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FRET and FLIM Techniques

FRET and FLIM Techniques
  • Author : Theodorus W. J. Gadella
  • Publisher :Unknown
  • Release Date :2011-07-29
  • Total pages :560
  • ISBN : 0080915124
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Summary : 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.

Molecular Imaging

Molecular Imaging
  • Author : Ammasi Periasamy,Richard Day
  • Publisher :Unknown
  • Release Date :2011-04-28
  • Total pages :312
  • ISBN : 0080536875
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Summary : The detection and measurement of the dynamic interactions of proteins within the living cell are critical to our understanding of cell physiology and pathophysiology. With FRET microscopy and spectroscopy techniques, basic and clinical scientists can make such measurements at very high spatial and temporal resolution. But sources of background information about these tools are very limited, so this book fills an important gap. It covers both the basic concepts and theory behind the various FRET microscopy and spectroscopy techniques, and the practical aspects of using the techniques and analyzing the results. The critical tricks for obtaining a good FRET image and precisely quantitating the signals from living specimens at the nanomolecular level are explained. Valuable information about the preparation of biological samples used for FRET image analysis is also provided. The methods covered include different types of microscopy systems and detectors (wide-field, confocal, multi-photon) as well as specialized techniques such as photobleaching FRET, FLIM-FRET microscopy, spectral imaging FRET, single molecule FRET, and time and image correlation spectroscopy. Starting from the basics, the chapters guide readers through the choice of probes to be used for FRET experiments and the selection of the most suitable experimental approaches to address specific biological questions. Up-to-date, consistently organized, and well-illustrated, this unique book will be welcomed by all researchers who wish to use FRET microscopy and spectroscopy techniques.

Multiphoton Microscopy and Fluorescence Lifetime Imaging

Multiphoton Microscopy and Fluorescence Lifetime Imaging
  • Author : Karsten König
  • Publisher :Unknown
  • Release Date :2018-01-22
  • Total pages :450
  • ISBN : 9783110430073
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Summary : This monograph focuses on modern femtosecond laser microscopes for two photon imaging and nanoprocessing, on laser tweezers for cell micromanipulation as well as on fluorescence lifetime imaging (FLIM) in Life Sciences. The book starts with an introduction by Dr. Wolfgang Kaiser, pioneer of nonlinear optics and ends with the chapter on clinical multiphoton tomography, the novel high resolution imaging technique. It includes a foreword by the nonlinear microscopy expert Dr. Colin Sheppard. Contents Part I: Basics Brief history of fluorescence lifetime imaging The long journey to the laser and its use for nonlinear optics Advanced TCSPC-FLIM techniques Ultrafast lasers in biophotonics Part II: Modern nonlinear microscopy of live cells STED microscopy: exploring fluorescence lifetime gradients for super-resolution at reduced illumination intensities Principles and applications of temporal-focusing wide-field two-photon microscopy FLIM-FRET microscopy TCSPC FLIM and PLIM for metabolic imaging and oxygen sensing Laser tweezers are sources of two-photon effects Metabolic shifts in cell proliferation and differentiation Femtosecond laser nanoprocessing Cryomultiphoton imaging Part III: Nonlinear tissue imaging Multiphoton Tomography (MPT) Clinical multimodal CARS imaging In vivo multiphoton microscopy of human skin Two-photon microscopy and fluorescence lifetime imaging of the cornea Multiscale correlative imaging of the brain Revealing interaction of dyes and nanomaterials by multiphoton imaging Multiphoton FLIM in cosmetic clinical research Multiphoton microscopy and fluorescence lifetime imaging for resection guidance in malignant glioma surgery Non-invasive single-photon and multi-photon imaging of stem cells and cancer cells in mouse models Bedside assessment of multiphoton tomography

