Indocyanine green (ICG) fluorescence has been used for imaging purposes for more than half a century; First employed by ophthalmologists for visualizing the retinal artery in the late 1960s, the application of ICG fluorescence imaging has since been continuously expanded. Recently, advances in imaging technologies have led to renewed attention regarding the use of ICG in the field of hepatobiliary surgery, as a new tool for visualizing the biliary tree and liver tumors.
This book reviews the most recent developments of fluorescent imaging techniques for medicinal chemistry research and biomedical applications, including cell imaging, in vitro diagnosis and in vivo imaging. Fluorescent imaging techniques play an important role in basic research, drug discovery and clinical translation. They have great impact to many fields including chemical biology, cell biology, medical imaging, cancer diagnosis and treatment, pharmaceutical science, among others, and they have facilitated our understanding of diseases and helped to develop many novel powerful tools for imaging and treatment of diseases. This book will appeal to scientists from numerous fields such as chemistry, pharmaceutical science, biology, materials science, and medicine, and it will serve as a very useful and handy resource for readers with different levels of scientific knowledge, ranging from entry level to professional level.
This thesis advances the long-standing challenge of measuring oxidative stress and deciphering its underlying mechanisms, and also outlines the advantages and limitations of existing design strategies. It presents a range of approaches for the chemical synthesis of fluorescent probes that detect reversible changes in cellular oxidative stress. The ability to visualise cellular processes in real-time is crucial to understanding disease development and streamline treatment, and this can be achieved using fluorescent tools that can sense reversible disturbances in cellular environments during pathogenesis. The perturbations in cellular redox state are of particular current interest in medical research, since oxidative stress is implicated in the pathogenesis of a number of diseases. The book investigates different strategies used to achieve ratiometric fluorescence output of the reversible redox probes, which nullify concentration effects associated with intensity-based probes. It also describes suitable approaches to target these probes to specific cellular organelles, thereby enabling medical researchers to visualise sub-cellular oxidative stress levels, and addressing the typically poor uptake of chemical tools into biological studies. In total it reports on four new probes that are now being used by over twenty research groups around the globe, and two of which have been commercialised. The final chapters of this thesis demonstrate successful applications of the sensors in a variety of biological systems ranging from prokaryotes to mammalian cells and whole organisms. The results described clearly indicate the immense value of collaborative, cross-disciplinary research.
To describe principles of optical imaging including chemistry and physics of fluorescence, limitations/advantages of optical imaging compared to metabolic and anatomic imaging. Describe hardware adapted for small animal imaging and for clinical applications: endoscopes and operative microscopes. Outline FDA approved and newer optical imaging probes. Include discussion of chemistry and linkage to other proteins. Review current techniques to image cancer and the development of techniques to specifically image cancer cells. Review use of exploiting differences in tissue autofluorescence to diagnose and treat cancer. Include agents such as 5-aminoleculinic acid. Review mechanisms that require proteolytic processing within the tumor to become active fluorophores. Review use of cancer selective proteins to localize probes to cancer cells: include toxins, antibodies, and minibodies. Introduction of plasmids, viruses or other genetic material may be used to express fluorescent agents in vivo. This chapter will review multiple vectors and delivery mechanisms of optical imaging cassettes.Preclinical investigations into the use of optical contrast agents for the detection of primary tumors in conventional and orthotopic models will be discussed. Preclinical investigations into the use of optical contrast agents for the detection of metastatic tumors in mouse models will be discussed. Use of targeted and non-specific optical contrast agents have been used for the detection of sentinel lymph node detection. These applications and how they differ from other applications will be discussed. Because of the unique difficulty of identifying tumor from normal tissue in brain tissue, a separate chapter would be needed. More clinical data is available for this cancer type than any other. Discussion of potential clinical applications for optical imaging and an assessment of the potential market.
Abstract: We have proposed a method for hyper-spectral fluorescent imaging based on acousto-optical filtering. The object of interest was pumped using ultraviolet radiation of mercury lamp equipped with monochromatic excitation filter with the window of transparency centered at 365 nm. Double TeO2 -based acousto-optical filter, tunable in range from 430 to 780 nm and having 2 nm bandwidth of spectral transparency, was used in order to detect quasimonochromatic images of object fluorescence. Modulating of ultraviolet pump intensity was used in order to reduce an impact of non-fluorescent background on the sample fluorescent imaging. The technique for signal-to-noise ratio improvement, based on fluorescence intensity estimation via digital processing of modulated video sequence of fluorescent object, was introduced. We have implemented the proposed technique for the test sample studying and we have discussed its possible applications.
