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.

"Alberto Diaspro has been choreographing light's dance for over 20 years, and in Nanoscopy and Multidimensional Optical Fluorescence Microscopy, he has assembled a diverse group of experts to explain the methods they use to coax light to reveal biology's secrets."- From the Foreword by Daniel Evanko, editor, Nature Methods Nanoscopy and Multidimens

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.

This book provides a comprehensive account of the theory of image formation in a confocal fluorescence microscope as well as a practical guideline to the operation of the instrument, its limitations, and the interpretation of confocal microscopy data. The appendices provide a quick reference to optical theory, microscopy-related formulas and definitions, and Fourier theory.

Covers three main areas.The phenomenon of fluorescence, the main applications, and the most common problems.

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.

Presents a fully updated, self-contained textbook covering the core theory and practice of both classical and modern optical microscopy techniques.

The purpose of this book is to provide the most comprehensive, easy-to-use, and informative guide on light microscopy. Light and Video Microscopy will prepare the reader for the accurate interpretation of an image and understanding of the living cell. With the presentation of geometrical optics, it will assist the reader in understanding image formation and light movement within the microscope. It also provides an explanation of the basic modes of light microscopy and the components of modern electronic imaging systems and guides the reader in determining the physicochemical information of living and developing cells, which influence interpretation. - Brings together mathematics, physics, and biology to provide a broad and deep understanding of the light microscope - Clearly develops all ideas from historical and logical foundations - Laboratory exercises included to assist the reader with practical applications - Microscope discussions include: bright field microscope, dark field microscope, oblique illumination, phase-contrast microscope, photomicrography, fluorescence microscope, polarization microscope, interference microscope, differential interference microscope, and modulation contrast microscope

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.

Choice Recommended Title, March 2020 Optical microscopy is used in a vast range of applications ranging from materials engineering to in vivo observations and clinical diagnosis, and thanks to the latest advances in technology, there has been a rapid growth in the number of methods available. This book is aimed at providing users with a practical guide to help them select, and then use, the most suitable method for their application. It explores the principles behind the different forms of optical microscopy, without the use of complex maths, to provide an understanding to help the reader utilise a specific method and then interpret the results. Detailed physics is provided in boxed sections, which can be bypassed by the non-specialist. It is an invaluable tool for use within research groups and laboratories in the life and physical sciences, acting as a first source for practical information to guide less experienced users (or those new to a particular methodology) on the range of techniques available. Features: The first book to cover all current optical microscopy methods for practical applications Written to be understood by a non-optical expert with inserts to provide the physical science background Brings together conventional widefield and confocal microscopy, with advanced non-linear and super resolution methods, in one book To learn more about the author please visit here.