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Author: Carol A. Roach
Publisher:
ISBN: 9781124480015
Category : Emission spectroscopy
Languages : en
Pages :
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Book Description
Time resolved emission spectroscopy experiments have numerous applications within the physical sciences. For example, intrinsic and extrinsic fluorescent molecules are used in chemical and biological assays, monitoring of reactions, and imaging of structures. The need to study chemical and biological systems of greater complexity with fluorescent molecules has required the development of more sophisticated time resolved emission spectroscopic instruments, experimental techniques, and data analysis methods. Current research continues to provide new information on fundamental principles and properties of fluorescent molecules, information that is critical to proper interpretation of data generated by these more complex experiments. Chapter 4 of this dissertation discusses the development of a new instrument capable of performing full-spectrum fluorescence correlation spectroscopy. In Chapter 1 the advantages of full-spectrum fluorescence correlation spectroscopy, as well as proper interpretation of tryptophan fluorescence performed in Chapter 5, are discussed with conclusions, limitations, and concepts for future experiments presented in Chapter 6. Chapters 2 and 3 provide a background on fluorescence principles and multivariate analysis techniques as they apply to full-spectrum fluorescence correlation spectroscopy, and experiments involving tryptophan fluorescence interpretation that are presented in Chapter 5. For the first time, full spectra are collected during the course of fluorescence correlation spectroscopy measurements, allowing multivariate analysis to be applied to the data. Measurement of instrumental parameters for full-spectrum fluorescence correlation spectroscopy such as observation volume size, two-photon excitation capabilities, concentration limitations, and the range of observable diffusion coefficients are performed and compared to traditional fluorescence correlation spectroscopy instruments. An experimental application of the instrument to fluorescently labeled polystyrene particle diffusion in a polymer solution with a second fluorescent dye is shown. Separation of the two overlapped fluorescence spectra in this mixture by multivariate analysis is performed and diffusion coefficients are calculated. The results show that spectral resolution obtained by multivariate analysis of data collected with this novel instrument provides a method to reduce cross-talk in multicolor fluorescence correlation spectroscopy experiments. Experiments involving photokinetic matrix decomposition of time resolved emission decay matrices collected for the amino acid tryptophan and peptide mastoparan-X are performed. Tryptophan is a microenvironment sensitive fluorescent molecule that may naturally occur in proteins. The microenvironment sensitivity of tryptophan has been exploited to imply structural changes of proteins in time resolved fluorescence experiments, however tryptophan fluorescence photokinetics are complex. A controversy exists on the proper interpretation and use of tryptophan fluorescence, centered on the origin of the fluorescence from ground-state heterogeneity or excited-state reactions of the molecule. Photokinetic matrix decomposition of time resolved emission decay data has previously been able to differentiate between ground-state heterogeneity and excited state reaction photokinetics within other microenvironment sensitive fluorescent molecules, and is applied in this dissertation to the study of tryptophan fluorescence. Data collected with tryptophan and mastoparan-X suggest that ground-state heterogeneity is the source of complex tryptophan fluorescence decay in proteins.
Author: Carol A. Roach
Publisher:
ISBN: 9781124480015
Category : Emission spectroscopy
Languages : en
Pages :
Get Book Here
Book Description
Time resolved emission spectroscopy experiments have numerous applications within the physical sciences. For example, intrinsic and extrinsic fluorescent molecules are used in chemical and biological assays, monitoring of reactions, and imaging of structures. The need to study chemical and biological systems of greater complexity with fluorescent molecules has required the development of more sophisticated time resolved emission spectroscopic instruments, experimental techniques, and data analysis methods. Current research continues to provide new information on fundamental principles and properties of fluorescent molecules, information that is critical to proper interpretation of data generated by these more complex experiments. Chapter 4 of this dissertation discusses the development of a new instrument capable of performing full-spectrum fluorescence correlation spectroscopy. In Chapter 1 the advantages of full-spectrum fluorescence correlation spectroscopy, as well as proper interpretation of tryptophan fluorescence performed in Chapter 5, are discussed with conclusions, limitations, and concepts for future experiments presented in Chapter 6. Chapters 2 and 3 provide a background on fluorescence principles and multivariate analysis techniques as they apply to full-spectrum fluorescence correlation spectroscopy, and experiments involving tryptophan fluorescence interpretation that are presented in Chapter 5. For the first time, full spectra are collected during the course of fluorescence correlation spectroscopy measurements, allowing multivariate analysis to be applied to the data. Measurement of instrumental parameters for full-spectrum fluorescence correlation spectroscopy such as observation volume size, two-photon excitation capabilities, concentration limitations, and the range of observable diffusion coefficients are performed and compared to traditional fluorescence correlation spectroscopy instruments. An experimental application of the