Cell Imaging

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.

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 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

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.

This book focuses on the latest fluorescent materials for cell imaging. Cell imaging is a widely used basic technique that helps scientists gain a better understanding of biological functions through studies of cellular structure and dynamics. In the past decades, the development of a variety of new fluorescent materials has significantly extended the applications of cellular imaging techniques. This book presents recently developed fluorescent materials, including semiconductor quantum dots, carbon dots, silicon nanoparticles, metal nanoclusters, upconversion nanoparticles, conjugated polymers/polymer dots, aggregation-induced emission (AIE) probes, and coordination compounds, used for various cellular imaging purposes. It will appeal to cell biologists and other researchers in academia, industry and clinical settings who are interested in the technical development and advanced applications of fluorescence imaging in cells, tissues and organisms to explore the mechanisms of biological functions and diseases.

In the biomedical sciences, the confocal laser scanning microscope (CLSM) has become the instrument of choice for producing high-resolution images and 3D reconstruction, breaking the barriers of conventional optical microscopy. Wouterlood (anatomy, VU U. Medical Center, Amsterdam, the Netherlands) introduces the confocal principle which eliminates out-of-focus haze, its components, and relevant equations. International scientists explain the principles and related methods of stimulated emission depletion (SRED), single molecule localization, and coherent anti-Stokes Raman (CARS) microscopy; labeling approaches; preparation of samples for imaging; and applications of, and developments in, this new wave of imaging, e.g., visualization of neuronal networks, DNA, and myelin. The text includes color and b&w images, and referral to an online CLSM simulator. Academic Press is an imprint of Elsevier. Annotation ©2013 Book News, Inc., Portland, OR (booknews.com).

This book highlights important techniques for cellular imaging and covers the basics and applications of electron tomography and related techniques. In addition, it considers practical aspects and broadens the technological focus by incorporating techniques that are only now becoming accessible (e.g. block face imaging). The first part of the book describes the electron microscopy 3D technique available to scientists around the world, allowing them to characterize organelles, cells and tissues. The major emphasis is on new technologies like scanning transmission electron microscopy (STEM) tomography, though the book also reviews some of the more proven technologies like electron tomography. In turn, the second part is dedicated to the reconstruction of data sets, signal improvement and interpretation