Fluorescence Microscopy: Super-Resolution and other Novel Techniques delivers a comprehensive review of current advances in fluorescence microscopy methods as applied to biological and biomedical science. With contributions selected for clarity, utility, and reproducibility, the work provides practical tools for investigating these ground-breaking developments. Emphasizing super-resolution techniques, light sheet microscopy, sample preparation, new labels, and analysis techniques, this work keeps pace with the innovative technical advances that are increasingly vital to biological and biomedical researchers. With its extensive graphics, inter-method comparisons, and tricks and approaches not revealed in primary publications, Fluorescence Microscopy encourages readers to both understand these methods, and to adapt them to other systems. It also offers instruction on the best visualization to derive quantitative information about cell biological structure and function, delivering crucial guidance on best practices in related laboratory research. Presents a timely and comprehensive review of novel techniques in fluorescence imaging as applied to biological and biomedical research Offers insight into common challenges in implementing techniques, as well as effective solutions
This text provides a state of the art overview of tools for guiding surgeons in the modern operating room. The text explains how many modalities in the current armamentarium of radiologic imaging have been brought to the operating room for real time use. It also explains the current use of near infrared, fluorescent, and chemo-luminescent imaging to guide minimally invasive and open surgery to improve outcome. The book is separated into two sections. The first, discusses the biologic principles that underlie novel visualization of normal organs and pathology. The currently available equipment and equipment anticipated in the near future is covered. The second section summarizes current clinical applications of advanced imaging and visualization in the OR. Novel means of visualizing normal anatomic structures such as nerves, bile duct, and vessels that enhance safety of many operations are covered. Novel biologic imaging using radio-labeled and fluorescent-labeled molecular probes that allow identification of inflammation, vascular abnormalities, and cancer are also discussed. Authored by scientists who pioneer research in optics and radiology, tool makers who use this knowledge to make surgical equipment, and surgeons who innovate the field of surgery using these new operative tools, Imaging and Visualization in the Modern Operating Room is a valuable guide for surgeons, residents and fellows entering the field.
There is an ever-increasing number of genes that have been sequenced but are of completely unknown function. The ability to determine the location of such gene products within the cell, either by the use of antibodies or by the production of chimeras with green fluorescent protein, is a vital step towards understanding what they do. This is one major reason why fluorescence microscopy is enjoying a revival. This no-nonsense guide provides detailed, practical advice on all aspects of the subject: from choosing the right equipment, to interpreting results. It balances the advantages of a wide range of techniques - including live cell work - against the potential pitfalls, offering invaluable "tricks of the trade" along the way. Protein Localization by Fluorescence Light Microscopy: A Practical Approach has something to offer all microscopists, giving a solid grounding to the novice whilst extending the range of the experienced user.
Comprehensive Biomaterials brings together the myriad facets of biomaterials into one, major series of six edited volumes that would cover the field of biomaterials in a major, extensive fashion: Volume 1: Metallic, Ceramic and Polymeric Biomaterials Volume 2: Biologically Inspired and Biomolecular Materials Volume 3: Methods of Analysis Volume 4: Biocompatibility, Surface Engineering, and Delivery Of Drugs, Genes and Other Molecules Volume 5: Tissue and Organ Engineering Volume 6: Biomaterials and Clinical Use Experts from around the world in hundreds of related biomaterials areas have contributed to this publication, resulting in a continuum of rich information appropriate for many audiences. The work addresses the current status of nearly all biomaterials in the field, their strengths and weaknesses, their future prospects, appropriate analytical methods and testing, device applications and performance, emerging candidate materials as competitors and disruptive technologies, and strategic insights for those entering and operational in diverse biomaterials applications, research and development, regulatory management, and commercial aspects. From the outset, the goal was to review materials in the context of medical devices and tissue properties, biocompatibility and surface analysis, tissue engineering and controlled release. It was also the intent both, to focus on material properties from the perspectives of therapeutic and diagnostic use, and to address questions relevant to state-of-the-art research endeavors. Reviews the current status of nearly all biomaterials in the field by analyzing their strengths and weaknesses, performance as well as future prospects Presents appropriate analytical methods and testing procedures in addition to potential device applications Provides strategic insights for those working on diverse application areas such as R&D, regulatory management, and commercial development
Fluorescence microscopy images can be easily integrated into current video and computer image processing systems. People like visual observation; they like to watch a television or computer screen, and fluorescence techniques are thus becoming more and more popular. Since true in vivo experiments are simple to perform, samples can be directly seen and there is always the possibility of manipulating the samples during the experiments; it is an ideal technique for biology and medicine. Images are obtained by a classical (now called wide-field) fluorescence microscope, a confocal scanning microscope, upright or inverted, with epifluorescence or transmission. Computerized image processing may improve definition, and remove glare and scattered light signal. It also makes it possible to compute ratio images (ratio imaging both in excitation and in emission) or lifetime imaging. Image analysis programs may supply a great deal of additional data of various types, starting with calculations of the number of fluorescent objects, their shapes, brightness, etc. Fluorescence microscopy data may be complemented by classical measurement in the cuvette yr by flow cytometry.
