This volume provides an overview of advanced fluorescence microscopy, covering a broad range of methods. Each chapter focuses on a different method and provides a practical guide for application in biological systems. Written in the highly successful Methods in Molecular Biology series format, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and tips on troubleshooting and avoiding known pitfalls. Authoritative and cutting-edge, Advanced Fluorescence Microscopy: Methods and Protocols seeks to provide scientists with methods for biological systems that are of interest.

Zu dem Thema gibt es viele Publikationen, die von Experten für Experten geschrieben wurden. Dieses Buch wendet sich insbesondere an Studenten höherer Semester und Forscher, denen das Hintergrundwissen der Physik fehlt, um neuartige Verfahren der Fluoreszenzmikroskopie zu verstehen. Die zweite Auflage wartet mit neuen Kapiteln und einer erweiterten Einführung auf. Der Schwerpunkt liegt auf der hochauflösenden und Einzelmolekül-Mikroskopie. Jedes Kapitel wurde von einem anerkannten Experten des Fachgebiets geschrieben und sorgfältig überarbeitet, um so die Entwicklungen der letzten Jahre wiederzugeben.

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.

This text draws together the fields of optical microscopy and optical data storage, in a unique compilation of valuable and novel scientific work that is scarcely to be found elsewhere. The contributing authors are unquestioned leaders of their respective fields.

This book is a wide-ranging guide to advanced imaging techniques and related methods with important applications in translational research or convergence science as progress is made toward a new era in integrative healthcare. Conventional and advanced microscopic imaging techniques, including both non-fluorescent (i.e., label-free) and fluorescent methods, have to date provided researchers with specific and quantitative information about molecules, cells, and tissues. Now, however, the different imaging techniques can be correlated with each other and multimodal methods developed to simultaneously obtain diverse and complementary information. In addition, the latest advanced imaging techniques can be integrated with non-imaging techniques such as mass spectroscopic methods, genome editing, organic/inorganic probe synthesis, nanomedicine, and drug discovery. The book will be of high value for researchers in the biological and biomedical sciences or convergence science who need to use these multidisciplinary and integrated techniques or are involved in developing new analytical methods focused on convergence science.

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.

With contributions by numerous experts

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.

Over the last decade, fluorescence has become the dominant tool in biotechnology and medical imaging. These exciting advances have been underpinned by the advances in time-resolved techniques and instrumentation, probe design, chemical / biochemical sensing, coupled with our furthered knowledge in biology. Complementary volumes 9 & 10, Advanced Concepts of Fluorescence Sensing: Small Molecule Sensing and Advanced Concepts of Fluorescence Sensing: Macromolecular Sensing, aim to summarize the current state of the art in fluorescent sensing. For this reason, Drs. Geddes and Lakowicz have invited chapters, encompassing a broad range of fluorescence sensing techniques. Some chapters deal with small molecule sensors, such as for anions, cations, and CO2, while others summarize recent advances in protein-based and macromolecular sensors. The Editors have, however, not included DNA or RNA based sensing in this volume, as this were reviewed in Volume 7 and is to be the subject of a more detailed volume in the near future.