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FRET and FLIM Techniques

FRET and FLIM Techniques Book
Author : Theodorus W. J. Gadella
Publisher : Elsevier
Release : 2011-07-29
ISBN : 9780080915128
Language : En, Es, Fr & De

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Book Description :

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.

FRET and FLIM Techniques

FRET and FLIM Techniques Book
Author : Theodorus W. J. Gadella
Publisher : Elsevier Science
Release : 2009-01-05
ISBN : 9780080549583
Language : En, Es, Fr & De

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Book Description :

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 Book
Author : Ammasi Periasamy,Richard Day
Publisher : Elsevier
Release : 2011-04-28
ISBN : 9780080536873
Language : En, Es, Fr & De

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Book Description :

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.

Fluorescence Spectroscopy and Microscopy

Fluorescence Spectroscopy and Microscopy Book
Author : Yves Engelborghs,Antonie J.W.G. Visser
Publisher : Humana Press
Release : 2016-08-23
ISBN : 9781493963157
Language : En, Es, Fr & De

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Book Description :

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. Comprehensive and practical, Fluorescence Spectroscopy and Microscopy: Methods and Protocols aims to guide both ‘novice’ and established scientists toward furthering their research with these invaluable techniques.

FLIM Microscopy in Biology and Medicine

FLIM Microscopy in Biology and Medicine Book
Author : Ammasi Periasamy,Robert M. Clegg
Publisher : CRC Press
Release : 2009-07-06
ISBN : 9781420078916
Language : En, Es, Fr & De

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Book Description :

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.

FRET and FLIM Techniques

FRET and FLIM Techniques Book
Author : Theodorus W. J. Gadella
Publisher : Elsevier Science
Release : 2009-01-05
ISBN : 9780080549583
Language : En, Es, Fr & De

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Book Description :

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.

Methods in Cellular Imaging

Methods in Cellular Imaging Book
Author : Ammasi Periasamy
Publisher : Springer
Release : 2013-05-27
ISBN : 1461475139
Language : En, Es, Fr & De

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Book Description :

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.

Methods and Techniques in Drug Discovery

Methods and Techniques in Drug Discovery Book
Author : John Sterling,Ellyn J. T. D. Kerr,Shannon Simons
Publisher : Unknown
Release : 2005
ISBN : 0987650XXX
Language : En, Es, Fr & De

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Book Description :

Download Methods and Techniques in Drug Discovery book written by John Sterling,Ellyn J. T. D. Kerr,Shannon Simons, available in PDF, EPUB, and Kindle, or read full book online anywhere and anytime. Compatible with any devices.

Studies of the MTOR Signalling Pathway Using Advanced FRET FLIM Technique

Studies of the MTOR Signalling Pathway Using Advanced FRET FLIM Technique Book
Author : Rahul B. Yadav
Publisher : Unknown
Release : 2010
ISBN : 0987650XXX
Language : En, Es, Fr & De

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Book Description :

Download Studies of the MTOR Signalling Pathway Using Advanced FRET FLIM Technique book written by Rahul B. Yadav, available in PDF, EPUB, and Kindle, or read full book online anywhere and anytime. Compatible with any devices.

Advanced Photon Counting

Advanced Photon Counting Book
Author : Peter Kapusta,Michael Wahl,Rainer Erdmann
Publisher : Springer
Release : 2015-04-23
ISBN : 3319156365
Language : En, Es, Fr & De

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Book Description :

This volume focuses on Time-Correlated Single Photon Counting (TCSPC), a powerful tool allowing luminescence lifetime measurements to be made with high temporal resolution, even on single molecules. Combining spectrum and lifetime provides a “fingerprint” for identifying such molecules in the presence of a background. Used together with confocal detection, this permits single-molecule spectroscopy and microscopy in addition to ensemble measurements, opening up an enormous range of hot life science applications such as fluorescence lifetime imaging (FLIM) and measurement of Förster Resonant Energy Transfer (FRET) for the investigation of protein folding and interaction. Several technology-related chapters present both the basics and current state-of-the-art, in particular of TCSPC electronics, photon detectors and lasers. The remaining chapters cover a broad range of applications and methodologies for experiments and data analysis, including the life sciences, defect centers in diamonds, super-resolution microscopy, and optical tomography. The chapters detailing new options arising from the combination of classic TCSPC and fluorescence lifetime with methods based on intensity fluctuation represent a particularly unique highlight.

