Download and read online Single molecule Techniques in PDF and EPUB As molecular and cellular biologists move toward nano-techniques for performing experiments on single molecules rather than on populations of molecules, a comprehensive manual on how (and why) to carry out such experiments is needed. Single-Molecule Techniques: A Laboratory Manualfills this requirement – it is the first to take researchers who know nothing about single-molecule analyses to the point where they can successfully design and execute appropriate experiments. Geared toward research scientists in structural and molecular biology, biochemistry, and biophysics, the manual will be useful to all who are interested in observing, manipulating, and elucidating the molecular mechanisms and discrete properties of macromolecules. Techniques range from in vivo and in vitro fluorescent-based methods to the use of atomic force microscopy, optical and magnetic tweezers, and nanopores. The book is edited by Paul R. Selvin and Taekjip Ha, two pioneers in the field of experimental biophysics who have made significant contributions to the development and application of single-molecule technologies.
Download and read online Single Molecule Biology in PDF and EPUB Single molecule techniques, including single molecule fluorescence, optical tweezers, and scanning probe microscopy, allow for the manipulation and measurement of single biological molecules within a live cell or in culture. These approaches, amongst the most exciting tools available in biology today, offer powerful new ways to elucidate biological function, both in terms of revealing mechanisms of action on a molecular level as well as tracking the behaviour of molecules in living cells. This book provides the first complete and authoritative treatment of this rapidly emerging field, explicitly from a biological perspective. The contents are organized by biological system or molecule. Each chapter discusses insights that have been revealed about their mechanism, structure or function by single molecule techniques. Among the topics covered are enzymes, motor proteins, membrane channels, DNA, ribozymes, cytoskeletal proteins, and other key molecules of current interest. An introduction by the editor provides a concise review of key principles and an historical overview. The last section discusses applications in molecular diagnostics and drug discovery. * Organized by biological system or molecule. * Each chapter discusses insights into mechanism of action, structure, and function * Covers enzymes, motor proteins, membrane channels, DNA, ribozymes, etc. * Includes an introduction to key principles and an historical overview. * Discusses applications in molecular diagnostics and drug discovery. * Provides an expert's perspective on future developments.
Download and read online Single Cell and Single Molecule Techniques for the Analysis of the Epigenome in PDF and EPUB Epigenetic regulation is a critical biological process for the health and development of a cell. Epigenetic regulation is facilitated by covalent modifications to the underlying DNA and chromatin proteins. A fundamental understanding of these epigenetic modifications and their associated interactions at the molecular scale is necessary to explain phenomena including cellular identity, stem cell plasticity, and neoplastic transformation. It is widely known that abnormal epigenetic profiles have been linked to many diseases, most notably cancer. While the field of epigenetics has progressed rapidly with conventional techniques, significant advances remain to be made with respect to combinatoric analysis of epigenetic marks and single cell epigenetics. Therefore, in this dissertation, I will discuss our development of devices and methodologies to address these pertinent issues. First, we designed a preparatory polydimethylsiloxane (PDMS) microdevice for the extraction, purification, and stretching of human chromosomal DNA and chromatin from small cell populations down to a single cell. The valveless device captures cells by size exclusion within the micropillars, entraps the DNA or chromatin in the micropillars after cell lysis, purifies away the cellular debris, and fluorescently labels the DNA and/or chromatin all within a single reaction chamber. With the device, we achieve nearly 100% extraction efficiency of the DNA. The device is also used for in-channel immunostaining of chromatin followed by downstream single molecule chromatin analysis in nanochannels (SCAN). Second, using multi-color, time-correlated single molecule measurements in nanochannels, simultaneous coincidence detection of 2 epigenetic marks is demonstrated. Coincidence detection of 3 epigenetic marks is also established using a pulsed interleaved excitation scheme. With these two promising results, genome-wide quantification of epigenetic marks was pursued. Unfortunately, quantitative SCAN never materialized. Reasons for this, including poor signal to background, are explained in detail. Third, development of mobility-SCAN, an analytical technique for measuring and analyzing single molecules based on their fluorescent signature and their electrophoretic mobility in nanochannels is described. We use the technique to differentiate biomolecules from complex mixtures and derive parameters such as diffusion coefficients and effective charges. Finally, the device is used to detect binding interactions of various complexes similar to affinity capillary electrophoresis, but on a single molecule level. Fourth, we conclude by briefly discussing SCAN-sort, a technique to sort individual chromatin molecules based on their fluorescent emissions for further downstream analysis such as DNA sequencing. We demonstrate a 2-fold enrichment of chromatin from sorting and discuss possible system modifications for better performance in the future.
Download and read online Single Molecule Cellular Biophysics in PDF and EPUB Indispensable textbook for undergraduate students in the physical and life sciences, unravelling the inner workings of the cell.
Download and read online Single Molecule Enzymology Fluorescence Based and High Throughput Methods in PDF and EPUB Single-Molecule Enzymology, Part A, the latest volume in the Methods in Enzymology series, continues the legacy of this premier serial with quality chapters authored by leaders in the field. This volume covers research methods in single-molecule enzymology, and includes sections on such topics as force-based and hybrid approaches, fluorescence, high-throughput sm enzymology, nanopores, and tethered particle motion. Continues the legacy of this premier serial with quality chapters authored by leaders in the field Covers research methods in single-molecule enzymology Contains sections on such topics as force-based and hybrid approaches, fluorescence, high-throughput sm enzymology, nanopores, and tethered particle motion
Download and read online Single molecule Studies of Proteins in PDF and EPUB In Single Molecule Studies of Proteins, expert researchers discuss the successful application of single-molecule techniques to a wide range of biological events, such as the imaging and mapping of cell surface receptors, the analysis of the unfolding and folding pathways of single proteins, the analysis interaction forces between biomolecules, the study of enzyme catalysis or the visualization of molecular motors in action. The chapters are aimed at established investigators and post-doctoral researchers in the life sciences wanting to pursue research in the various areas in which single-molecule approaches are important; this volume also remains accessible to advanced graduate students seeking similar research goals.
