thiazole Orange Derivatives as Sensitive Fluorescent Probes to Detect DNA Hybridization

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Author : Dalia Hanafi-Bagby
Publisher :
ISBN 13 :
Total Pages : pages
Book Rating : 4.:/5 (133 download)

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Book Synopsis Towards a Fibre Optic Nucleic Acid Biosensor, Thiazole Orange Derivatives as Sensitive Fluorescent Probes to Detect DNA Hybridization by : Dalia Hanafi-Bagby

Download or read book Towards a Fibre Optic Nucleic Acid Biosensor, Thiazole Orange Derivatives as Sensitive Fluorescent Probes to Detect DNA Hybridization written by Dalia Hanafi-Bagby and published by . This book was released on 1998 with total page pages. Available in PDF, EPUB and Kindle. Book excerpt: Single-stranded DNA oligonucleotides (dT20) were immobilized onto the functionalized surface of fused silica optical fibres ' via' a hexaethylene glycol linker. 1-hydroxyethyl-4-[(3-methyl-6-methoxybenzothiazole-2-ylidine)methine] quinolium bromide (TOMEHE) was synthesized and characterized by mass spectrometry and uv-visible absorption spectrophotometry. Absorption of TOMEHE in the presence of single-stranded DNA or double-stranded DNA was found to be linear (r 2 = 0.998 and 0.995, respectively) over the concentration range studied (1:60, 1:30, 1:12, 1:3 and 1:1 dye:base or base pair ratios). Hybridization events were detected using a fibre optic nucleic: acid detection scheme whereby delivery of radiation to the surface of the optical fibre was achieved by total internal reflection. Fluorescence from TOMEHE associated with the DNA membrane was coupled into the fibre and measured. The fluorescence intensity maximum of TOMEHE associated with immobilized dT20 on the surface of the optical fibre was observed to lie at 560 ± 3 nm. The wavelength of the fluorescence maximum for TOMEHE when associated with hybidized DNA on the fibre surface was not observed to be significantly different from that of single-stranded DNA. TOMEHE demonstrated a 10-fold greater fluorescence enhancement over EB when bound to double-stranded DNA, but also displayed a 2.5 fold enhancement over EB when associated with single-stranded DNA. The linearity of fluorescence for TOMEHE when bound to double-stranded DNA was poor (r2 = 0.566) for the range studied (1:60, 1:30, 1:12, 1:3 and 1:1 dye:base pair ratios), suggesting that this dye may not be suitable for incorporation into the nucleic acid detection strategy described herein.

Development and Microfabrication of Next Generation Biosensors for Nucleic Acids and Neurotransmitters

Author : Yan Cao
Publisher :
ISBN 13 :
Total Pages : 0 pages
Book Rating : 4.:/5 (138 download)

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Book Synopsis Development and Microfabrication of Next Generation Biosensors for Nucleic Acids and Neurotransmitters by : Yan Cao

Download or read book Development and Microfabrication of Next Generation Biosensors for Nucleic Acids and Neurotransmitters written by Yan Cao and published by . This book was released on 2023 with total page 0 pages. Available in PDF, EPUB and Kindle. Book excerpt: Nucleic acid-based detection can be applied broadly to detect pathogens in, for example, clinical samples, livestock, food and water. The recent COVID-19 pandemic once again emphasized the importance of point-of-care (POC) diagnosis, which is rapid, inexpensive and reliable, to defend against public health threats. Currently, nucleic acid amplification tests (NAATs) rely on polymerase chain reaction (PCR) and other amplification methods that makes them more complex, less portable, dependent on perishable reagents and less suitable as POC tests. With the goal to develop portable, rapid and inexpensive diagnostic devices, we have been working on the further evolution of the previously reported nanopore-based, nucleic acid detection platform developed at UCLA. The detection scheme utilizes charge neutral peptide nucleic acid (PNA) probes conjugated to carboxyl-functionalized microbeads for sequence-specific nucleic acid (NA) detection. The hybridization event with target NA introduces negative charge to the PNA-bead complex which results in electrophoretic mobility. The negatively charged PNA-bead complex after target NA hybridization is directed by an electric field to a nanopore of smaller diameter in a micron-thick glass membrane where it blocks at least partially the ionic current flow thereby providing a sustained current drop which serves as the detection signal. In order to demonstrate the potential broad application for detection of pathogens in clinical samples, both Neisseria gonorrhoeae and Chlamydia trachomatis spiked in human urine were detected at 10-100 CFU/mL against a background of representative bacterial flora. High sensitivities and specificities of ~95% were demonstrated and no false positives were observed with the representative bacterial flora which served as a control. An improvement of overall detection time was achieved by integrating the nanopore detector, which consists of a glass chip with an etched membrane and nanopore, with the lateral flow assay (LFA) format. The extracted NA sample flows along an LFA membrane by surface-tension driven capillary flow and hybridizes with a preloaded deposit of neutral PNA-beads. Driven by an external electric field, the negatively charged bead with hybridized NA successfully blocks the smaller nanopore integrated on the LFA chip and generates a detection signal. Therefore, the overall detection time is reduced significantly without the need for complex pumps or valves. Detection of 10 aM E. coli 16S rRNA against 10 fM P. putida 16S rRNA within 15 minutes was demonstrated successfully using the nanopore integrated into the LFA format. Further demonstrations with complex clinical samples and integration with simpler NA extraction schemes may lead to broad application for disease diagnosis at the POC.The study of neurological disorders and the deciphering of complex behaviors rely on the ability to monitor neuronal processes including inter-neuronal chemical signaling. Compared to extensively developed electrophysiological recordings, the detection of neurotransmitter release events in near-real time is far less advanced and remains limited in application. Neuroprobes with high spatiotemporal resolution would enable establishment of the relationships between neurotransmitter release events and subsequent behaviors. Our lab previously has reported glutamate (Glut) microbiosensors that can selectively detect glutamate against a background of electroactive interferents including dopamine (DA) and ascorbic acid (AA). In order to achieve better sensitivity and spatiotemporal resolution, further development could be achieved by optimizing the enzyme deposition technique and the thicknesses of the deposited enzyme layer and permselective polymer coatings. Microcontact printing ([mu]CP) has been adopted as a direct method to deposit arrays of proteins on silicon or glass substrates without compromising the activity of the proteins. By incorporating a specially designed polymer with choline oxide to create a [mu]CP "ink" led to improved choline sensor performance including a sensitivity of 639 ± 96 nA [mu]M-1 cm-2 (pH 7.4; n = 4). Guided by simulations performed with a detailed mathematical model, choline sensors were constructed with exhibited sensitivity and response time of 660 ± 40 nA [mu]M−1 cm−2 at 37°C and 0.36 ± 0.05 s respectively. With assistance of microcontact printing to selectively transfer different enzymes onto specific microelectrodes in an array on 100-um-width microprobes, a demonstration of an implantable multifunctional neural microprobe for simultaneous multi-analyte sensing and chemical delivery has been achieved.