Methods in Cellular Imaging

Methods in Cellular Imaging
  • Author : Ammasi Periasamy
  • Publisher :Unknown
  • Release Date :2013-05-27
  • Total pages :448
  • ISBN : 9781461475132
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Summary : Advances in technology have revolutionized the development of light microscopy techniques in biomedical research, thus improving visualization of the microstructure of cells and tissues under physiological conditions. Fluorescence microscopy methods are non-contact and non-invasive and provide high spatial and temporal resolution that other laboratory techniques cannot. This well-illustrated book targets graduate students and scientists who are new to the state-of-the-art fluorescence microscopy techniques used in biological and clinical imaging. It explains basic concepts and imaging procedures for wide-field, confocal, multiphoton excitation, fluorescence resonance energy transfer (FRET), lifetime imaging (FLIM), spectral imaging, fluorescence recovery after photobleaching (FRAP), optical tweezers, total internal reflection, high spatial resolution atomic force microscopy (AFM), and bioluminescence imaging for gene expression. The usage of these techniques in various biological applications, including calcium, pH, membrane potential, mitochondrial signaling, protein-protein interactions under various physiological conditions, and deep tissue imaging, is clearly presented. The authors describe the approaches to selecting epifluorescence microscopy, the detectors, and the image acquisition and processing software for different biological applications. Step-by-step directions on preparing different digital formats for light microscopy images on websites are also provided.

Advanced Time-Correlated Single Photon Counting Techniques

Advanced Time-Correlated Single Photon Counting Techniques
  • Author : Wolfgang Becker
  • Publisher :Unknown
  • Release Date :2005-12-20
  • Total pages :401
  • ISBN : 3540288821
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Summary : In 1984 Desmond O’Connor and David Phillips published their comprehensive book „Time-correlated Single Photon Counting“. At that time time-correlated s- gle photon counting, or TCSPC, was used primarily to record fluorescence decay functions of dye solutions in cuvettes. From the beginning, TCSPC was an am- ingly sensitive and accurate technique with excellent time-resolution. However, acquisition times were relatively slow due to the low repetition rate of the light sources and the limited speed of the electronics of the 70s and early 80s. Moreover, TCSPC was intrinsically one-dimensional, i.e. limited to the recording of the wa- form of a periodic light signal. Even with these limitations, it was a wonderful te- nique. More than 20 years have elapsed, and electronics and laser techniques have made impressive progress. The number of transistors on a single chip has approximately doubled every 18 months, resulting in a more than 1,000-fold increase in compl- ity and speed. The repetition rate and power of pulsed light sources have increased by about the same factor.

Microscopy Techniques

Microscopy Techniques
  • Author : Jens Rietdorf
  • Publisher :Unknown
  • Release Date :2005-06-23
  • Total pages :319
  • ISBN : 3540236988
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Summary : With contributions by numerous experts

FLIM Microscopy in Biology and Medicine

FLIM Microscopy in Biology and Medicine
  • Author : Ammasi Periasamy,Robert M. Clegg
  • Publisher :Unknown
  • Release Date :2009-07-06
  • Total pages :472
  • ISBN : 1420078917
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Summary : Detecting Signals at the Single Molecule Level: Pioneering Achievements in Microscopy Recent advances have led to such remarkable improvements in fluorescence lifetime imaging microscopy’s (FLIM) capacity for contrast and sensitivity that researchers can now employ it to detect signals at the single molecule level. FLIM also offers the additional benefit of independence from fluorophore concentration and excitation intensity. Moreover, its unique sensitivity makes it an excellent reporter of conformational changes and of variations in the molecular surroundings of biological molecules. Most of this improvement and discovery have occurred during the past decade, and, to date, information that would benefit a broad range of researchers remains scattered in the literature. Edited by two of the top pioneers in the field, FLIM Microscopy in Biology and Medicine presents the fundamentals of FLIM along with a number of advanced considerations so that a wider audience can appreciate recent and potential improvements that make it such a valuable tool. New Opportunities for Biomedical Researchers... New Challenges for Microscopy Researchers Discussion sections in all the chapters clearly show the challenges for implementing FLIM for various applications. Certain chapters discuss limits on the number of photons required for highly accurate lifetime determinations, as well as the accuracy with which multiple, closely associated lifetime components can reliably be determined. Such considerations are important for the user when he or she is selecting the most advantageous method of FLIM to use for a particular application. While this book provides an introduction for those new to FLIM, it gathers a wealth of material to enhance the work of experts involved in pioneering technological improvements, as well as those research opportunities in this unique and promising area of microscopy.

