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Languages : en
Pages : 26
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Author:
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Category :
Languages : en
Pages : 26
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Author: Benjamin Ryan Cipriany
Publisher:
ISBN:
Category :
Languages : en
Pages : 306
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Microfluidics and nanofluidics have recently emerged as analytical tools for the study of biology. These devices have enabled the miniaturization of biological sample preparation and detection methods, toward consuming less sample volume and improving the sensitivity and speed of analysis. This thesis explores methods for rapid detection and sorting of individual biomolecules within a nanofluidic channel. In these devices, constructed using thin-film processing techniques, attoliter-scale volume confinement is formed to isolate individual, fluorophore-labeled biomolecules in solution for absolute quantification. These devices enable studies of the unique attributes of each molecule, often masked in ensemble-averaged measurements. Statistical sampling of many molecules is achieved by voltage-actuated, electrokinetic flow within the nanofluidic device to precisely control molecule analysis rate and achieve high throughput single molecule detection (SMD). This nanofluidic technology is applied to epigenetic analysis, enabling the study of epigenetic modifications at a single molecule level. Viable epigenetic analysis within a nanofluidic device is demonstrated using chromatin, DNA bound with histone proteins, which is shown to remain in its native state during nanofluidic confinement and electrokinetic flow under physiologically-relevant conditions. Detection of an epigenetic modification, DNA methylation, is also demonstrated to elucidate its potential for detecting multiple epigenetic marks on an individual molecule. Subsequently, an architecture for automated, high-speed sorting of individual molecules is developed. In this architecture, digital signal processing methods are implemented in a field programmable gate array to achieve real-time SMD. An electric circuit model is developed to actuate and switch electrokinetic flow of molecules, partitioning them into branches of a bifurcated nanofluidic device. An optical system for parallel SMD is realized to experimentally validate the actuation of molecule sorting in-situ. Combined, these components are utilized in automated, fluorescence-activated sorting of individual, methylated DNA molecules, which were then collected for further analysis. This device is reconfigurable and can be generalized for application to fluorescence-activated separations of other molecule types. Finally, a study of various methods for optofluidic integration is presented. The optical properties of fused-silica, silicon nitride, polydimethylsiloxane, hydrogen silisequioxane, and chemical vapor deposited oxides are investigated to consider their use in SMD applications requiring ultra-low autofluorescence and high confinement of the optical probe volume. Findings were then applied to form an optical waveguide as an fluorescence excitation source toward the dense integration of optical and nanofluidic components.
Author: Kazuma Mawatari
Publisher: World Scientific
ISBN: 1848168012
Category : Science
Languages : en
Pages : 187
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Book Description
For the past decade, new research fields utilizing microfluidics have been formed. General micro-integration methods were proposed, and the supporting fundamental technologies were widely developed. These methodologies have made various applications in the fields of analytical and chemical synthesis, and their superior performances such as rapid, simple, and high efficient processing have been proved. Recently, the space is further downscaling to 101 103nm scale (we call the space extended-nano space). The extended-nano space located between the conventional nanotechnology (100 101nm) and microtechnology (>1 m), and the research tools are not well established. In addition, the extended-nano space is a transient space from single molecules to bulk condensed phase, and fluidics and chemistry are not unknown. For these purposes, basic methodologies were developed, and new specific phenomena in fluidics and chemistry were found. These new phenomena were applied to unique chemical operations such as concentration and ion selection. The new research fields which are now being created are quite different from those in microspace. Unique devices are also increasingly being reported. In this book, we describe the fundamental technologies for extended-nano space and show the unique liquid properties found in this space and applications for single molecule or cell analysis. The research area is very new and hence, exciting. In contrast to other specialized areas, the research fields require wide knowledge (chemistry, fluidics, mechanics, photonics, biology etc.) and state-of-the-art technologies (bottom-up and top-down fabrication for various hard and soft materials, precise fluidic control, single molecule detection methods, and particle surface modification methods etc.), which have not been not covered by conventional review papers or books. Therefore, researchers or students new to the field need a new book covering these fields including recent research topics, applications and problems to be solved in the future. Our motivation is to summarize the state-of-the-art technologies for research and demonstrate new chemistry and fluidics in extended-nano space for students and researchers in academia or industry. We also emphasize the potential large impact microfluidic technologies have on chemistry and biochemistry.