Download and read online Single Molecule Enzymology Nanomechanical Manipulation and Hybrid Methods in PDF and EPUB Single-Molecule Enzymology, Part B, the latest volume in the Methods in Enzymology series, continues the legacy of this premier serial with quality chapters authored by leaders in the field. This volume covers research methods in single-molecule enzymology, and includes sections on such topics as force-based and hybrid approaches, fluorescence, high-throughput sm enzymology, and nanopore and tethered particle motion. Continues the legacy of this premier serial with quality chapters authored by leaders in the field Covers research methods in single-molecule enzymology Contains sections on such topics as force-based and hybrid approaches, fluorescence, high-throughput sm enzymology, and nanopore and tethered particle motion
Download and read online Single Molecule Dynamics in Life Science in PDF and EPUB Yanagida (Osaka University Graduate School of Frontier Biosciences) and Ishii (Core Research for Evolutional Science and Technology, Japan Science and Technology Agency) gather pioneers in the field of single molecule science to show how to set up and interpret a single molecule experiment. Following an introduction to single molecule measurements and enzymology, contributors consider molecular motors and mechanical properties, then move on to applications. Detailed discussions of studies on protein enzymes, ribozymes, and nucleic acids are also included. Some chapter topics include imaging and molecular motors, signal transduction across the plasma membrane, single-molecule tracking of quantum dot liganded epidermal growth factor, single- molecule FRET studies of helicases and Holliday junctions, and high- speed atomic force microscopy for nano-visualization of biomolecular processes. B&w and color images and illustrations are included. Annotation ©2009 Book News, Inc., Portland, OR --From booknews.com.
Download and read online Biology at the Single Molecule Level in PDF and EPUB This is the first book solely devoted to single-molecule biochemistry and molecular biology. Authors were selected on the basis of their contribution to this new and exciting field, and were asked to focus more on the biological problems that can be approached using single-molecule techniques rather than on the techniques per se. It is thought that such techniques will eventually dominate the physical characterization of biologically important macromolecules.
Download and read online Single Molecule Electronics in PDF and EPUB This book presents a multidisciplinary approach to single-molecule electronics. It includes a complete overview of the field, from the synthesis and design of molecular candidates to the prevalent experimental techniques, complemented by a detailed theoretical description. This all-inclusive strategy provides the reader with the much-needed perspective to fully understand the far-reaching ramifications of single-molecule electronics. In addition, a number of state-of-the-art topics are discussed, including single-molecule spectro-electrical methods, electrochemical DNA sequencing technology, and single-molecule chemical reactions. As a result of this integrative effort, this publication may be used as an introductory textbook to both graduate and advanced undergraduate students, as well as researchers with interests in single-molecule electronics, organic electronics, surface science, and nanoscience.
Download and read online Correlation Force Spectroscopy for Single Molecule Measurements in PDF and EPUB This thesis addresses the development of a new force spectroscopy tool, correlation force spectroscopy (CFS) for the measurement of the properties of very small volumes of material (molecular to μm3) at kHz-MHz frequency range. CFS measures the simultaneous thermal fluctuations of two closely-spaced atomic force microscopy (AFM) cantilevers. CFS then calculates the cross-correlation in the thermal fluctuations that gives the mechanical properties of the matter that spans the gap of the two cantilevers. The book also discusses development of CFS, its advantages over AFM, and its application in single molecule force spectroscopy and micro-rheology.
Download and read online Using Single Molecule Techniques to Determine the Mechanism of DNA Topology Simplification by Type IIA Topoisomerases in PDF and EPUB
Download and read online Single Molecule Tools Part A Fluorescence Based Approaches in PDF and EPUB Single molecule tools have begun to revolutionize the molecular sciences, from biophysics to chemistry to cell biology. They hold the promise to be able to directly observe previously unseen molecular heterogeneities, quantitatively dissect complex reaction kinetics, ultimately miniaturize enzyme assays, image components of spatially distributed samples, probe the mechanical properties of single molecules in their native environment, and "just look at the thing" as anticipated by the visionary Richard Feynman already half a century ago. Single Molecule Tools, Part A: Fluorescence Based Approaches captures a snapshot of this vibrant, rapidly expanding field, presenting articles from pioneers in the field intended to guide both the newcomer and the expert through the intricacies of getting single molecule tools. Includes time-tested core methods and new innovations applicable to any researcher employing single molecule tools Methods included are useful to both established researchers and newcomers to the field Relevant background and reference information given for procedures can be used as a guide to developing protocols in a number of disciplines
Download and read online Handbook of Single Molecule Biophysics in PDF and EPUB This handbook describes experimental techniques to monitor and manipulate individual biomolecules, including fluorescence detection, atomic force microscopy, and optical and magnetic trapping. It includes single-molecule studies of physical properties of biomolecules such as folding, polymer physics of protein and DNA, enzymology and biochemistry, single molecules in the membrane, and single-molecule techniques in living cells.
Download and read online Handbook of Single Molecule Fluorescence Spectroscopy in PDF and EPUB This is a practical introduction to single molecule fluorescence experiments, the analysis of the data, and applications of the techniques to the study of biological structure and function.