Plant Hormones

Plant Hormones
  • Author : Jürgen Kleine-Vehn,Michael Sauer
  • Publisher :Unknown
  • Release Date :2016-09-13
  • Total pages :280
  • ISBN : 1493964674
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Summary : This volume aims to present a representative cross-section of modern experimental approaches relevant to Plant Hormone Biology, ranging from relatively simple physiological to highly sophisticated methods. Chapters describe physiological, developmental, microscopy-based techniques, measure hormone contents, and heterologous systems. Written in the highly successful Methods in Molecular Biology series format, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and tips on troubleshooting and avoiding known pitfalls. Authoritative and cutting-edge, Plant Hormones:Methods and Protocols, Third Edition aims to provide researchers with useful methods to advance their research.

Optical Imaging Techniques in Cell Biology

Optical Imaging Techniques in Cell Biology
  • Author : Guy Cox
  • Publisher :Unknown
  • Release Date :2006-11-06
  • Total pages :288
  • ISBN : 9781420005615
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Summary : Since the word microscopy was coined in 1656, the evolution of the instrument has had a long and convoluted history. Plagued with problems of chromatic aberration, spherical aberration, and challenges with illumination and resolution, the microscope's technical progression happened in a series of fits and starts until the late 19th century. After E

Fluorescence Lifetime Spectroscopy and Imaging

Fluorescence Lifetime Spectroscopy and Imaging
  • Author : Laura Marcu,Paul M. W. French,Daniel S. Elson
  • Publisher :Unknown
  • Release Date :2014-07-17
  • Total pages :570
  • ISBN : 9781439861684
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Summary : During the past two decades, there has been an increasing appreciation of the significant value that lifetime-based techniques can add to biomedical studies and applications of fluorescence. Bringing together perspectives of different research communities, Fluorescence Lifetime Spectroscopy and Imaging: Principles and Applications in Biomedical Dia

Integrins and Ion Channels

Integrins and Ion Channels
  • Author : Andrea Becchetti,Annarosa Arcangeli
  • Publisher :Unknown
  • Release Date :2010
  • Total pages :127
  • ISBN : 1441960732
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Summary : Interdisciplinarity is more often invoked than practised. This is hardly surprising, considering the daunting vastness of modern biology. To reach a satisfactory understanding of a complex biological system, a wide spectrum of conceptual and experimental tools must be applied at different levels, from the molecular to the cellular, tissue and organismic. We believe the multifaceted regulatory interplay between integrin receptors and ion channels offers a rich and challenging field for researchers seeking broad biological perspectives. By mediating cell adhesion to the extracellular matrix, integrins regulate many developmental processes in the widest sense (from cell choice between differentiation and proliferation, to tissue remodeling and organogenesis). Rapidly growing evidence shows that frequent communication takes place between cell adhesion receptors and channel proteins. This may occur through formation of multiprotein membrane complexes that regulate ion fluxes as well as a variety of intracellular signaling pathways. In other cases, cross talk is more indirect and mediated by cellular messengers such as G proteins. These interactions are reciprocal, in that ion channel stimulation often controls integrin activation or expression. From a functional standpoint, studying the interplay between integrin receptors and ion channels clarifies how the extracellular matrix regulates processes as disparate as muscle excitability, synaptic plasticity and lymphocyte activation, just to mention a few. The derangement of these processes has many implications for pathogenesis processes, in particular for tumor invasiveness and some cardiovascular and neurologic diseases. This book provides a general introduction to the problems and methods of this blossoming field.

Principles of Fluorescence Spectroscopy

Principles of Fluorescence Spectroscopy
  • Author : Joseph R. Lakowicz
  • Publisher :Unknown
  • Release Date :2013-04-17
  • Total pages :698
  • ISBN : 9781475730616
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Summary : `In the second edition of Principles I have attempted to maintain the emphasis on basics, while updating the examples to include more recent results from the literature. There is a new chapter providing an overview of extrinisic fluorophores. The discussion of timeresolved measurements has been expanded to two chapters. Quenching has also been expanded in two chapters. Energy transfer and anisotropy have each been expanded to three chapters. There is also a new chapter on fluorescence sensing. To enhance the usefulness of this book as a textbook, most chapters are followed by a set of problems. Sections which describe advanced topics are indicated as such, to allow these sections to be skipped in an introduction course. Glossaries are provided for commonly used acronyms and mathematical symbols. For those wanting additional informtion, the final appendix contains a list of recommended books which expand on various specialized topics.' from the author's Preface

