Quantitative Imaging In Cell Biology

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

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.

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

This book provides an essential overview of existing state-of-the-art quantitative imaging methodologies and protocols (intensity-based ratiometric and FLIM/ PLIM). A variety of applications are covered, including multi-parametric quantitative imaging in intestinal organoid culture, autofluorescence imaging in cancer and stem cell biology, Ca2+ imaging in neural ex vivo tissue models, as well as multi-parametric imaging of pH and viscosity in cancer biology. The current state-of-the-art of 3D tissue models and their compatibility with live cell imaging is also covered. This is an ideal book for specialists working in tissue engineering and designing novel biomaterial.

A diverse collection of state-of-the-art methods for the microscopic imaging of cells and molecules. The authors cover a wide spectrum of complimentary techniques, including such methods as fluorescence microscopy, electron microscopy, atomic force microscopy, and laser scanning cytometry. Additional readily reproducible protocols on confocal scanning laser microscopy, quantitative computer-assisted image analysis, laser-capture microdissection, microarray image scanning, near-field scanning optical microscopy, and reflection contrast microscopy round out this eclectic collection of cutting-edge imaging techniques now available. The authors also discuss preparative methods for particles and cells by transmission electron microscopy.

This detailed volume for the first time explores techniques and protocols involving quantitative imaging flow cytometry (IFC), which has revolutionized our ability to analyze cells, cellular clusters, and populations in a remarkable fashion. Beginning with an introduction to technology, the book continues with sections addressing protocols for studies on the cell nucleus, nucleic acids, and FISH techniques using an IFC instrument, immune response analysis and drug screening, IFC protocols for apoptosis and cell death analysis, as well as morphological analysis and the identification of rare cells. Written for the highly successful Methods in Molecular Biology series, 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 practical, Imaging Flow Cytometry: Methods and Protocols will be a critical source for all laboratories seeking to implement IFC in their research studies.