Imaging Living Cells

This detailed book highlights recent advances in molecular imaging techniques and protocols, designed to be immediately applicable in global bio-laboratories. The chapters are categorized into seven major groups according to the reporter materials, such as imaging with passive optical readouts, activatable bioluminescent probes, functional substrates and luciferases, organic fluorescent probes, BRET probes, FRET probes, as well as with advanced instrumentation. 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, Live Cell Imaging: Methods and Protocols aims to direct and inspire researchers into creating smarter, next-generation imaging techniques that are truly quantitative, highly sensitive, and readily comprehended, in the effort to engender deeper understanding of biological systems and break new ground in the research fields of life science.

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.

This volume of Methods in Enzymology is the second of three parts looking at current methodology for the imaging and spectroscopic analysis of live cells. The chapters provide hints and tricks not available in primary research publications. It is an invaluable resource for academics, researchers and students alike. Expert authors who are leaders in the field Extensively referenced and useful figures and tables Provides hints and tricks to facilitate reproduction of methods

This volume of Methods in Enzymology is the third of 3 parts looking at current methodology for the imaging and spectroscopic analysis of live cells. The chapters provide hints and tricks not available in primary research publications. It is an invaluable resource for academics, researchers and students alike. Publisher's note.

Intended for bench-top use, this lab manual is suitable for both scientists and graduate students, since it combines an update on the most advanced imaging procedures with detailed protocols. Examples, carefully selected from the wide repertoire of cell pyhsiology, cover such different functional aspects as distribution of multiple ions, electrical activity, exo-endocytosis, gene expression, and the cell cycle.

Fluorescence Microscopy of Living Cells in Culture, Part B

This volume of Methods in Enzymology is the first of three parts looking at current methodology for the imaging and spectroscopic analysis of live cells. The chapters provide hints and tricks not available in primary research publications. It is an invaluable resource for academics, researchers and students alike. Expert authors who are leaders in the field Extensively referenced and useful figures and tables Provides hints and tricks to facilitate reproduction of methods

Advances in technology have revolutionized the development of light microscopy techniques in biomedical research, thus improving visualization of the microstructure of cells and tissues under physiological conditions. Fluorescence microscopy methods are non-contact and non-invasive and provide high spatial and temporal resolution that other laboratory techniques cannot. This well-illustrated book targets graduate students and scientists who are new to the state-of-the-art fluorescence microscopy techniques used in biological and clinical imaging. It explains basic concepts and imaging procedures for wide-field, confocal, multiphoton excitation, fluorescence resonance energy transfer (FRET), lifetime imaging (FLIM), spectral imaging, fluorescence recovery after photobleaching (FRAP), optical tweezers, total internal reflection, high spatial resolution atomic force microscopy (AFM), and bioluminescence imaging for gene expression. The usage of these techniques in various biological applications, including calcium, pH, membrane potential, mitochondrial signaling, protein-protein interactions under various physiological conditions, and deep tissue imaging, is clearly presented. The authors describe the approaches to selecting epifluorescence microscopy, the detectors, and the image acquisition and processing software for different biological applications. Step-by-step directions on preparing different digital formats for light microscopy images on websites are also provided.