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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.
Ideal for cell biologists, life scientists, biomedical engineers, and clinicians, this handbook provides comprehensive treatment of the theories, techniques, and biomedical applications of nonlinear optics and microscopy.
This comprehensive handbook gives a fully updated guide to lasers and laser technologies, including the complete range of their technical applications. This third volume covers modern applications in engineering and technology, including all new and updated case studies spanning telecommunications and data storage to medicine, optical measurement, defense and security, nanomaterials processing and characterization. Key Features: • Offers a complete update of the original, bestselling work, including many brand-new chapters. • Deepens the introduction to fundamentals, from laser design and fabrication to host matrices for solid-state lasers, energy level diagrams, hosting materials, dopant energy levels, and lasers based on nonlinear effects. • Covers new laser types, including quantum cascade lasers, silicon-based lasers, titanium sapphire lasers, terahertz lasers, bismuth-doped fiber lasers, and diode-pumped alkali lasers. • Discusses the latest applications, e.g., lasers in microscopy, high-speed imaging, attosecond metrology, 3D printing, optical atomic clocks, time-resolved spectroscopy, polarization and profile measurements, pulse measurements, and laser-induced fluorescence detection. • Adds new sections on laser materials processing, laser spectroscopy, lasers in imaging, lasers in environmental sciences, and lasers in communications. This handbook is the ideal companion for scientists, engineers, and students working with lasers, including those in optics, electrical engineering, physics, chemistry, biomedicine, and other relevant areas.
Small molecules and conjugated polymers, the two main types of organic materials used for optoelectronic and photonic devices, can be used in a number of applications including organic light-emitting diodes, photovoltaic devices, photorefractive devices and waveguides. Organic materials are attractive due to their low cost, the possibility of their deposition from solution onto large-area substrates, and the ability to tailor their properties. The Handbook of organic materials for optical and (opto)electronic devices provides an overview of the properties of organic optoelectronic and nonlinear optical materials, and explains how these materials can be used across a range of applications.Parts one and two explore the materials used for organic optoelectronics and nonlinear optics, their properties, and methods of their characterization illustrated by physical studies. Part three moves on to discuss the applications of optoelectronic and nonlinear optical organic materials in devices and includes chapters on organic solar cells, electronic memory devices, and electronic chemical sensors, electro-optic devices.The Handbook of organic materials for optical and (opto)electronic devices is a technical resource for physicists, chemists, electrical engineers and materials scientists involved in research and development of organic semiconductor and nonlinear optical materials and devices. - Comprehensively examines the properties of organic optoelectronic and nonlinear optical materials - Discusses their applications in different devices including solar cells, LEDs and electronic memory devices - An essential technical resource for physicists, chemists, electrical engineers and materials scientists
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.
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.
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