FLIM Microscopy in Biology and Medicine PDF Download

Author: Ammasi Periasamy
Publisher: CRC Press
ISBN: 1420078917
Category : Science
Languages : en
Pages : 474

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Book Description
Detecting Signals at the Single Molecule Level: Pioneering Achievements in MicroscopyRecent 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 be

Author: Karsten König
Publisher: Walter de Gruyter GmbH & Co KG
ISBN: 311043007X
Category : Science
Languages : en
Pages : 785

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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

Author: Ammasi Periasamy
Publisher: Chapman and Hall/CRC
ISBN:
Category : Medical
Languages : en
Pages : 480

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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.

Author: Theodorus W. J. Gadella
Publisher: Elsevier
ISBN: 0080915124
Category : Science
Languages : en
Pages : 603

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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.

Author: Desmond O'Connor
Publisher: Academic Press
ISBN: 0323141447
Category : Science
Languages : en
Pages : 299

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Book Description
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.

Author: Ammasi Periasamy
Publisher: Springer
ISBN: 1461475139
Category : Science
Languages : en
Pages : 473

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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.

Author: Rongguang Liang
Publisher: Springer Science & Business Media
ISBN: 364228390X
Category : Science
Languages : en
Pages : 391

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Book Description
This book provides an introduction to design of biomedical optical imaging technologies and their applications. The main topics include: fluorescence imaging, confocal imaging, micro-endoscope, polarization imaging, hyperspectral imaging, OCT imaging, multimodal imaging and spectroscopic systems. Each chapter is written by the world leaders of the respective fields, and will cover: principles and limitations of optical imaging technology, system design and practical implementation for one or two specific applications, including design guidelines, system configuration, optical design, component requirements and selection, system optimization and design examples, recent advances and applications in biomedical researches and clinical imaging. This book serves as a reference for students and researchers in optics and biomedical engineering.

Author: Peng Xi
Publisher: CRC Press
ISBN: 1466586303
Category : Science
Languages : en
Pages : 268

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Book Description
Microscopy is at the forefront of multidisciplinary research. It was developed by physicists, made specific by chemists, and applied by biologists and doctors to better understand how the human body works. For this very reason, the field has been revolutionized in past decades.The objective of Optical Nanoscopy and Novel Microscopy Techniques is to

Author: Dr. Wolfgang Becker
Publisher: Becker & Hickl GmbH
ISBN:
Category : Science
Languages : en
Pages : 995

