Fundamentals of Fluorescence Imaging


Book Description

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.




Fundamentals of Fluorescence Microscopy


Book Description

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.







Fundamentals of Fluorescence Imaging


Book Description

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.




Principles of Fluorescence Spectroscopy


Book Description

Fluorescence methods are being used increasingly in biochemical, medical, and chemical research. This is because of the inherent sensitivity of this technique. and the favorable time scale of the phenomenon of fluorescence. 8 Fluorescence emission occurs about 10- sec (10 nsec) after light absorp tion. During this period of time a wide range of molecular processes can occur, and these can effect the spectral characteristics of the fluorescent compound. This combination of sensitivity and a favorable time scale allows fluorescence methods to be generally useful for studies of proteins and membranes and their interactions with other macromolecules. This book describes the fundamental aspects of fluorescence. and the biochemical applications of this methodology. Each chapter starts with the -theoreticalbasis of each phenomenon of fluorescence, followed by examples which illustrate the use of the phenomenon in the study of biochemical problems. The book contains numerous figures. It is felt that such graphical presentations contribute to pleasurable reading and increased understand ing. Separate chapters are devoted to fluorescence polarization, lifetimes, quenching, energy transfer, solvent effects, and excited state reactions. To enhance the usefulness of this work as a textbook, problems are included which illustrate the concepts described in each chapter. Furthermore, a separate chapter is devoted to the instrumentation used in fluorescence spectroscopy. This chapter will be especially valuable for those perform ing or contemplating fluorescence measurements. Such measurements are easily compromised by failure to consider a number of simple principles.




Optical Fluorescence Microscopy


Book Description

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.




Fluorescence Microscopy in Life Sciences


Book Description

Fluorescence Microscopy is a precise and widely employed technique in many research and clinical areas nowadays. Fluorescence Microscopy In Life Sciences introduces readers to both the fundamentals and the applications of fluorescence microscopy in the biomedical field as well as biological research. Readers will learn about physical and chemical mechanisms giving rise to the phenomenon of luminescence and fluorescence in a comprehensive way. Also, the different processes that modulate fluorescence efficiency and fluorescence features are explored and explained.




Fundamentals of Light Microscopy and Electronic Imaging


Book Description

Fundamentals of Light Microscopy and Electronic Imaging, Second Edition provides a coherent introduction to the principles and applications of the integrated optical microscope system, covering both theoretical and practical considerations. It expands and updates discussions of multi-spectral imaging, intensified digital cameras, signal colocalization, and uses of objectives, and offers guidance in the selection of microscopes and electronic cameras, as well as appropriate auxiliary optical systems and fluorescent tags. The book is divided into three sections covering optical principles in diffraction and image formation, basic modes of light microscopy, and components of modern electronic imaging systems and image processing operations. Each chapter introduces relevant theory, followed by descriptions of instrument alignment and image interpretation. This revision includes new chapters on live cell imaging, measurement of protein dynamics, deconvolution microscopy, and interference microscopy. PowerPoint slides of the figures as well as other supplementary materials for instructors are available at a companion website: www.wiley.com/go/murphy/lightmicroscopy




Introduction to Fluorescence


Book Description

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.




Fluorescence Microscopy


Book Description

This book covers the fundamental principles of fluorescence and their application to fluorescence microscopy, and presents applications to immunofluorescence, in situ hybridization, and photomicrography. It provides troubleshooting guidance to guide the user through commonly encountered problems.