instrument to fluorescently labeled polystyrene particle diffusion in a polymer solution with a second fluorescent dye is shown. Separation of the two overlapped fluorescence spectra in this mixture by multivariate analysis is performed and diffusion coefficients are calculated. The results show that spectral resolution obtained by multivariate analysis of data collected with this novel instrument provides a method to reduce cross-talk in multicolor fluorescence correlation spectroscopy experiments. Experiments involving photokinetic matrix decomposition of time resolved emission decay matrices collected for the amino acid tryptophan and peptide mastoparan-X are performed. Tryptophan is a microenvironment sensitive fluorescent molecule that may naturally occur in proteins. The microenvironment sensitivity of tryptophan has been exploited to imply structural changes of proteins in time resolved fluorescence experiments, however tryptophan fluorescence photokinetics are complex. A controversy exists on the proper interpretation and use of tryptophan fluorescence, centered on the origin of the fluorescence from ground-state heterogeneity or excited-state reactions of the molecule. Photokinetic matrix decomposition of time resolved emission decay data has previously been able to differentiate between ground-state heterogeneity and excited state reaction photokinetics within other microenvironment sensitive fluorescent molecules, and is applied in this dissertation to the study of tryptophan fluorescence. Data collected with tryptophan and mastoparan-X suggest that ground-state heterogeneity is the source of complex tryptophan fluorescence decay in proteins.
Author:
Publisher:
ISBN:
Category : Medicine
Languages : en
Pages : 1060
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Book Description
Author: Jan F. Rabek
Publisher: CRC Press
ISBN: 9780849348440
Category : Science
Languages : en
Pages : 332
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Book Description
The purpose of this 4-volume book is to examine some of the applications of lasers in polymer science and technology. Now available for the first time, up-to-date information on this fascinating subject is compiled and presented in compact form. This book focuses on current research and developments in the application of lasers in polymer and biopolymer chemistry. It includes experimental and theoretical details, apparatus, techniques, and applications. This book is a useful source for researchers, students, polymer chemists, and physicists involved in this astonishing field of high technology.
Author:
Publisher: Academic Press
ISBN: 0128032251
Category : Science
Languages : en
Pages : 3716
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Book Description
This third edition of the Encyclopedia of Spectroscopy and Spectrometry, Three Volume Set provides authoritative and comprehensive coverage of all aspects of spectroscopy and closely related subjects that use the same fundamental principles, including mass spectrometry, imaging techniques and applications. It includes the history, theoretical background, details of instrumentation and technology, and current applications of the key areas of spectroscopy. The new edition will include over 80 new articles across the field. These will complement those from the previous edition, which have been brought up-to-date to reflect the latest trends in the field. Coverage in the third edition includes: Atomic spectroscopy Electronic spectroscopy Fundamentals in spectroscopy High-Energy spectroscopy Magnetic resonance Mass spectrometry Spatially-resolved spectroscopic analysis Vibrational, rotational and Raman spectroscopies The new edition is aimed at professional scientists seeking to familiarize themselves with particular topics quickly and easily. This major reference work continues to be clear and accessible and focus on the fundamental principles, techniques and applications of spectroscopy and spectrometry. Incorporates more than 150 color figures, 5,000 references, and 300 articles for a thorough examination of the field Highlights new research and promotes innovation in applied areas ranging from food science and forensics to biomedicine and health Presents a one-stop resource for quick access to answers and an in-depth examination of topics in the spectroscopy and spectrometry arenas
Author: R. Cundall
Publisher: Springer Science & Business Media
ISBN: 1475716346
Category : Science
Languages : en
Pages : 767
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Book Description
At the time that the editors conceived the idea of trying to organize the meeting on which the contents of this volume are based and which became, in March 1980, a NATO Advanced Study Institute, the techniques of time-resolved fluorescence spectroscopy, in both the nanosecond and sub-nanosecond time-domains, might reasonably have been said to be coming of age, both in their execution and in the analysis and interpretation of the results obtained. These techniques, then as now, comprised mainly a number of pulse methods using laser, flash-lamp or, most recently, synchrotron radiation. In addition, significant developments in the more classical phase approach had also rendered that method popular, utilizing either modulation of an otherwise continuous source or, again recently, the ultra-rapid pulse rate attainable with a synchrotron source. In general terms, time-resolved fluorescence studies are capable, under appropriate conditions, of supplying direct kinetic information on both photophysics and various aspects of molecular, macromolecular and supramolecular structure and dynamics. The nanosecond and sub-nanosecond time-scales directly probed render these techniques particularly appropriate in studying relaxation and fluctuation processes in macromolecules, particularly biopolymers (e. g. proteins, nucleic acids), in supramolecular assemblies such as cell membranes, and in a variety of relatively simpler model systems.
