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Fluorescence imaging, at macro, micro, and submicro scales, has revolutionized biological science in the past 30 years. Immunolabelling has provided precise targeting of molecules in fixed tissue, while fluorescent proteins have enabled localization in living tissues. Fluorescent indicators enable imaging of dynamic changes in cell metabolism. This book covers, for the first time, imaging at all scales from macro to submicro (superresolution). Its authors include Robert Clegg, legendary teacher and researcher (who, sadly, passed away during the editing); Jim Pawley, editor of several editions of the Handbook of Biological Confocal Microscopy; the famous and now dispersed New Zealand team of Mark Cannell, Christian Soeller, and David Baddeley; Robert Hoffman, pioneer of whole-animal imaging in cancer research; Andreas Schoenle and Christian Eggeling on STED nanoscopy, and many more famous participants in this field. All the contributors are at the cutting edge of their field.
This book starts at an introductory level and leads reader to the most advanced topics in fluorescence imaging and super-resolution techniques that have enabled new developments such as nanobioimaging, multiphoton microscopy, nanometrology and nanosensors. The interdisciplinary subject of fluorescence microscopy and imaging requires complete knowledge of imaging optics and molecular physics. So, this book approaches the subject by introducing optical imaging concepts before going in more depth about advanced imaging systems and their applications. Additionally, molecular orbital theory is the important basis to present molecular physics and gain a complete understanding of light-matter interaction at the geometrical focus. The two disciplines have some overlap since light controls the molecular states of molecules and conversely, molecular states control the emitted light. These two mechanisms together determine essential imaging factors such as, molecular cross-section, Stoke shift, emission and absorption spectra, quantum yield, signal-to-noise ratio, Forster resonance energy transfer (FRET), fluorescence recovery after photobleaching (FRAP) and fluorescence lifetime. These factors form the basis of many fluorescence based devices. The book is organized into two parts. The first part deals with basics of imaging optics and its applications. The advanced part takes care of several imaging techniques and related instrumentation that are developed in the last decade pointing towards far-field diffraction unlimited imaging.
Covers three main areas.The phenomenon of fluorescence, the main applications, and the most common problems.
The third edition of this established classic text reference builds upon the strengths of its very popular predecessors. Organized as a broadly useful textbook Principles of Fluorescence Spectroscopy, 3rd edition maintains its emphasis on basics, while updating the examples to include recent results from the scientific literature. The third edition includes new chapters on single molecule detection, fluorescence correlation spectroscopy, novel probes and radiative decay engineering. Includes a link to Springer Extras to download files reproducing all book artwork, for easy use in lecture slides. This is an essential volume for students, researchers, and industry professionals in biophysics, biochemistry, biotechnology, bioengineering, biology and medicine.
Fluorescence imaging, at macro, micro, and submicro scales, has revolutionized biological science in the past 30 years. Immunolabelling has provided precise targeting of molecules in fixed tissue, while fluorescent proteins have enabled localization in living tissues. Fluorescent indicators enable imaging of dynamic changes in cell metabolism. This book covers, for the first time, imaging at all scales from macro to submicro (superresolution). Its authors include Robert Clegg, legendary teacher and researcher (who, sadly, passed away during the editing); Jim Pawley, editor of several editions of the Handbook of Biological Confocal Microscopy; the famous and now dispersed New Zealand team of Mark Cannell, Christian Soeller, and David Baddeley; Robert Hoffman, pioneer of whole-animal imaging in cancer research; Andreas Schoenle and Christian Eggeling on STED nanoscopy, and many more famous participants in this field. All the contributors are at the cutting edge of their field.
In the last decade, fluorescence microscopy has evolved from a classical “retrospective” microscopy approach into an advanced imaging technique that allows the observation of cellular activities in living cells with increased resolution and dimensions. A bright new future has arrived as the nano era has placed a whole new array of tools in the hands of biophysicists who are keen to go deeper into the intricacies of how biological systems work. Following an introduction to the complex world of optical microscopy, this book covers topics such as the concept of white confocal, nonlinear optical microscopy, fluctuation spectroscopies, site-specific labeling of proteins in living cells, imaging molecular physiology using nanosensors, measuring molecular dynamics, muscle braking and stem cell differentiation.
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
A self-contained treatment of the latest fluorescence applications in biotechnology and the life sciences This book focuses specifically on the present applications of fluorescence in molecular and cellular dynamics, biological/medical imaging, proteomics, genomics, and flow cytometry. It raises awareness of the latest scientific approaches and technologies that may help resolve problems relevant for the industry and the community in areas such as public health, food safety, and environmental monitoring. Following an introductory chapter on the basics of fluorescence, the book covers: labeling of cells with fluorescent dyes; genetically encoded fluorescent proteins; nanoparticle fluorescence probes; quantitative analysis of fluorescent images; spectral imaging and unmixing; correlation of light with electron microscopy; fluorescence resonance energy transfer and applications; monitoring molecular dynamics in live cells using fluorescence photo-bleaching; time-resolved fluorescence in microscopy; fluorescence correlation spectroscopy; flow cytometry; fluorescence in diagnostic imaging; fluorescence in clinical diagnoses; immunochemical detection of analytes by using fluorescence; membrane organization; and probing the kinetics of ion pumps via voltage-sensitive fluorescent dyes. With its multidisciplinary approach and excellent balance of research and diagnostic topics, this book is an essential resource for postgraduate students and a broad range of scientists and researchers in biology, physics, chemistry, biotechnology, bioengineering, and medicine.
In color throughout, this text helps readers acquire a sound understanding of basic fluorescence theory and practice. It takes them through the history of important discoveries to the most current advances. The author introduces the fundamentals of the fluorescence phenomenon and gives detailed examples of fluorescence applications in the molecular life sciences, including biochemistry, biophysics, clinical chemistry and diagnostics, pharmaceutical science, and cell and molecular biology. The text includes references in each chapter, more than 250 figures, and the chemical structures of the most widely used fluorescent molecules.