Author: Chen Wang
Publisher: Springer Science & Business Media
ISBN: 3540395024
Category : Technology & Engineering
Languages : en
Pages : 306
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Book Description
Single-molecule studies constitute a distinguishable category of focused - search in nanoscience and nanotechnology. This book is dedicated to the - troduction of recent advances on single-molecule studies. It will be illustrated that studying single molecules is both intellectually and technologically ch- lenging, and also o?ers vast potential in opening up new scienti?c frontiers. We wish to present the readers with several di?erent techniques for studying single molecules, such as electron-tunneling methods, interaction-force m- surement techniques, optical spectroscopy, plus a number of directions where further progress could be pursued. We hope the work may assist the readers, especially graduate students and those who wish to explore single molecules, to become familiarized with the pace of the progress in this ?eld and the relevant primary techniques. Due to limitation of space, we are not able to elaborate on the technical details of all of the experimental methods that are vital in single molecule studies, so introductions to only selected experimental methods are touched in the context. Since the technical details and theoretical analysis of these techniqueshavealreadybeenthoroughlycoveredinmanyliteratures,weonly provide introductions to the basic principles of the detection techniques here, and focus on their experimental achievements in the area of single-molecule studies. These techniques have proven to be highly e?ective when indep- dently used. The combinationof those techniques could lead to further - vances in the detection capabilities.
Author:
Publisher: Elsevier
ISBN: 0128164646
Category : Science
Languages : en
Pages : 404
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Book Description
Spectroscopy and Dynamics of Single Molecules: Methods and Applications reviews the most recent developments in spectroscopic methods and applications. Spectroscopic techniques are the chief experimental methods for testing theoretical models and research in this area plays an important role in stimulating new theoretical developments in physical chemistry. This book provides an authoritative insight into the latest advances in the field, highlighting new techniques, current applications, and potential future developments An ideal reference for chemists and physicists alike, Spectroscopy and Dynamics of Single Molecules: Methods and Applications is a useful guide for all those working in the research, design, or application of spectroscopic tools and techniques across a wide range of fields. - Includes the latest research on ultrafast vibrational and electronic dynamics, nonlinear spectroscopies, and single-molecule methods - Makes the content accessible to researchers in chemistry, biophysics, and chemical physics through a rigorous multi-disciplinary approach - Provides content edited by a world-renowned chemist with more than 30 years of experience in research and instruction
Author: Nan Jing
Publisher:
ISBN:
Category :
Languages : en
Pages :
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The objective of this work is to develop a micro/nanofluidic-based single molecule detection (SMD) scheme, which would allow us to inspect individual protein or protein complex study protein-protein interactions and their dynamics. This is a collaboration work with MD Anderson Cancer Center and we applied this scheme to study functions of various proteins related to cancer progression in hope to shed new light on cancer research. State-of-the-art micro/nano-fabrication technology is used to provide fused silica micro/nano-fluidic channel devices as our detection platform. Standard contact photolithography, projection photolithography and advanced electron-beam lithography are used to fabricate micro/nano-fluidic channel with width ranging from 100nm to 2[micron]. The dimensions of these miniaturized biochips are designed to ensure single molecule resolution during detection and shrinking the detection volume leads to increase in signal-to-noise ratio, which is very critical for SMD. To minimize surface adsorption of protein, a fused silica channel surface coating procedure is also developed and significantly improved the detection efficiency. A fluorescent-labeled protein sample solution is filled in the fluidic channel by capillary force, and proteins are electro-kinetically driven through the fluidic channel with external voltage source. Commercial functionalized Quantum Dots (Qdots) are used as fluorescent labels due to its various advantages over conventional organic dyes for single molecule multi-color detection application. A fluorescence correlation spectrometer system, equipped with a 375nm diode laser, 60x water immersion objective with N.A. of 1.2 and two avalanche photodiodes (APD) is implemented to excite single molecules as well as collect emitted fluorescence signals. A two-dimensional photon burst analysis technique (photon counts vs. burst width) is developed to analyze individual single molecule events. We are able to identify target protein or protein complex directly from cell lysate based on fluorescence photon counts, as well as study the dynamics of protein-protein interactions. More importantly, with this technique we are also able to assess interactions between three proteins, which cannot be done with current ensemble measurement techniques. In summary, the technique described in this work has the advantages of high sensitivity, short processing time (2-3 minutes), very small sample consumption and high resolution quantitative analysis. It could potentially revolutionize the area of protein interaction research and provides us with more clues for the future of cancer diagnostics and treatments.
