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

Methods in Neurosciences, Volume 3: Quantitative and Qualitative Microscopy is a collection of papers that deals with microscopic techniques in statistical measures. This volume describes microscopy using sophisticated stains and dyes to advance observation of tests and experiments. Section I describes autoradiography including micro chemical methods, high-resolution autoradiography, and single- or double-label quantitative autoradiography for use in imaging of brain activity patterns or determining cerebral physiology. Section II discusses the quantification of structures through statistical and computational methods including dynamic video imaging technology. Section III explains the use of tracers, toxins, or dyes in tracing neuronal connections. One paper addresses the use of small injections of axonally transported fluorescent tracers. Section IV explains staining technology such as using the silver impregnation method for frozen sections of human nervous tissue that are gathered from tissues preserved in formalin. Section V addresses freezing techniques and those using freeze-fracture methods in neurobiology. The text also discusses cryoprotection and other freezing methods to control ice crystals found in fixed or unfixed brain tissues. Section VI presents the combined and high-resolution methods in polarization microscopy and microscopic investigations. Cellular biologists, micro-chemists, and scientific researchers in the field of micro- and cellular biology will appreciate this book.

This book deals with the latest achievements in the field of optical coherent microscopy. While many other books exist on microscopy and imaging, this book provides a unique resource dedicated solely to this subject. Similarly, many books describe applications of holography, interferometry and speckle to metrology but do not focus on their use for microscopy. The coherent light microscopy reference provided here does not focus on the experimental mechanics of such techniques but instead is meant to provide a users manual to illustrate the strengths and capabilities of developing techniques. The areas of application of this technique are in biomedicine, medicine, life sciences, nanotechnology and materials sciences.

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.

Fluorescence Microscopy of Living Cells in Culture, Part B

Cutting-edge quantitative phase imaging techniques and their applications Filled with unique, full-color images taken by advanced quantitative phase imaging (QPI), Quantitative Phase Imaging of Cells and Tissues thoroughly explores this innovative technology and its biomedical applications. An introductory background on optical imaging and traditional optical microscopy is included to illustrate concept development. The book explains how various visualization modalities can be obtained by numerical calculations. This authoritative resource reveals how to take full advantage of the unprecedented capabilities of QPI, such as rendering scattering properties of minute subcellular structures and nanoscale fluctuations in live cells. Coverage includes: Groundwork Spatiotemporal field correlations Image characteristics Light microscopy Holography Point scanning QPI methods Principles of full-field QPI Off-axis full-field methods Phase-shifting techniques Common-path methods White light techniques Fourier transform light scattering (FTLS) Current trends in QPI

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