Time-correlated single photon counting

Time-correlated single photon counting
  • Author : Desmond O'Connor
  • Publisher :Unknown
  • Release Date :2012-12-02
  • Total pages :298
  • ISBN : 9780323141444
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Summary : Time-correlated Single Photon Counting has been written in the hope that by relating the authors' experiences with a variety of different single photon counting systems, they may provide a useful service to users and potential users of this formidably sensitive technique. Of all the techniques available to obtain information on the rates of depopulation of excited electronic singlet states of molecular species, monitoring of fluorescence provides, in principle, the simplest and most direct measure of concentration. This volume comprises eight chapters, with the first focusing on the time dependence and applications of fluorescence. Succeeding chapters go on to discuss basic principles of the single photon counting lifetime measurement; light sources; photomultipliers; electronics; data analysis; nanosecond time-resolved emission spectroscopy; time dependence of fluorescence anisotropy. This book will be of interest to practitioners in the field of chemistry.

Fluorescent Methods to Study Biological Membranes

Fluorescent Methods to Study Biological Membranes
  • Author : Yves Mely,Guy Duportail
  • Publisher :Unknown
  • Release Date :2012-10-10
  • Total pages :486
  • ISBN : 9783642331275
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Summary : Biological membranes play a central role in cell structure, shape and functions. However, investigating the membrane bilayer has proved to be difficult due to its highly dynamic and anisotropic structure, which generates steep gradients at the nanometer scale. Due to the decisive impact of recently developed fluorescence-based techniques, tremendous advances have been made in the last few years in our understanding of membrane characteristics and functions. In this context, the present book illustrates some of these major advances by collecting review articles written by highly respected experts. The book is organized in three parts, the first of which deals with membrane probes and model membranes. The second part describes the use of advanced quantitative and high-resolution techniques to explore the properties of biological membranes, illustrating the key progress made regarding membrane organization, dynamics and interactions. The third part is focused on the investigation of membrane proteins using the same techniques, and notably on the membrane receptors that play a central role in signaling pathways and therapeutic strategies. All chapters provide comprehensive information on membranes and their exploration for beginners in the field and advanced researchers alike.

Methods in Systems Biology

Methods in Systems Biology
  • Author : Anonim
  • Publisher :Unknown
  • Release Date :2011-09-19
  • Total pages :668
  • ISBN : 9780123851192
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Summary : Systems biology is a term used to describe a number of trends in bioscience research and a movement that draws on those trends. This volume in the Methods in Enzymology series comprehensively covers the methods in systems biology. With an international board of authors, this volume is split into sections that cover subjects such as machines for systems biology, protein production and quantification for systems biology, and enzymatic assays in systems biology research. This volume in the Methods in Enzymology series comprehensively covers the methods in systems biology With an international board of authors, this volume is split into sections that cover subjects such as machines for systems biology, protein production and quantification for systems biology, and enzymatic assays in systems biology research

The Current Trends of Optics and Photonics

The Current Trends of Optics and Photonics
  • Author : Cheng-Chung Lee
  • Publisher :Unknown
  • Release Date :2014-11-25
  • Total pages :542
  • ISBN : 9789401793926
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Summary : Optics and photonics offer new and vibrant approaches to meeting the challenges of the 21st century concerning energy conservation, education, agriculture, personal health and the environment. One of the most effective ways to address these global problems is to provide updated and reliable content on light-based technologies. Optical thin films and meta-materials, lasers, optical communications, light-emitting diodes, solar cells, liquid crystal technology, nanophotonics and biophotonics all play vital roles in enriching our lives. We hope to raise readers’ awareness of how optical technologies are now promoting sustainable development and providing reliable solutions to basic human needs. Furthermore, in order to broaden new research fields, we hope to inspire them to pursue further cutting-edge breakthroughs on the basis of the accomplishments that have already been made.

