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 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

There continues to be intense interest in the microtubule cytoskeleton; the assembly, structure and regulation of microtubules; and the numerous motors and accessory proteins that control cell cycle, dynamics, organization and transport. The field continues to grow and explore new aspects of these issues driven immensely by developments in optical imaging and tracking techniques. This Second Edition brings together current research and protocols in the field of microtubules in vitro and will serve as a valuable tool for cell biologists, biophysicists and pharmacologists who study the microtubule cytoskeleton, as well as for researchers in the biomedical and biotechnology communities with interest in developing drugs that target microtubules, MAPS and motors. - Chapters reflect experimental procedures and new developments in the field of microtubule in vitro research - Combines classical approaches and modern technologies - Presents easy-to-use protocols and thorough background information, compiled by leaders in the field

Computational methods are playing an ever increasing role in cell biology. This volume of Methods in Cell Biology focuses on Computational Methods in Cell Biology and consists of two parts: (1) data extraction and analysis to distill models and mechanisms, and (2) developing and simulating models to make predictions and testable hypotheses. - Focuses on computational methods in cell biology - Split into 2 parts--data extraction and analysis to distill models and mechanisms, and developing and simulating models to make predictions and testable hypotheses - Emphasizes the intimate and necessary connection with interpreting experimental data and proposing the next hypothesis and experiment

Cell biology spans among the widest diversity of methods in the biological sciences. From physical chemistry to microscopy, cells have given up with secrets only when the questions are asked in the right way! This new volume of Methods in Cell Biology covers laboratory methods in cell biology, and includes methods that are among the most important and elucidating in the discipline, such as transfection, cell enrichment and magnetic batch separation. Covers the most important laboratory methods in cell biology Chapters written by experts in their fields.

The first of its kind, this laboratory handbook emphasizes diverse methods and technologies needed to investigate C. elegans, both as an integrated organism and as a model system for research inquiries in cell, developmental, and molecular biology, as well as in genetics and pharmacology. Four primary sections--Genetic and Culture Methods, Neurobiology, Cell and Molecular Biology, and Genomics and Informatics--reflect the cross-disciplinary nature of C. elegans research. Because C. elegans is a simple and malleable organism with a small genome and few cell types, it provides an elegant demonstr.

Methods in Cell Biology, Volume 158, the latest release in this series, highlights new advances in the field, with this release covering How to orient cells in micro-cavities for high resolution imaging of cytokinesis and lumen formation, A body-on-a-chip (BOC) system for studying gut-liver interaction, Manipulating cultured mammalian cells for mitosis research, Live-cell FLIM-FRET using a commercially available system, A comparative analysis of methods to measure kinetochore-microtubule attachment stability, A workflow for visualizing human cancer biopsies using large-format electron microscopy, Isolation of stage-specific germ cells using facs in drosophila germarium, Computational analysis of filament polymerization dynamics in cytoskeletal networks, and more. Provides the authority and expertise of leading contributors from an international board of authors Presents the latest release in the Methods in Cell Biology series Updated release includes the latest information in this area of study

Recent advances in the imaging technique electron microscopy (EM) have improved the method, making it more reliable and rewarding, particularly in its description of three-dimensional detail. Cellular Electron Microscopy will help biologists from many disciplines understand modern EM and the value it might bring to their own work. The book's five sections deal with all major issues in EM of cells: specimen preparation, imaging in 3-D, imaging and understanding frozen-hydrated samples, labeling macromolecules, and analyzing EM data. Each chapter was written by scientists who are among the best in their field, and some chapters provide multiple points of view on the issues they discuss. Each section of the book is preceded by an introduction, which should help newcomers understand the subject. The book shows why many biologists believe that modern EM will forge the link between light microscopy of live cells and atomic resolution studies of isolated macromolecules, helping us toward the goal of an atomic resolution understanding of living systems. - Updates the numerous technological innovations that have improved the capabilities of electron microscopy - Provides timely coverage of the subject given the significant rise in the number of biologists using light microscopy to answer their questions and the natural limitations of this kind of imaging - Chapters include a balance of "how to", "so what" and "where next", providing the reader with both practical information, which is necessary to use these methods, and a sense of where the field is going

A Practical Guide to the Study of Calcium in Living Cells describes popular techniques along with helpful do's and don't's and computer programs. The volume enables investigators to evaluate confocal images, use the latest dyes, and design Calcium buffers appropriate to their research needs. This book is designed for laboratory use by graduate students, technicians, and researchers in many disciplines, ranging from molecular to cellular levels of investigation. - Describes techniques for detection of [Ca2+]I: Ca2+ - sensitive microelectrodes - Fluorescent dyes - Luminescent proteins - Includes techniques for perturbing intracellular Ca2+ - Covers detailed methodology plus problems and pitfalls of each technique - Contains a practical guide to preparing Ca2+ buffers with an easy-to-use computer program - Color plates illustrate techniques such as - Confocal ratio-imaging - Use of aequorin

Methods in Cell Biology Volume 155 provides an update on the step-by-step "how-to" methods to study mitochondrial structure, function and biogenesis contained in the first two editions. As in the previous editions, biochemical, cell biological, and genetic approaches are presented along with sample results, interpretations, and pitfalls for each method. New chapters in this update include Isolation of Mitochondria and Analysis of Mitochondrial Compartments, Isolation of Mitochondria from Animal Cells and Yeast, Isolation and Characterization of Mitochondria-Associated ER Membranes, Import of Proteins into Mitochondria, Proximity Labeling Methods to Assess Protein-Protein Interactions in Yeast Mitochondria, and more. Provides a step-by-step "cookbook" presentation as written by leaders in the field Covers longstanding methods that have shaped the field Includes the newest technologies and methods