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 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 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 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 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 Single Molecule Chemistry and Physics in PDF and EPUB 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.
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.
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 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 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 Particle Tracking and Single Molecule Energy Transfer in PDF and EPUB Closing a gap in the literature, this handbook gathers all the information on single particle tracking and single molecule energy transfer. It covers all aspects of this hot and modern topic, from detecting virus entry to membrane diffusion, and from protein folding using spFRET to coupled dye systems, as well recent achievements in the field. Throughout, the first-class editors and top international authors present content of the highest quality, making this a must-have for physical chemists, spectroscopists, molecular physicists and biochemists.
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 Advances in Chemical Physics Volume 146 in PDF and EPUB Discover the experimental and theoretical developments in optical single-molecule spectroscopy that are changing the ways we think about molecules and atoms The Advances in Chemical Physics series provides the chemical physics field with a forum for critical, authoritative evaluations of advances in every area of the discipline. This latest volume explores the advent of optical single-molecule spectroscopy, and how atomic force microscopy has empowered novel experiments on individual biomolecules, opening up new frontiers in molecular and cell biology and leading to new theoretical approaches and insights. Organized into two parts—one experimental, the other theoretical—this volume explores advances across the field of single-molecule biophysics, presenting new perspectives on the theoretical properties of atoms and molecules. Single-molecule experiments have provided fresh perspectives on questions such as how proteins fold to specific conformations from highly heterogeneous structures, how signal transductions take place on the molecular level, and how proteins behave in membranes and living cells.This volume is designed to further contribute to the rapid development of single-molecule biophysics research. Filled with cutting-edge research reported in a cohesive manner not found elsewhere in the literature, each volume of the Advances in Chemical Physics series serves as the perfect supplement to any advanced graduate class devoted to the study of chemical physics.