Download or read book Fluorescent Protein Resonance Energy Transfer for High throughput Screening written by Annalee Whitefield Nguyen and published by . This book was released on 2006 with total page 314 pages. Available in PDF, EPUB and Kindle. Book excerpt: The desire to track proteins and monitor their functions inside of living cells has driven the development of new tools and techniques capable of making these measurements. Fluorescent protein Forster resonance energy transfer (FRET) is an attractive choice for probing intracellular events, but the low efficiency of most available pairs limits the utility of the technique. To address this problem, a quantitative evolutionary strategy using fluorescence-activated cell sorting was applied to optimize a cyan-yellow fluorescent protein pair for FRET. Multiple rounds of mutation, followed by sorting of the leading edge of the cell population enabled step-wise evolution of variants with improved FRET properties. The best pair of variants, CyPet-YPet, exhibited a seven-fold improvement in dynamic range over the parental pair when measured in vitro. Additionally, the optimized pair provided significantly improved resolution of FRET signal changes in bacterial, yeast, and mammalian cells using flow cytometry. The improvements in dynamic range were verified in applications including the detection of protease activities, protein-protein interactions, and small molecules.
Download or read book Luminescence Biotechnology written by Knox Van Dyke and published by CRC Press. This book was released on 2001-12-20 with total page 621 pages. Available in PDF, EPUB and Kindle. Book excerpt: The dangers and drawbacks inherent in radioactivity-based methods along with a demonstrated and dramatic increase in sensitivity have precipitated a major shift towards luminescence measurements and visualization techniques. Their use has now spread even to traditional clinical environments, and their applications have grown from clinical assays to
Download or read book Fluorescence Based Biosensors written by and published by Academic Press. This book was released on 2012-12-31 with total page 449 pages. Available in PDF, EPUB and Kindle. Book excerpt: One of the major challenges of modern biology and medicine consists in finding means to visualize biomolecules in their natural environment with the greatest level of accuracy, so as to gain insight into their properties and behaviour in a physiological and pathological setting. This has been achieved thanks to the design of novel imaging agents, in particular to fluorescent biosensors. Fluorescence Biosensors comprise a large set of tools which are useful for fundamental purposes as well as for applications in biomedicine, drug discovery and biotechnology. These tools have been designed and engineered thanks to the combined efforts of chemists and biologists over the last decade, and developed hand in hand together with imaging technologies. This volume will convey the many exciting developments the field of fluorescent biosensors and reporters has witnessed over the recent years, from concepts to applications, including chapters on the chemistry of fluorescent probes, on technologies for monitoring protein/protein interactions and technologies for imaging biosensors in cultured cells and in vivo. Other chapters are devoted to specific examples of genetically-encoded reporters, or to protein and peptide biosensors, together with examples illustrating their application to cellular and in vivo imaging, biomedical applications, drug discovery and high throughput screening. - Contributions from leading authorities - Informs and updates on all the latest developments in the field
Download or read book Fluorescence Resonance Energy Transfer Studies of Protein Interactions written by Sarah Friede Martin and published by . This book was released on 2008 with total page 424 pages. Available in PDF, EPUB and Kindle. Book excerpt:
Download or read book FRET and FLIM Techniques written by Theodorus W. J. Gadella and published by Elsevier. This book was released on 2011-07-29 with total page 603 pages. Available in PDF, EPUB and Kindle. Book excerpt: This volume reviews the techniques Förster Resonance Energy Transfer (FRET) and Fluorescence Lifetime Imaging Microscopy (FLIM) providing researchers with step by step protocols and handy hints and tips. Both have become staple techniques in many biological and biophysical fields.
