This new volume, number 123, of Methods in Cell Biology looks at methods for quantitative imaging in cell biology. It covers both theoretical and practical aspects of using optical fluorescence microscopy and image analysis techniques for quantitative applications. The introductory chapters cover fundamental concepts and techniques important for obtaining accurate and precise quantitative data from imaging systems. These chapters address how choice of microscope, fluorophores, and digital detector impact the quality of quantitative data, and include step-by-step protocols for capturing and analyzing quantitative images. Common quantitative applications, including co-localization, ratiometric imaging, and counting molecules, are covered in detail. Practical chapters cover topics critical to getting the most out of your imaging system, from microscope maintenance to creating standardized samples for measuring resolution. Later chapters cover recent advances in quantitative imaging techniques, including super-resolution and light sheet microscopy. With cutting-edge material, this comprehensive collection is intended to guide researchers for years to come. Covers sections on model systems and functional studies, imaging-based approaches and emerging studies Chapters are written by experts in the field Cutting-edge material

Fluorescence Microscopy of Living Cells in Culture, Part B

Quantitative fluorescence microscopy is concerned with making measurements from fluorescent specimens in a fluorescence microscope, by measuring fluorescence emission from a defined area or areas of a specimen. This technique is most commonly used to determine the amount of some specific substance, such as DNA, in some particular area of a cell. But it has many other uses; for example, it can be used to identify certain substances in the cell by examining their fluorescence characteristics. This book is a complete guide to this technique for all biologists. It describes the principles and applications of quantitative fluorescence microscopy and also gives much practical information about the instrumentation required. There is also a discussion of the exciting developments in confocal fluorescence microscopy which allows the three dimensional distribution of particular substances to be determined. Everyone presently using this technique, or wishing to start using it will need to read this book.

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.

Quantitative bioimaging is a broad interdisciplinary field that exploits tools from biology, chemistry, optics, and statistical data analysis for the design and implementation of investigations of biological processes. Instead of adopting the traditional approach of focusing on just one of the component disciplines, this textbook provides a unique introduction to quantitative bioimaging that presents all of the disciplines in an integrated manner. The wide range of topics covered include basic concepts in molecular and cellular biology, relevant aspects of antibody technology, instrumentation and experimental design in fluorescence microscopy, introductory geometrical optics and diffraction theory, and parameter estimation and information theory for the analysis of stochastic data. Key Features: Comprises four parts, the first of which provides an overview of the topics that are developed from fundamental principles to more advanced levels in the other parts. Presents in the second part an in-depth introduction to the relevant background in molecular and cellular biology and in physical chemistry, which should be particularly useful for students without a formal background in these subjects. Provides in the third part a detailed treatment of microscopy techniques and optics, again starting from basic principles. Introduces in the fourth part modern statistical approaches to the determination of parameters of interest from microscopy data, in particular data generated by single molecule microscopy experiments. Uses two topics related to protein trafficking (transferrin trafficking and FcRn-mediated antibody trafficking) throughout the text to motivate and illustrate microscopy techniques. An online appendix providing the background and derivations for various mathematical results presented or used in the text is available at http://www.routledge.com/9781138598980.

This book offers a comprehensive selection of essays by leading experts, which covers all aspects of modern imaging, from its application and up-scaling to its development. The chapter content ranges from the basics to the most complex overview of method and protocols. There is ample practical and detailed "how-to" content on important, but rarely addressed topics. This first edition features all-colour-plate chapters, licensed software and a unique, continuously updated website forum.