Fluorescence Lifetime Spectroscopy and Imaging

Fluorescence Lifetime Spectroscopy and Imaging Book
Author : Laura Marcu,Paul M. W. French,Daniel S. Elson
Publisher : CRC Press
Release : 2014-07-17
ISBN : 1439861676
Language : En, Es, Fr & De

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Book Description :

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 Diagnostics explores the remarkable advances in time-resolved fluorescence techniques and their role in a wide range of biological and clinical applications. Broadly accessible, the book captures the state-of-the-art of fluorescence lifetime metrology and imaging and provides current perspectives on their applications to biomedical studies of intact tissues and medical diagnosis. The text introduces these techniques within the wider context of fluorescence spectroscopy and describes basic principles underlying current instrumentation for fluorescence lifetime imaging and metrology (FLIM). It also covers the wide range of methods, including single channel (point) spectroscopy, fluorescence lifetime imaging microscopy, and single- and multi-photon excitation. Edited by pioneers in this field, with contributions from leading experts, the book includes an overview of complementary techniques that help researchers beginning FLIM research. It offers a comprehensive treatment of fundamental principles, instrumentation, analytical methods, and applications. It also provides an overview of the label-free contrast available from lifetime measurements of tissue autofluorescence and the prospects for exploiting this for clinical applications and biomedical research including drug discovery.

Advanced Time Correlated Single Photon Counting Applications

Advanced Time Correlated Single Photon Counting Applications Book
Author : Wolfgang Becker
Publisher : Springer
Release : 2015-04-13
ISBN : 3319149296
Language : En, Es, Fr & De

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Book Description :

This book is an attempt to bridge the gap between the instrumental principles of multi-dimensional time-correlated single photon counting (TCSPC) and typical applications of the technique. Written by an originator of the technique and by sucessful users, it covers the basic principles of the technique, its interaction with optical imaging methods and its application to a wide range of experimental tasks in life sciences and clinical research. The book is recommended for all users of time-resolved detection techniques in biology, bio-chemistry, spectroscopy of live systems, live cell microscopy, clinical imaging, spectroscopy of single molecules, and other applications that require the detection of low-level light signals at single-photon sensitivity and picosecond time resolution.

Understanding Light Microscopy

Understanding Light Microscopy Book
Author : Jeremy Sanderson
Publisher : Wiley
Release : 2019-05-28
ISBN : 0470973757
Language : En, Es, Fr & De

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Book Description :

Introduces readers to the enlightening world of the modern light microscope There have been rapid advances in science and technology over the last decade, and the light microscope, together with the information that it gives about the image, has changed too. Yet the fundamental principles of setting up and using a microscope rests upon unchanging physical principles that have been understood for years. This informative, practical, full-colour guide fills the gap between specialised edited texts on detailed research topics, and introductory books, which concentrate on an optical approach to the light microscope. It also provides comprehensive coverage of confocal microscopy, which has revolutionised light microscopy over the last few decades. Written to help the reader understand, set up, and use the often very expensive and complex modern research light microscope properly, Understanding Light Microscopy keeps mathematical formulae to a minimum—containing and explaining them within boxes in the text. Chapters provide in-depth coverage of basic microscope optics and design; ergonomics; illumination; diffraction and image formation; reflected-light, polarised-light, and fluorescence microscopy; deconvolution; TIRF microscopy; FRAP & FRET; super-resolution techniques; biological and materials specimen preparation; and more. Gives a didactic introduction to the light microscope Encourages readers to use advanced fluorescence and confocal microscopes within a research institute or core microscopy facility Features full-colour illustrations and workable practical protocols Understanding Light Microscopy is intended for any scientist who wishes to understand and use a modern light microscope. It is also ideal as supporting material for a formal taught course, or for individual students to learn the key aspects of light microscopy through their own study.