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Book Description
Time-Correlated Single Photon Counting Modules SPC-130EMN, SPC-130EMNX, SPC-130IN, SPC-130INX, SPC-150N, SPC-150NX, SPC-150NXX, SPC-160, SPC-160PCIE, SPC-180N, SPC-180NX, SPC-180NXX Detectors, Lasers and Peripheral Devices Simple-Tau Systems Technical Principles TCSPC Applications FLIM Systems Applications in Life Sciences Clinical FLIM Applications SPCM Software SPCImage NG Data Analysis Software Time-correlated single photon counting (TCSPC) is an amazingly sensitive technique for recording low-level light signals with picosecond resolution and extremely high precision.TCSPC originates from the measurement of excited nuclear states and has been used since the late 60s [775, 1250]. For many years TCSPC was used primarily to record fluorescence decay curves of organic dyes in solution. Due to the low intensity and low repetition rate of the light sources and the limited speed of the electronics of the 70s and 80s the acquisition times were extremely long. More important, classic TCSPC was intrinsically one-dimensional, i.e. limited to the recording of the waveform of a periodic light signal. Light sources ceased to be a limitation when the first mode-locked Argon lasers and synchronously pumped dye lasers were introduced. For the recording electronics, the situation changed with the introduction of the SPC-300 modules of Becker & Hickl in 1993. Due to a new analog-to-digital conversion principle these modules could be used at photon count rates almost 100 times higher than the classic TCSPC devices. Moreover, the modules were able to record the photons of a large number of detectors simultaneously. They were thus able to record a photon distribution not only versus the time in a fluorescence decay but also versus aspatial coordinate or the wavelength of the photons. Multi-dimensional TCSPC was born. Within a few years, more dimensions were added to multidimensional TCSPC. Fast sequential recording was introduced with the SPC-430 in 1995, fast scanning with the SPC-535 in 1997. Time-tag recording was introduced with the SPC-431 in 1996; multi-module TCSPC systems followed in 1999. Since then, the Becker & Hickl TCSPC systems became bigger, faster and more flexible. Recent TCSPC modules, like the SPC-150NX or the SPC-180, can be configured for sequential recording, imaging, or time-tag recording by a simple software command. Multi-module systems, like the SPC-134EM and SPC-154, can be used for scanning at unprecedented count rates and acquisition speeds. Nevertheless, TCSPC still has the reputation to be an extremely sluggish technique unable to record any fast changes in the fluorescence or scattering behaviour of a sample. The multidimensional features of modern TCSPC are not commonly understood. Thus, many users do not make efficient use of their SPC modules. However, if appropriately used, multidimensional TCSPC techniques not only deliver superior results but also solve highly sophisticated measurement problems. This handbook is an attempt to help existing and potential users understand and make use of the advanced features of modern TCSPC. After an introduction into the bh TCSPC devices and associated detector, laser, and experiment control modules the principles of advanced TCSPC techniques are described. These include multidetector TCSPC, multiplexed TCSPC, sequential recording techniques, scanning techniques, parameter-tag recording, and multi-module TCSPC techniques. The next chapter describes the architecture of the bh SPC modules. A chapter about detectors gives a review of detector principles and of the parameters used to characterise detectors. It describes a number of detectors commonly used for TCSPC and gives advice about obtaining best performance from them. The implementation of bh SPC devices is described in the next part of the handbook. It includes principles and wiring diagrams for typical experiments, guidelines for first system setup, and advice for system optimisation. It describes dead-time, counting loss, and pile-up effects, detector effects, and effects related to the optical system. The next chapter of the handbook is dedicated to TCSPC applications. The first part of this chapter describes the measurement of fluorescence and anisotropy decay curves, multispectral lifetime experiments, recording of transient fluorescence lifetime phenomena, and measurements of phosphorescence decay curves. The second part of the chapter is dedicated to time-resolved laser scanning microscopy. It contains sections on a wide variety of fluorescence-lifetime imaging (FLIM) experiments and procedures, such as FLIM with various excitation principles, excitation sources, and detection principles, high-speed and time-series FLIM, Z-stack FLIM, simultaneous fluorescence and phosphorescence lifetime imaging (FLIM/PLIM), fluorescence lifetime-transient scanning (FLITS), and FLIM with special microscope configurations. A third part contains FLIM background knowledge: Signal-to-noise ratio, acquisition time, the effect of counting loss and pile-up, photobleaching, and fluorescence depolarisation on the recorded data. The book contains a large chapter on TCSPC applications, most of them in Biology. It contains sections on FLIM of molecular environment parameters in tissue, FLIM-based FRET measurements in cells, autofluorescence FLIM of biological tissue, plant physiology, and clinical FLIM applications. A section about diffuse optical tomography (DOT) by NIRS techniques includes breast imaging, static and functional brain imaging, perfusion measurement in the human brain, diffuse tissue spectroscopy, and small-animal imaging. Picosecond photon correlation, fluorescence correlation spectroscopy, burst-integrated fluorescence lifetime techniques, and photon counting histogram techniques are reviewed in the next sections. The last part of the application chapter gives an review of non-biological TCSPC applications like positron lifetime measurement, measurement of barrier discharges, remote sensing, metrological applications, and characterisation of detectors. The application chapter also includes practical hints about optical systems, detectors, and other technical aspects of the applications described. Another large chapter describes the SPCM operating software of the bh SPC modules. It describes the various user interface configurations, operation modes, the system and control parameters, the handling and display of the multidimensional data recorded by the modules, and the associated data file structure. The TCSPC Handbook also contains a chapter on the SPCImage NG fluorescence decay and FLIM data analysis software. It describes the general principles of fluorescence decay analysis, the calculation of fluorescence decay parameters and lifetime images by various decay models, pseudo-global analysis, multi-wavelength FLIM analysis, batch-processing of FLIM series, and analysis of PLIM data. The handbook ends with a list of more than 1200 references related to TCSPC, most of them being applications of the bh SPC devices.

Author: Ammasi Periasamy
Publisher: Elsevier
ISBN: 0080536875
Category : Science
Languages : en
Pages : 329

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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.