This book offers an overview of imaging techniques used to investigate cells and tissue in their native environment. It covers the range of imaging approaches used, as well as the application of those techniques to the study of biological processes in cells and whole tissues within living organisms.
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.
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 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
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.
Bio-Nanoimaging: Protein Misfolding & Aggregation provides a unique introduction to both novel and established nanoimaging techniques for visualization and characterization of misfolded and aggregated protein species. The book is divided into three sections covering: - Nanotechnology and nanoimaging technology, including cryoelectron microscopy of beta(2)-microglobulin, studying amyloidogensis by FRET; and scanning tunneling microscopy of protein deposits - Polymorphisms of protein misfolded and aggregated species, including fibrillar polymorphism, amyloid-like protofibrils, and insulin oligomers - Polymorphisms of misfolding and aggregation processes, including multiple pathways of lysozyme aggregation, misfolded intermediate of a PDZ domain, and micelle formation by human islet amyloid polypeptide Protein misfolding and aggregation is a fast-growing frontier in molecular medicine and protein chemistry. Related disorders include cataracts, arthritis, cystic fibrosis, late-onset diabetes mellitus, and numerous neurodegenerative diseases like Alzheimer's and Parkinson's. Nanoimaging technology has proved crucial in understanding protein-misfolding pathologies and in potential drug design aimed at the inhibition or reversal of protein aggregation. Using these technologies, researchers can monitor the aggregation process, visualize protein aggregates and analyze their properties. - Provides practical examples of nanoimaging research from leading molecular biology, cell biology, protein chemistry, biotechnology, genetics, and pharmaceutical labs - Includes over 200 color images to illustrate the power of various nanoimaging technologies - Focuses on nanoimaging techniques applied to protein misfolding and aggregation in molecular medicine
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
These are exciting times for the field of optical imaging of brain function. Rapid developments in theory and technology continue to considerably advance understanding of brain function. Reflecting changes in the field during the past five years, the second edition of In Vivo Optical Imaging of Brain Function describes state-of-the-art techniques and their applications for the growing field of functional imaging in the live brain using optical imaging techniques. New in the Second Edition: Voltage-sensitive dyes imaging in awake behaving animals Imaging based on genetically encoded probes Imaging of mitochondrial auto-fluorescence as a tool for cortical mapping Using pH-sensitive dyes for functional mapping Modulated imaging Calcium imaging of neuronal activity using 2-photon microscopy Fourier approach to optical imaging Fully updated chapters from the first edition Leading Authorities Explore the Latest Techniques Updated to reflect continuous development in this emerging research area, this new edition, as with the original, reaches across disciplines to review a variety of non-invasive optical techniques used to study activity in the living brain. Leading authorities from such diverse areas as biophysics, neuroscience, and cognitive science present a host of perspectives that range from a single neuron to large assemblies of millions of neurons, captured at various temporal and spatial resolutions. Introducing techniques that were not available just a few years ago, the authors describe the theory, setup, analytical methods, and examples that highlight the advantages of each particular method.
Cutting-edge quantitative phase imaging techniques and their applications Filled with unique, full-color images taken by advanced quantitative phase imaging (QPI), Quantitative Phase Imaging of Cells and Tissues thoroughly explores this innovative technology and its biomedical applications. An introductory background on optical imaging and traditional optical microscopy is included to illustrate concept development. The book explains how various visualization modalities can be obtained by numerical calculations. This authoritative resource reveals how to take full advantage of the unprecedented capabilities of QPI, such as rendering scattering properties of minute subcellular structures and nanoscale fluctuations in live cells. Coverage includes: Groundwork Spatiotemporal field correlations Image characteristics Light microscopy Holography Point scanning QPI methods Principles of full-field QPI Off-axis full-field methods Phase-shifting techniques Common-path methods White light techniques Fourier transform light scattering (FTLS) Current trends in QPI