Download or read book A Practical Guide to Assay Development and High Throughput Screening in Drug Discovery written by Taosheng Chen and published by CRC Press. This book was released on 2009-12-21 with total page 291 pages. Available in PDF, EPUB and Kindle. Book excerpt: The development of suitable assays, the integration of appropriate technology, and the effective management of the essential infrastructure are all critical to the success of any high-throughput screening (HTS) endeavor. However, few scientists have the multidisciplinary experience needed to control all aspects of an HTS drug discovery project. A P
Download or read book Quantum Dot fluorescent Protein Pairs as Fluorescence Resonance Energy Transfer Pairs written by Allison Marie Dennis and published by . This book was released on 2009 with total page pages. Available in PDF, EPUB and Kindle. Book excerpt: Fluorescence resonance energy transfer (FRET)-based biosensors have been designed to fluorometrically detect everything from proteolytic activity to receptor-ligand interactions and structural changes in proteins. While a wide variety of fluorophores have demonstrated effectiveness in FRET probes, several potential sensor components are particularly notable. Semiconductor quantum dots (QDs) are attractive FRET donors because they are rather bright, exhibit high quantum yields, and their nanoparticulate structure enables the attachment of multiple acceptor molecules. Fluorescent proteins (FPs) are also of particular interest for fluorescent biosensors because design elements necessary for signal transduction, probe assembly, and device delivery and localization for intracellular applications can all be genetically incorporated into the FP polypeptide. : The studies described in this thesis elucidate the important parameters for concerted QD-FP FRET probe design. Experimental results clarify issues of FRET pair selection, probe assembly, and donor-acceptor distance for the multivalent systems. Various analysis approaches are compared and guidelines asserted based on the results. To demonstrate the effectiveness of the QD-FP FRET probe platform, a ratiometric pH sensor is presented. The sensor, which uses the intrinsic pH-sensitivity of the FP mOrange to modulate the FP/QD emission ratio, exhibits a 20-fold change in its ratiometric measurement over a physiologically interesting pH range, making it a prime candidate for intracellular imaging applications.
Download or read book Green Fluorescent Proteins written by and published by Academic Press. This book was released on 1998-10-07 with total page 424 pages. Available in PDF, EPUB and Kindle. Book excerpt: This volume is an authoritative and comprehensive treatment of the approaches and techniques used for Green Fluorescent Proteins (GFP). The primary focus of this work is on research using biological systems. The volume covers all aspects of GFP, from its expression in different organisms to specific microscopic and data analysis methods.Key Features* Only volume on Green Fluorescent Protein research* Covers all aspects of GFP* Provides specific microscopic and data analysis methods* Discusses the design and construction of GFP fusion proteins* Covers GFP expression in animals, insects, plants, and microbes* Details procedures for time lapse imaging of living cells* Explains how to implement single molecule fluorescence detection with GFP* Discusses dual label GFP strategies for multicolor fluorescence* Presents fluorescence resonance energy transfer methods with GFPs* Details quantitative fluorescence imaging techniques* Extensively illustrated with color photographs
Download or read book Fluorescent Proteins written by Kevin F. Sullivan and published by Elsevier. This book was released on 2007-12-14 with total page 613 pages. Available in PDF, EPUB and Kindle. Book excerpt: This new edition of Fluorescent Proteins presents current applications of autofluorescent proteins in cell and molecular biology authored by researchers from many of the key laboratories in the field. Starting from a current review of the broad palette of fluorescent proteins available, several chapters focus on key autofluorescent protein variants, including spectral variants, photodynamic variants as well as chimeric FP approaches. Molecular applications are addressed in chapters that detail work with single molecules, approaches to generating protein fusions and biosensors as well as analysis of protein-protein interactions in vivo by FRET, fluorescence polarization and fluorescence cross correlation techniques. A number of approaches to in vivo dynamics are presented, including FRAP, photoactivation, and 4-dimensional microscopy. Behavior of spindle components, membrane proteins, mRNA trafficking as well as analysis of cell types in tissues and in development are detailed and provide models for a wide variety of experimental approaches. In addition, several chapters deal directly with the computational issues involved in processing multidimensional image data and using fluorescent imaging to probe cellular behavior with quantitative modeling. This volume brings together the latest perspective and techniques on fluorescent proteins and will be an invaluable reference in a wide range of laboratories.
