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.
The topics range from single molecule experiments in quantum optics and solid-state physics to analogous investigations in physical chemistry and biophysics.
Geared towards research scientists in structural and molecular biology, biochemistry, and biophysics, this manual will be useful to all who are interested in observing, manipulating and elucidating the molecular mechanisms and discrete properties of macromolecules.
This book reviews recently developed theoretical and numerical approaches to deal with optical and mechanical signals from individual molecules. The character of data generated by single molecules, and more generally by single nano-objects, qualitatively differs from those obtained in conventional experiments on large ensembles of molecules. Fluctuations, randomness and irreproducibility are central to single-molecule measurements, and the specific methods required to extract reliable and statistically relevant information from them are presented here. With contributions mainly from participants of the ?Theory, Modeling and Evaluation of Single-Molecule Measurements? workshop held in Leiden, the Netherlands, on April 16-20, 2007, this book is an authoritative compendium on the subject.
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.
This important volume contains selected papers and extensive commentaries on laser trapping and manipulation of neutral particles using radiation pressure forces. Such techniques apply to a variety of small particles, such as atoms, molecules, macroscopic dielectric particles, living cells, and organelles within cells. These optical methods have had a revolutionary impact on the fields of atomic and molecular physics, biophysics, and many aspects of nanotechnology.In atomic physics, the trapping and cooling of atoms down to nanokelvins and even picokelvin temperatures are possible. These are the lowest temperatures in the universe. This made possible the first demonstration of Bose-Einstein condensation of atomic and molecular vapors. Some of the applications are high precision atomic clocks, gyroscopes, the measurement of gravity, cryptology, atomic computers, cavity quantum electrodynamics and coherent atom lasers.A major application in biophysics is the study of the mechanical properties of the many types of motor molecules, mechanoenzymes, and other macromolecules responsible for the motion of organelles within cells and the locomotion of entire cells. Unique in vitro and in vivo assays study the driving forces, stepping motion, kinetics, and efficiency of these motors as they move along the cell's cytoskeleton. Positional and temporal resolutions have been achieved, making possible the study of RNA and DNA polymerases, as they undergo their various copying, backtracking, and error correcting functions on a single base pair basis.Many applications in nanotechnology involve particle and cell sorting, particle rotation, microfabrication of simple machines, microfluidics, and other micrometer devices. The number of applications continues to grow at a rapid rate.The author is the discoverer of optical trapping and optical tweezers. With his colleagues, he first demonstrated optical levitation, the trapping of atoms, and tweezer trapping and manipulation of living cells and biological particles.This is the only review volume covering the many fields of optical trapping and manipulation. The intention is to provide a selective guide to the literature and to teach how optical traps really work.
This updated and up-to-date version of the first edition continues with the really interesting stuff to spice up a standard biophysics and biophysical chemistry course. All relevant methods used in current cutting edge research including such recent developments as super-resolution microscopy and next-generation DNA sequencing techniques, as well as industrial applications, are explained. The text has been developed from a graduate course taught by the author for several years, and by presenting a mix of basic theory and real-life examples, he closes the gap between theory and experiment. The first part, on basic biophysical chemistry, surveys fundamental and spectroscopic techniques as well as biomolecular properties that represent the modern standard and are also the basis for the more sophisticated technologies discussed later in the book. The second part covers the latest bioanalytical techniques such as the mentioned super-resolution and next generation sequencing methods, confocal fluorescence microscopy, light sheet microscopy, two-photon microscopy and ultrafast spectroscopy, single molecule optical, electrical and force measurements, fluorescence correlation spectroscopy, optical tweezers, quantum dots and DNA origami techniques. Both the text and illustrations have been prepared in a clear and accessible style, with extended and updated exercises (and their solutions) accompanying each chapter. Readers with a basic understanding of biochemistry and/or biophysics will quickly gain an overview of cutting edge technology for the biophysical analysis of proteins, nucleic acids and other biomolecules and their interactions. Equally, any student contemplating a career in the chemical, pharmaceutical or bio-industry will greatly benefit from the technological knowledge presented. Questions of differing complexity testing the reader's understanding can be found at the end of each chapter with clearly described solutions available on the Wiley-VCH textbook homepage under: www.wiley-vch.de/textbooks
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.
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.