Author: Joseph R. Lakowicz
Publisher: Springer Science & Business Media
ISBN: 146157658X
Category : Science
Languages : en
Pages : 487
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Book Description
Fluorescence methods are being used increasingly in biochemical, medical, and chemical research. This is because of the inherent sensitivity of this technique. and the favorable time scale of the phenomenon of fluorescence. 8 Fluorescence emission occurs about 10- sec (10 nsec) after light absorp tion. During this period of time a wide range of molecular processes can occur, and these can effect the spectral characteristics of the fluorescent compound. This combination of sensitivity and a favorable time scale allows fluorescence methods to be generally useful for studies of proteins and membranes and their interactions with other macromolecules. This book describes the fundamental aspects of fluorescence. and the biochemical applications of this methodology. Each chapter starts with the -theoreticalbasis of each phenomenon of fluorescence, followed by examples which illustrate the use of the phenomenon in the study of biochemical problems. The book contains numerous figures. It is felt that such graphical presentations contribute to pleasurable reading and increased understand ing. Separate chapters are devoted to fluorescence polarization, lifetimes, quenching, energy transfer, solvent effects, and excited state reactions. To enhance the usefulness of this work as a textbook, problems are included which illustrate the concepts described in each chapter. Furthermore, a separate chapter is devoted to the instrumentation used in fluorescence spectroscopy. This chapter will be especially valuable for those perform ing or contemplating fluorescence measurements. Such measurements are easily compromised by failure to consider a number of simple principles.
Author: David G. Cameron
Publisher:
ISBN:
Category : Science
Languages : en
Pages : 686
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Author:
Publisher:
ISBN:
Category : Aeronautics
Languages : en
Pages : 692
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Author:
Publisher: Academic Press
ISBN: 0080957188
Category : Science
Languages : en
Pages : 3533
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Book Description
Biophysics is a rapidly-evolving interdisciplinary science that applies theories and methods of the physical sciences to questions of biology. Biophysics encompasses many disciplines, including physics, chemistry, mathematics, biology, biochemistry, medicine, pharmacology, physiology, and neuroscience, and it is essential that scientists working in these varied fields are able to understand each other's research. Comprehensive Biophysics, Nine Volume Set will help bridge that communication gap. Written by a team of researchers at the forefront of their respective fields, under the guidance of Chief Editor Edward Egelman, Comprehensive Biophysics, Nine Volume Set provides definitive introductions to a broad array of topics, uniting different areas of biophysics research - from the physical techniques for studying macromolecular structure to protein folding, muscle and molecular motors, cell biophysics, bioenergetics and more. The result is this comprehensive scientific resource - a valuable tool both for helping researchers come to grips quickly with material from related biophysics fields outside their areas of expertise, and for reinforcing their existing knowledge. Biophysical research today encompasses many areas of biology. These studies do not necessarily share a unique identifying factor. This work unites the different areas of research and allows users, regardless of their background, to navigate through the most essential concepts with ease, saving them time and vastly improving their understanding The field of biophysics counts several journals that are directly and indirectly concerned with the field. There is no reference work that encompasses the entire field and unites the different areas of research through deep foundational reviews. Comprehensive Biophysics fills this vacuum, being a definitive work on biophysics. It will help users apply context to the diverse journal literature offering, and aid them in identifying areas for further research Chief Editor Edward Egelman (E-I-C, Biophysical Journal) has assembled an impressive, world-class team of Volume Editors and Contributing Authors. Each chapter has been painstakingly reviewed and checked for consistent high quality. The result is an authoritative overview which ties the literature together and provides the user with a reliable background information and citation resource
Author: Peterson's Guides, Inc
Publisher:
ISBN:
Category : Education
Languages : en
Pages : 628
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Book Description