Download or read book FRET F rster Resonance Energy Transfer written by Igor L. Medintz and published by John Wiley & Sons. This book was released on 2013-10-17 with total page 824 pages. Available in PDF, EPUB and Kindle. Book excerpt: FRET – Förster Resonance Energy Transfer Meeting the need for an up-to-date and detailed primer on all aspects of the topic, this ready reference reflects the incredible expansion in the application of FRET and its derivative techniques over the past decade, especially in the biological sciences. This wide diversity is equally mirrored in the range of expert contributors. The book itself is clearly subdivided into four major sections. The first provides some background, theory, and key concepts, while the second section focuses on some common FRET techniques and applications, such as in vitro sensing and diagnostics, the determination of protein, peptide and other biological structures, as well as cellular biosensing with genetically encoded fluorescent indicators. The third section looks at recent developments, beginning with the use of fluorescent proteins, followed by a review of FRET usage with semiconductor quantum dots, along with an overview of multistep FRET. The text concludes with a detailed and greatly updated series of supporting tables on FRET pairs and Förster distances, together with some outlook and perspectives on FRET. Written for both the FRET novice and for the seasoned user, this is a must-have resource for office and laboratory shelves.
Download or read book Make Life Visible written by Yoshiaki Toyama and published by Springer Nature. This book was released on 2019-10-02 with total page 292 pages. Available in PDF, EPUB and Kindle. Book excerpt: This open access book describes marked advances in imaging technology that have enabled the visualization of phenomena in ways formerly believed to be completelyimpossible. These technologies have made major contributions to the elucidation of the pathology of diseases as well as to their diagnosis and therapy. The volume presents various studies from molecular imaging to clinical imaging. It also focuses on innovative, creative, advanced research that gives full play to imaging technology inthe broad sense, while exploring cross-disciplinary areas in which individual research fields interact and pursuing the development of new techniques where they fuse together. The book is separated into three parts, the first of which addresses the topic of visualizing and controlling molecules for life. Th e second part is devoted to imaging of disease mechanisms, while the final part comprises studies on the application of imaging technologies to diagnosis and therapy. Th e book contains the proceedings of the 12th Uehara International Symposium 2017, “Make Life Visible” sponsored by the Uehara Memorial Foundation and held from June 12 to 14, 2017. It is written by leading scientists in the field and is an open access publication under a CC BY 4.0 license.
Download or read book High Throughput Screening written by William P. Janzen and published by Springer Science & Business Media. This book was released on 2008-02-05 with total page 269 pages. Available in PDF, EPUB and Kindle. Book excerpt: In High Throughput Screening, leading scientists and researchers expert in molecular discovery explain the diverse technologies and key techniques used in HTS and demonstrate how they can be applied generically. Writing to create precisely the introductory guidebook they wish had been available when they started in HTS, these expert seasoned authors illuminate the HTS process with richly detailed tutorials on the biological techniques involved, the management of compound libraries, and the automation and engineering approaches needed. Extensive discussions provide readers with all those key elements of pharmacology, molecular biology, enzymology, and biochemistry that will ensure the identification of suitable targets and screens, and detail the technology necessary to mine millions of data points for meaningful knowledge.
Download or read book Fluorescence Microscopy written by Ulrich Kubitscheck and published by John Wiley & Sons. This book was released on 2017-03-27 with total page 508 pages. Available in PDF, EPUB and Kindle. Book excerpt: Zu dem Thema gibt es viele Publikationen, die von Experten für Experten geschrieben wurden. Dieses Buch wendet sich insbesondere an Studenten höherer Semester und Forscher, denen das Hintergrundwissen der Physik fehlt, um neuartige Verfahren der Fluoreszenzmikroskopie zu verstehen. Die zweite Auflage wartet mit neuen Kapiteln und einer erweiterten Einführung auf. Der Schwerpunkt liegt auf der hochauflösenden und Einzelmolekül-Mikroskopie. Jedes Kapitel wurde von einem anerkannten Experten des Fachgebiets geschrieben und sorgfältig überarbeitet, um so die Entwicklungen der letzten Jahre wiederzugeben.