Author: Joshua Edel
Publisher: Royal Society of Chemistry
ISBN: 1849734046
Category : Science
Languages : en
Pages : 326
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Book Description
The Nanoscience and Nanotechnology Series provides a comprehensive resource of books covering key topics such as the synthesis, characterisation, performance and properties of nanostructured materials and technologies and their applications.
Author: Iddo Heller
Publisher: Springer Nature
ISBN: 1071633775
Category : Science
Languages : en
Pages : 511
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Book Description
This third edition volume expands on the previous editions with new discussions on the latest techniques and developments in the field. The chapters in this book are organized into four parts, and cover topics such as optical tweezers; single-molecule fluorescence tools; atomic force microscopy; magnetic tweezers; applications to virus protein shells, unfolding of proteins, nucleic acids, motor proteins, in vivo and in vitro; and protocols to establish specific surface interactions and perform force calibration. Written in the highly successful Methods in Molecular Biology series format, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and tips on troubleshooting and avoiding known pitfalls. Cutting-edge and thorough, Single Molecule Analysis: Methods and Protocols, Third Edition is a valuable resource for all researchers who want to learn more about this exciting and still expanding field. Chapters 2, 7, 8, 9, 12, 18, and 19 are available open access under a Creative Commons Attribution 4.0 International License via link.springer.com.
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Category :
Languages : en
Pages : 11
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Despite groundbreaking potential in a broad application space, several nanofluidic phenomena remain poorly understood. Toward advancing the understanding of fluid behavior under nanoscale confinement, we developed a novel, ideal platform for fundamental molecular transport studies, in which the fluidic channel is a single carbon nanotube (CNT). CNTs offer the advantage of simple chemistry and structure, which can be synthetically tuned with nanometer precision and accurately modeled. With combined experimental and computational approaches, we demonstrated that CNT pores with 1-5 nm diameters conduct giant ionic currents that follow an unusual sublinear electrolyte concentration dependence. The large magnitude of the ionic conductance appears to originate from a strong electroosmotic flow in smooth CNT pores. First-principle simulations suggest that electroosmotic flow arises from localized negative polarization charges on carbon atoms near a potassium (K+) ion and from the strong cation-graphitic wall interactions, which drive K+ ions much closer to the wall than chlorides (Cl- ). Single-molecule translocation studies reveal that charged molecules may be distinguished from neutral species on the basis of the sign of the transient current change during their passage through the nanopore. Together with shedding light on a few controversial questions in the CNT nanofluidics area, these results may benefit LLNL's Security Mission by providing the foundation for the development of advanced single-molecule detection system for bio/chem/explosive analytes. In addition, these experimental and computational platforms can be applied to advance fundamental knowledge in other fields, from energy storage and membrane separation to superfluid physics.
Author: Yi-Heng Sen
Publisher:
ISBN:
Category :
Languages : en
Pages : 58
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This thesis focuses on characterizing and controlling the translocation of single 48.5 kbp [lambda]-DNA molecules through an artificial nanopore with the objective of enabling multiple measurements on the same molecule. This approach may enable nanopore sensors with enhanced size or charge resolution through statistical averaging over multiple detection events. Nanopores with dimensions of 200 nm x 500 nm x 5 pm connected by microfluidic channels were fabricated using soft lithography in polydimethylsiloxane (PDMS). The PDMS nanopore could successfully detect translocation events of single [lambda]DNA molecules. Factors such as applied voltage bias, DNA concentration, and dimensions of the channel were found to affect the frequency of translocation events and signal-to-noise ratio, which are critical factors for implementing multiple measurements on the same molecule with feedback control. Noise contributions from each part of the experimental apparatus and device were also characterized. Feedback control using Labview was implemented to reverse the direction of applied voltage bias upon detection of a translocation event. The direction of travel of single DNA molecules could be successfully reversed and two measurements on the same molecule were realized. This work lays the foundations for a nanofluidic device for enhanced measurement resolution through statistical averaging over multiple measurements on the same molecule.
