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This open access book provides a comprehensive overview of the application of the newest laser and microscope/ophthalmoscope technology in the field of high resolution imaging in microscopy and ophthalmology. Starting by describing High-Resolution 3D Light Microscopy with STED and RESOLFT, the book goes on to cover retinal and anterior segment imaging and image-guided treatment and also discusses the development of adaptive optics in vision science and ophthalmology. Using an interdisciplinary approach, the reader will learn about the latest developments and most up to date technology in the field and how these translate to a medical setting. High Resolution Imaging in Microscopy and Ophthalmology – New Frontiers in Biomedical Optics has been written by leading experts in the field and offers insights on engineering, biology, and medicine, thus being a valuable addition for scientists, engineers, and clinicians with technical and medical interest who would like to understand the equipment, the applications and the medical/biological background. Lastly, this book is dedicated to the memory of Dr. Gerhard Zinser, co-founder of Heidelberg Engineering GmbH, a scientist, a husband, a brother, a colleague, and a friend.
Serving as a practical guide to the ocular imaging modalities that are currently available to eye care providers for the care of glaucoma patients, this book provides information on advances in ocular imaging and their applications in the diagnosis and management of glaucoma. Each chapter introduces the imaging modality, highlight its strengths and weaknesses for clinical care, and discuss its integration into the clinical examination and decision-making process. The chapters also provide an in-depth description of the interpretation of images from each imaging modality. When appropriate, the chapters will summarize past and ongoing research and propose future research directions and clinical applications. This title will appeal to ophthalmologists and optometrists at all levels, from trainees to experienced clinicians looking to learn new and important information.
This book offers an overview of imaging techniques used to investigate cells and tissue in their native environment. It covers the range of imaging approaches used, as well as the application of those techniques to the study of biological processes in cells and whole tissues within living organisms.
The Encyclopedia of Modern Optics, Second Edition, Five Volume Set provides a wide-ranging overview of the field, comprising authoritative reference articles for undergraduate and postgraduate students and those researching outside their area of expertise. Topics covered include classical and quantum optics, lasers, optical fibers and optical fiber systems, optical materials and light-emitting diodes (LEDs). Articles cover all subfields of optical physics and engineering, such as electro-optical design of modulators and detectors. This update contains contributions from international experts who discuss topics such as nano-photonics and plasmonics, optical interconnects, photonic crystals and 2D materials, such as graphene or holy fibers. Other topics of note include solar energy, high efficiency LED’s and their use in illumination, orbital angular momentum, quantum optics and information, metamaterials and transformation optics, high power fiber and UV fiber lasers, random lasers and bio-imaging. Addresses recent developments in the field and integrates concepts from fundamental physics with applications for manufacturing and engineering/design Provides a broad and interdisciplinary coverage of specialist areas Ensures that the material is appropriate for new researchers and those working in a new sub-field, as well as those in industry Thematically arranged and alphabetically indexed, with cross-references added to facilitate ease-of-use
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.
This book focuses on the emerging non-invasive imaging technique of Fluorescence Lifetime Imaging Ophthalmoscopy (FLIO). FLIO reveals unique information on retinal diseases, ranging from age-related macular degeneration and vascular diseases to hereditary retinal dystrophies. Fluorescence lifetimes enable the evaluation of disease progression before irreversible structural changes occur. The image acquisition is suitable for diagnostic purposes and follow-up examinations to investigate the natural course of disease, and to monitor the effects of possible therapies. This book fills the gap between available literature and gives state-of-the-art guidance on the principles of the FLIO technique, image acquisition, and data analysis. Written by a team of expert leaders within this field, this book will be relevant for scientists and clinicians with an interest in ophthalmoscopy.
Quantitative bioimaging is a broad interdisciplinary field that exploits tools from biology, chemistry, optics, and statistical data analysis for the design and implementation of investigations of biological processes. Instead of adopting the traditional approach of focusing on just one of the component disciplines, this textbook provides a unique introduction to quantitative bioimaging that presents all of the disciplines in an integrated manner. The wide range of topics covered include basic concepts in molecular and cellular biology, relevant aspects of antibody technology, instrumentation and experimental design in fluorescence microscopy, introductory geometrical optics and diffraction theory, and parameter estimation and information theory for the analysis of stochastic data. Key Features: Comprises four parts, the first of which provides an overview of the topics that are developed from fundamental principles to more advanced levels in the other parts. Presents in the second part an in-depth introduction to the relevant background in molecular and cellular biology and in physical chemistry, which should be particularly useful for students without a formal background in these subjects. Provides in the third part a detailed treatment of microscopy techniques and optics, again starting from basic principles. Introduces in the fourth part modern statistical approaches to the determination of parameters of interest from microscopy data, in particular data generated by single molecule microscopy experiments. Uses two topics related to protein trafficking (transferrin trafficking and FcRn-mediated antibody trafficking) throughout the text to motivate and illustrate microscopy techniques. An online appendix providing the background and derivations for various mathematical results presented or used in the text is available at http://www.routledge.com/9781138598980.
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.