Download or read book Inhibitors of Protein Protein Interactions written by Ali Tavassoli and published by Royal Society of Chemistry. This book was released on 2020-12-07 with total page 357 pages. Available in PDF, EPUB and Kindle. Book excerpt: Protein-protein interactions (PPI) are at the heart of the majority of cellular processes, and are frequently dysregulated or usurped in disease. Given this central role, the inhibition of PPIs has been of significant interest as a means of treating a wide variety of diseases. However, there are inherent challenges in developing molecules capable of disrupting the relatively featureless and large interfacial areas involved. Despite this, there have been a number of successes in this field in recent years using both traditional drug discovery approaches and innovative, interdisciplinary strategies using novel chemical scaffolds. This book comprehensively covers the various aspects of PPI inhibition, encompassing small molecules, peptidomimetics, cyclic peptides, stapled peptides and macrocycles. Illustrated throughout with successful case studies, this book provides a holistic, cutting-edge view of the subject area and is ideal for chemical biologists and medicinal chemists interested in developing PPI inhibitors.
Download or read book Principles of Fluorescence Spectroscopy written by Joseph R. Lakowicz and published by Springer Science & Business Media. This book was released on 2013-11-11 with total page 487 pages. Available in PDF, EPUB and Kindle. Book excerpt: 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.
Download or read book Directed Enzyme Evolution written by Frances H. Arnold and published by Springer Science & Business Media. This book was released on 2008-02-02 with total page 381 pages. Available in PDF, EPUB and Kindle. Book excerpt: Directed evolution comprises two distinct steps that are typically applied in an iterative fashion: (1) generating molecular diversity and (2) finding among the ensemble of mutant sequences those proteins that perform the desired fu- tion according to the specified criteria. In many ways, the second step is the most challenging. No matter how cleverly designed or diverse the starting library, without an effective screening strategy the ability to isolate useful clones is severely diminished. The best screens are (1) high throughput, to increase the likelihood that useful clones will be found; (2) sufficiently sen- tive (i. e. , good signal to noise) to allow the isolation of lower activity clones early in evolution; (3) sufficiently reproducible to allow one to find small improvements; (4) robust, which means that the signal afforded by active clones is not dependent on difficult-to-control environmental variables; and, most importantly, (5) sensitive to the desired function. Regarding this last point, almost anyone who has attempted a directed evolution experiment has learned firsthand the truth of the dictum “you get what you screen for. ” The protocols in Directed Enzyme Evolution describe a series of detailed p- cedures of proven utility for directed evolution purposes. The volume begins with several selection strategies for enzyme evolution and continues with assay methods that can be used to screen enzyme libraries. Genetic selections offer the advantage that functional proteins can be isolated from very large libraries s- ply by growing a population of cells under selective conditions.
Download or read book Fluorescent and Luminescent Probes for Biological Activity written by W. T. Mason and published by Elsevier. This book was released on 1999-04-16 with total page 697 pages. Available in PDF, EPUB and Kindle. Book excerpt: The use of fluorescent and luminescent probes to measure biological function has increased dramatically since publication of the First Edition due to their improved speed, safety, and power of analytical approach. This eagerly awaited Second Edition, also edited by Bill Mason, contains 19 new chapters and over two thirds new material, and is a must for all life scientists using optical probes.The contents include discussion of new optical methodologies for detection of proteins, DNA and other molecules, as well as probes for ions, receptors, cellular components, and gene expression. Emerging and advanced technologies for probe detection such as confocal laser scanning microscopy are also covered. This book will be essential for those embarking on work in the field or using new methods to enhance their research.TOPICS COVERED:* Single and multiphoton confocal microscopy* Applications of green fluorescent protein and chemiluminescent reporters to gene expression studies* Applications of new optical probes for imaging proteins in gels * Probes and detection technologies for imaging membrane potential in live cells* Use of optical probes to detect microorganisms* Raman and confocal raman microspectroscopy* Fluorescence lifetime imaging microscopy* Digital CCD cameras and their application in biological microscopy
