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Advanced Time Correlated Single Photon Counting Applications


Advanced Time Correlated Single Photon Counting Applications
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Advanced Time Correlated Single Photon Counting Techniques


Advanced Time Correlated Single Photon Counting Techniques
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Author : Wolfgang Becker
language : en
Publisher: Springer Science & Business Media
Release Date : 2005-12-19

Advanced Time Correlated Single Photon Counting Techniques written by Wolfgang Becker and has been published by Springer Science & Business Media this book supported file pdf, txt, epub, kindle and other format this book has been release on 2005-12-19 with Science categories.


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.



Advanced Time Correlated Single Photon Counting Applications


Advanced Time Correlated Single Photon Counting Applications
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Author : Wolfgang Becker
language : en
Publisher: Springer
Release Date : 2015-04-13

Advanced Time Correlated Single Photon Counting Applications written by Wolfgang Becker and has been published by Springer this book supported file pdf, txt, epub, kindle and other format this book has been release on 2015-04-13 with Science categories.


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.



Time Correlated Single Photon Counting


Time Correlated Single Photon Counting
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Author : Desmond O'Connor
language : en
Publisher: Academic Press
Release Date : 2012-12-02

Time Correlated Single Photon Counting written by Desmond O'Connor and has been published by Academic Press this book supported file pdf, txt, epub, kindle and other format this book has been release on 2012-12-02 with Science categories.


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.



Advanced Photon Counting


Advanced Photon Counting
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Author : Peter Kapusta
language : en
Publisher: Springer
Release Date : 2015-04-23

Advanced Photon Counting written by Peter Kapusta and has been published by Springer this book supported file pdf, txt, epub, kindle and other format this book has been release on 2015-04-23 with Science categories.


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.



Advanced Optical Methods For Brain Imaging


Advanced Optical Methods For Brain Imaging
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Author : Fu-Jen Kao
language : en
Publisher: Springer
Release Date : 2018-05-14

Advanced Optical Methods For Brain Imaging written by Fu-Jen Kao and has been published by Springer this book supported file pdf, txt, epub, kindle and other format this book has been release on 2018-05-14 with Technology & Engineering categories.


This book highlights the rapidly developing field of advanced optical methods for structural and functional brain imaging. As is known, the brain is the most poorly understood organ of a living body. It is indeed the most complex structure in the known universe and, thus, mapping of the brain has become one of the most exciting frontlines of contemporary research. Starting from the fundamentals of the brain, neurons and synapses, this book presents a streamlined and focused coverage of the core principles, theoretical and experimental approaches, and state-of-the-art applications of most of the currently used imaging methods in brain research. It presents contributions from international leaders on different photonics-based brain imaging modalities and techniques. Included are comprehensive descriptions of many of the technology driven spectacular advances made over the past few years that have allowed novel insights of the structural and functional details of neurons. The book is targeted at researchers, engineers and scientists who are working in the field of brain imaging, neuroscience and connectomics. Although this book is not intended to serve as a textbook, it will appeal to undergraduate students engaged in the specialization of brain imaging.



The Bh Tcspc Handbook


The Bh Tcspc Handbook
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Author : Dr. Wolfgang Becker
language : en
Publisher: Becker & Hickl GmbH
Release Date : 2021-09-01

The Bh Tcspc Handbook written by Dr. Wolfgang Becker and has been published by Becker & Hickl GmbH this book supported file pdf, txt, epub, kindle and other format this book has been release on 2021-09-01 with Science categories.


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.



Multiphoton Microscopy And Fluorescence Lifetime Imaging


Multiphoton Microscopy And Fluorescence Lifetime Imaging
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Author : Karsten König
language : en
Publisher: Walter de Gruyter GmbH & Co KG
Release Date : 2018-01-22

Multiphoton Microscopy And Fluorescence Lifetime Imaging written by Karsten König and has been published by Walter de Gruyter GmbH & Co KG this book supported file pdf, txt, epub, kindle and other format this book has been release on 2018-01-22 with Science categories.


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



Modern Tools For Time Resolved Luminescence Biosensing And Imaging


Modern Tools For Time Resolved Luminescence Biosensing And Imaging
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Author :
language : en
Publisher: Frontiers Media SA
Release Date : 2021-09-01

Modern Tools For Time Resolved Luminescence Biosensing And Imaging written by and has been published by Frontiers Media SA this book supported file pdf, txt, epub, kindle and other format this book has been release on 2021-09-01 with Science categories.




Fluorescence Spectroscopy And Microscopy In Biology


Fluorescence Spectroscopy And Microscopy In Biology
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Author : Radek Šachl
language : en
Publisher: Springer Nature
Release Date : 2023-04-27

Fluorescence Spectroscopy And Microscopy In Biology written by Radek Šachl and has been published by Springer Nature this book supported file pdf, txt, epub, kindle and other format this book has been release on 2023-04-27 with Science categories.


This book provides the reader with an updated comprehensive view of the rapidly developing and fascinating field of fluorescence spectroscopy and microscopy. In recent years, fluorescence spectroscopy and microscopy have experienced rapid technological development, which has enabled the detection and monitoring of single molecules with high spatial and temporal resolution. Thanks to these developments, fluorescence has become an even more popular method in physical, biological and related fields. This book guides the reader through both basic and advanced fluorescence spectroscopy and microscopy approaches with a focus on their applications in membrane and protein biophysics. Each of the four parts: A - Fluorescence Spectroscopy, B - Fluorescence Microscopy, C - Applications of Fluorescence Spectroscopy and Microscopy to biological membranes and D - Applications of Fluorescence Spectroscopy to protein studies are written by experts within the field. The book is intended for both complete beginners who want to quickly orient themselves in the large number of existing fluorescent methods, as well as for advanced readers who are interested in particular methods and their proper use.



Multi Parametric Live Cell Microscopy Of 3d Tissue Models


Multi Parametric Live Cell Microscopy Of 3d Tissue Models
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Author : Ruslan I. Dmitriev
language : en
Publisher: Springer
Release Date : 2017-10-26

Multi Parametric Live Cell Microscopy Of 3d Tissue Models written by Ruslan I. Dmitriev and has been published by Springer this book supported file pdf, txt, epub, kindle and other format this book has been release on 2017-10-26 with Science categories.


This book provides an essential overview of existing state-of-the-art quantitative imaging methodologies and protocols (intensity-based ratiometric and FLIM/ PLIM). A variety of applications are covered, including multi-parametric quantitative imaging in intestinal organoid culture, autofluorescence imaging in cancer and stem cell biology, Ca2+ imaging in neural ex vivo tissue models, as well as multi-parametric imaging of pH and viscosity in cancer biology. The current state-of-the-art of 3D tissue models and their compatibility with live cell imaging is also covered. This is an ideal book for specialists working in tissue engineering and designing novel biomaterial.