Microscopy Techniques

Microscopy Techniques Book
Author : Jens Rietdorf
Publisher : Springer Science & Business Media
Release : 2005-06-23
ISBN : 9783540236986
Language : En, Es, Fr & De

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Book Description :

With contributions by numerous experts

FLIM Microscopy in Biology and Medicine

FLIM Microscopy in Biology and Medicine Book
Author : Ammasi Periasamy,Robert M. Clegg
Publisher : Chapman and Hall/CRC
Release : 2010
ISBN : 0987650XXX
Language : En, Es, Fr & De

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Book Description :

After an overview of basic principles of fluorescence, microscopy, and fluorescence lifetime imaging (FLIM), this book discusses probes, dynamic and video rate FLIM imaging, laser scanning confocal FLIM, spectral lifetime imaging, nonlinear curve-fitting methods, global FLIM data analysis, and new approaches in data analysis.

Multiphoton Microscopy and Fluorescence Lifetime Imaging

Multiphoton Microscopy and Fluorescence Lifetime Imaging Book
Author : Karsten König
Publisher : Walter de Gruyter GmbH & Co KG
Release : 2018-01-22
ISBN : 3110429985
Language : En, Es, Fr & De

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Book Description :

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

Measuring Force in the Developing Zebrafish Embryo Using an Expressible Tension Sensor

Measuring Force in the Developing Zebrafish Embryo Using an Expressible Tension Sensor Book
Author : Andrea Leigh Hamilton
Publisher : Unknown
Release : 2015
ISBN : 0987650XXX
Language : En, Es, Fr & De

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Book Description :

Early vertebrate development is a mechanically dynamic process. Embryos undergo radical morphologic changes to mold a ball of cells into the recognizable planes of a frog or fish or mouse. In the last 40 years, the zebrafish has emerged as a powerful system for studying these early movements in a vertebrate system. Its large, optically transparent embryos make it a good candidate for advanced microscopy techniques, and in recent years this model organism has been the setting for a host of cutting edge microscopy methodologies. One technique that has been comparatively overlooked in the fish is the use of Fluorescence Lifetime Imaging (FLIM), specifically in conjunction with Forster Resonance Energy Transfer (FRET). FRET is a powerful technique for studying real time, in vivo protein dynamics on the nanometer scale. FLIM allows us to apply this technique in the noisy embryo background without the need for cumbersome spectral bleed-through corrections. In this work, we adapt previously published controls for use in the zebrafish system and then take a systematic approach to developing the best analytical practices for FLIM-FRET in the zebrafish embryo. After developing FLIM-FRET in the early embryo background, we adapt a recently published expressible, FRET-based stress sensor capable of measuring piconewton levels of force in vivo (Grashoff et. al. 2010). We insert this Tension Sensor Module (TSMod) into the zebrafish Epithelial Cell Adhesion Molecule (EpCAM) and use it to make direct measurements of the tensions experienced during early development. In doing so we show that the EpCAM molecule holds between 0.5 and 1.5 piconewtons of force during zebrafish epiboly and that the leading margin is under greater tension than other regions of the developing embryo. This work represents the first use of FLIM-FRET in the zebrafish embryo and the first direct measure of molecular tension in a developing vertebrate.

Transactions

Transactions Book
Author : Biochemical Society (Great Britain)
Publisher : Unknown
Release : 2006
ISBN : 0987650XXX
Language : En, Es, Fr & De

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Book Description :

Download Transactions book written by Biochemical Society (Great Britain), available in PDF, EPUB, and Kindle, or read full book online anywhere and anytime. Compatible with any devices.

Chemtracts

Chemtracts Book
Author : Anonim
Publisher : Unknown
Release : 2008
ISBN : 0987650XXX
Language : En, Es, Fr & De

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Book Description :

Consists of reviews, condensations, and commentaries.