Digital Microscopy

Digital Microscopy
  • Author : Greenfield Sluder,David E. Wolf
  • Publisher :Unknown
  • Release Date :2013-08-07
  • Total pages :672
  • ISBN : 9780124078925
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Summary : The previous edition of this book marked the shift in technology from video to digital camera use with microscope use in biological science. This new edition presents some of the optical fundamentals needed to provide a quality image to the digital camera. Specifically, it covers the fundamental geometric optics of finite- and infinity-corrected microscopes, develops the concepts of physical optics and Abbe s theory of image formation, presents the principles of Kohler illumination, and finally reviews the fundamentals of fluorescence and fluorescence microscopy. The second group of chapters deals with digital and video fundamentals: how digital and video cameras work, how to coordinate cameras with microscopes, how to deal with digital data, the fundamentals of image processing, and low light level cameras. The third group of chapters address some specialized areas of microscopy that allow sophisticated measurements of events in living cells that are below the optical limits of resolution. Expands coverage to include discussion of confocal microscopy not found in the previous edition Includes "traps and pitfalls" as well as laboratory exercises to help illustrate methods

Multiphoton Microscopy

Multiphoton Microscopy
  • Author : Espen Hartveit
  • Publisher :Unknown
  • Release Date :2020-11-15
  • Total pages :364
  • ISBN : 1493997041
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Summary : This volume covers the latest techniques and strategies used in multi-photon excitation (MPE) microscopy. Chapters in this book cover the fundamentals of MPE microscopy as applied to both in vitro and in vivo experimental systems; information on how to combine MPE microscopy with targeted electrophysiological recordings, calcium imaging, and transmembrane voltage imaging; methods to investigate cellular and large-scale neural morphology; signaling in astrocytes; and ways to use MPE microscopy to study the retina. In Neuromethods series style, chapters include the kind of detail and key advice from the specialists needed to get successful results in your laboratory. Comprehensive and thorough, Multiphoton Microscopy is a valuable resource for both expert and novice researchers interested in expanding their knowledge and research in this rapidly developing field.

Introduction to Biophotonics

Introduction to Biophotonics
  • Author : Paras N. Prasad
  • Publisher :Unknown
  • Release Date :2004-01-16
  • Total pages :593
  • ISBN : 9780471465393
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Summary : Paras Prasad’s text provides a basic knowledge of a broadrange of topics so that individuals in all disciplines can rapidlyacquire the minimal necessary background for research anddevelopment in biophotonics. Introduction to Biophotonics serves asboth a textbook for education and training as well as a referencebook that aids research and development of those areas integratinglight, photonics, and biological systems. Each chapter contains atopic introduction, a review of key data, and description of futuredirections for technical innovation. Introduction to Biophotonicscovers the basic principles of Optics Optical spectroscopy Microscopy Each section also includes illustrated examples and reviewquestions to test and advance the reader’s knowledge.Sections on biosensors and chemosensors, important tools forcombating biological and chemical terrorism, will be of particularinterest to professionals in toxicology and other environmentaldisciplines. Introduction to Biophotonics proves a valuablereference for graduate students and researchers in engineering,chemistry, and the life sciences.

Fluorescence Spectroscopy and Microscopy

Fluorescence Spectroscopy and Microscopy
  • Author : Yves Engelborghs,Antonie J. W. G. Visser
  • Publisher :Unknown
  • Release Date :2014
  • Total pages :816
  • ISBN : 1627036490
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Summary : Reflecting the expanding field's need for reliable protocols, Fluorescence Spectroscopy and Microscopy: Methods and Protocols offers techniques from a worldwide team of experts on this versatile and vital subject. The topics covered fall into four broad categories: steady-state fluorescence spectroscopy, time-resolved fluorescence spectroscopy, fluorescent probe development, and the various sub-categories of fluorescence microscopy, such as fluorescence recovery after photobleaching (FRAP), live cell FRET imaging (FRETim), fluorescence lifetime imaging (FLIM), fluorescence fluctuation spectroscopy (FFS), and single-molecule fluorescence spectroscopy (smFS). Written as a part of the popular Methods in Molecular Biology series, chapters include the kind of unambiguous detail and key implementation advice that proves essential for successful results.

Imaging Living Cells

Imaging Living Cells
  • Author : Rosario Rizzuto,Cristina Fasolato
  • Publisher :Unknown
  • Release Date :2012-12-06
  • Total pages :394
  • ISBN : 9783642600036
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Summary : Intended for bench-top use, this lab manual is suitable for both scientists and graduate students, since it combines an update on the most advanced imaging procedures with detailed protocols. Examples, carefully selected from the wide repertoire of cell pyhsiology, cover such different functional aspects as distribution of multiple ions, electrical activity, exo-endocytosis, gene expression, and the cell cycle.