Biomedical optical imaging is a rapidly emerging research area with widespread fundamental research and clinical applications. This book gives an overview of biomedical optical imaging with contributions from leading international research groups who have pioneered many of these techniques and applications. A unique research field spanning the microscopic to the macroscopic, biomedical optical imaging allows both structural and functional imaging. Techniques such as confocal and multiphoton microscopy provide cellular level resolution imaging in biological systems. The integration of this technology with exogenous chromophores can selectively enhance contrast for molecular targets as well as supply functional information on processes such as nerve transduction. Novel techniques integrate microscopy with state-of-the-art optics technology, and these include spectral imaging, two photon fluorescence correlation, nonlinear nanoscopy; optical coherence tomography techniques allow functional, dynamic, nanoscale, and cross-sectional visualization. Moving to the macroscopic scale, spectroscopic assessment and imaging methods such as fluorescence and light scattering can provide diagnostics of tissue pathology including neoplastic changes. Techniques using light diffusion and photon migration are a means to explore processes which occur deep inside biological tissues and organs. The integration of these techniques with exogenous probes enables molecular specific sensitivity.
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 choose some of those revolutionary ideas and serve a general audience from broad disciplines to achieve a fundamental understanding of these technologies and to better apply them in their daily research. The book begins with coverage of super-resolution optical microscopy, which discusses targeted modulation such as STED and SIM or localization methods such as PALM. It then discusses novel development of fluorescent probes, such as organic small-molecule probes, fluorescent proteins, and inorganic labels such as quantum dots. Finally, it describes advanced optical microscopy, such as fluorescence lifetime imaging, fiber optic microscopy, scanning ion conductance microscopy, and the joining of optics and acoustics—photoacoustic microscopy. Following each chapter, a detailed list of references is provided. Problems at the end of each chapter are also included.
Supported with 119 illustrations, this milestone work discusses key optical imaging techniques in self-contained chapters; describes the integration of optical imaging techniques with other modalities like MRI, X-ray imaging, and PET imaging; provides a software platform for multimodal integration; presents cutting-edge computational and data processing techniques that ensure rapid, cost-effective, and precise quantification and characterization of the clinical data; covers advances in photodynamic therapy and molecular imaging, and reviews key clinical studies in optical imaging along with regulatory and business issues.
Providing much-needed information on fluorescence spectroscopy and microscopy, this ready reference covers detection techniques, data registration, and the use of spectroscopic tools, as well as new techniques for improving the resolution of optical microscopy below the resolution gap. Starting with the basic principles, the book goes on to treat fluorophores and labeling, single-molecule fluorescence spectroscopy and enzymatics, as well as excited state energy transfer, and super-resolution fluorescence imaging. Examples show how each technique can help in obtaining detailed and refined information from individual molecular systems.
Issues in Nanotechnology / 2012 Edition is a ScholarlyBrief™ that delivers timely, authoritative, comprehensive, and specialized information about Biointerphases in a concise format. The editors have built Issues in Nanotechnology: 2012 Edition on the vast information databases of ScholarlyNews.™ You can expect the information about Biointerphases in this eBook to be deeper than what you can access anywhere else, as well as consistently reliable, authoritative, informed, and relevant. The content of Issues in Nanotechnology / 2012 Edition has been produced by the world’s leading scientists, engineers, analysts, research institutions, and companies. All of the content is from peer-reviewed sources, and all of it is written, assembled, and edited by the editors at ScholarlyEditions™ and available exclusively from us. You now have a source you can cite with authority, confidence, and credibility. More information is available at http://www.ScholarlyEditions.com/.
