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Fluorescence Microscopy of Living Cells in Culture, Part A
Fluorescence Microscopy of Living Cells in Culture, Part B
Recent advances in imaging technology reveal, in real time and great detail, critical changes in living cells and organisms. This manual is a compendium of emerging techniques, organized into two parts: specific methods such as fluorescent labeling, and delivery and detection of labeled molecules in cells; and experimental approaches ranging from the detection of single molecules to the study of dynamic processes in organelles, organs, and whole animals. Although presented primarily as a laboratory manual, the book includes introductory and background material and could be used as a textbook in advanced courses. It also includes a DVD containing movies of living cells in action, created by investigators using the imaging techniques discussed in the book. The editors, David Spector and Robert Goldman, whose previous book was Cells: A Laboratory Manual,are highly respected investigators who have taught microscopy courses at Cold Spring Harbor Laboratory, the Marine Biology Laboratory at Woods Hole, and Northwestern University.
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
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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
In Confocal Microscopy Methods and Protocols, Stephen Paddock and a highly skilled panel of experts lead the researcher using confocal techniques from the bench top, through the imaging process, to the journal page. They concisely describe all the key stages of confocal imaging-from tissue sampling methods, through the staining process, to the manipulation, presentation, and publication of the realized image. Written in a user-friendly, nontechnical style, the methods specifically cover most of the commonly used model organisms: worms, sea urchins, flies, plants, yeast, frogs, and zebrafish. Centered in the many biological applications of the confocal microscope, the book makes possible the successful imaging of both fixed and living specimens using primarily the laser scanning confocal microscope. The powerful hands-on methods collected in Confocal Microscopy Methods and Protocols will help even the novice to produce first-class cover-quality confocal images.
Every cell of the body is dependent on calcium to function. Calcium is found in teeth and bones, and calcium signalling is necessary for the movement of muscles and for the action of the heart and the intestines as well as blood coagulation. Calcium in Living Cells will update classic techniques in detecting microscopic levels of calcium ions (Ca2+) in living cells, as well as address new techniques in the field of calcium detection and calcium signaling. Such detection and measurement of intracellular calcium is important to researchers studying the heart, musculoskeletal, gastrointestinal, and immune systems, whose findings will aid in the advancement of drug and genomic therapies to treat heart, gastrointestinal, autoimmune, and infectious diseases. - Gives researchers much needed information on how to study calcium in live cells, which is becoming increasingly important in heart, musculoskeletal, and immune system research - Provides an overview of the latest methods--fluorescence resonance energy transfer (FRET), for example-- that are new to the field - Allows understanding of how calcium plays a role in intracellular function at the cellular level, which has proved important in Alzheimer's research, heart disease, and areas of musculoskeletal research - Updated chapters reflect advancements in the classic techniques used'preparing calcium buffers, vibrating the Ca2+ Electrode and confocal imaging
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