Molecular imaging is primarily about the chemistry of novelbiological probes, yet the vast majority of practitioners are notchemists or biochemists. This is the first book, written from achemist's point of view, to address the nature of the chemicalinteraction between probe and environment to help elucidatebiochemical detail instead of bulk anatomy. Covers all of the fundamentals of modern imaging methodologies,including their techniques and application within medicine andindustry Focuses primarily on the chemistry of probes and imagingagents, and chemical methodology for labelling andbioconjugation First book to investigate the chemistry of molecularimaging Aimed at students as well as researchers involved in the areaof molecular imaging
Understanding how cells function at the molecular level is the major goal of modern biological research. Fluorescence microscopy provides a direct means of visualizing cellular structures, enzymatic reactions, chromosomal interactions as well as gene regulation at a single-cell level. An obstacle in microscopy is diffraction; which limits the resolution of conventional fluorescence microscopy to ~200 nm laterally and ~500 nm axially. More recently, researchers have developed new modes of microscopy that provide "super-resolution", that is, they circumvent the diffraction limit. Using these new imaging techniques, we can now resolve structures at resolutions that are an order of magnitude improvement over the diffraction limit. This thesis presents several new imaging tools and methods we have developed in our lab to facilitate specific biological studies at high resolution at the single-cell level. Since the invention of super-resolution imaging techniques, the development of labeling methods for fluorescent imaging of non-fluorescent cellular structures has been a rate-limiting step. Here, I describe a novel labeling method developed for super-resolution imaging of cellular structures that utilizes RNA aptamers for labeling proteins with super-resolution compatible dyes. The aptamer tightly binds to a common protein tag - green fluorescent protein (GFP). Since there are so many cell-lines already created that express GFP tagged proteins, this new labeling method can be used to study many proteins of interest without selecting and verifying the binding of a new aptamer to the protein we want to study. I demonstrate the feasibility of this new labeling method by imaging proteins on the plasma membrane as well as proteins inside cells. Another technique we are developing is for use in studies of chromosomal interactions. There are assumptions about how genes are regulated through enhancer-promoter interactions; however, these assumptions are difficult to verify due to difficulties in visualizing the specific processes involved and difficulties in accurately identifying the regulatory sequences and proteins involved. In our new method, we combine locus-specific fluorescent labeling, 2-photon localized crosslinking and sequencing to help us achieve the goal of investigating chromosomal interactions. Lastly, I investigated the use of NADH imaging to studying how a cancer cell's metabolic state may change as the cell squeezes through confined regions in a model system of cancer cell extravasation, which will provide new insights into cancer cell metastasis. In the study, I use microfluidic devices with constrictions of varying widths to mimic the type of constrictions cells experience as they leave and enter vessels. Since Nicotinamide adenine dinucleotide (NADH) is a cofactor involved in many metabolic reactions and is auto-fluorescent when excited with UV light (or 700 nm two-photon excitation), fluorescence lifetime and fluorescence anisotropy imaging techniques were used to monitor the metabolic state of the cells by measuring cellular NADH levels.
Nanotechnology for Biomedical Imaging and Diagnostics: FromNanoparticle Design to Clinical Applications reflects upon theincreasing role of nanomaterials in biological and medical imaging,presenting a thorough description of current research as wellas future directions. With contributions from experts innanotechnology and imaging from academia, industry, andhealthcare, this book provides a comprehensive coverage of thefield, ranging from the architectural design of nanomaterials totheir broad imaging applications in medicine. Grouped into three sections, the book: Elucidates all major aspects of nanotechnology andbioimaging Provides comprehensive coverage of the field, ranging from thearchitectural design of nanomaterials to their broad imagingapplications in medicine Written by well-recognized experts in academia, industry, andhealthcare, will be an excellence source of reference With a multidisciplinary approach and a balance of research anddiagnostic topics, this book will appeal to students, scientiests,and healthcare professionals alike
This third edition of a classic text in biological microscopy includes detailed descriptions and in-depth comparisons of parts of the microscope itself, digital aspects of data acquisition and properties of fluorescent dyes, the techniques of 3D specimen preparation and the fundamental limitations, and practical complexities of quantitative confocal fluorescence imaging. Coverage includes practical multiphoton, photodamage and phototoxicity, 3D FRET, 3D microscopy correlated with micro-MNR, CARS, second and third harmonic signals, ion imaging in 3D, scanning RAMAN, plant specimens, practical 3D microscopy and correlated optical tomography.
One of the major challenges of modern biology and medicine consists in finding means to visualize biomolecules in their natural environment with the greatest level of accuracy, so as to gain insight into their properties and behaviour in a physiological and pathological setting. This has been achieved thanks to the design of novel imaging agents, in particular to fluorescent biosensors. Fluorescence Biosensors comprise a large set of tools which are useful for fundamental purposes as well as for applications in biomedicine, drug discovery and biotechnology. These tools have been designed and engineered thanks to the combined efforts of chemists and biologists over the last decade, and developed hand in hand together with imaging technologies. This volume will convey the many exciting developments the field of fluorescent biosensors and reporters has witnessed over the recent years, from concepts to applications, including chapters on the chemistry of fluorescent probes, on technologies for monitoring protein/protein interactions and technologies for imaging biosensors in cultured cells and in vivo. Other chapters are devoted to specific examples of genetically-encoded reporters, or to protein and peptide biosensors, together with examples illustrating their application to cellular and in vivo imaging, biomedical applications, drug discovery and high throughput screening. Contributions from leading authorities Informs and updates on all the latest developments in the field