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This book will serve as an introduction to microscopy and biomedical imaging methods, with a focus on the study of the distributions and dynamics of molecules on the cell surface. It will provide readers with an in-depth understanding of how modern microscopy methodology can be used to understand the organisation of cell membrane systems and how experiments can be designed around these methodologies. There are numerous methods employed to understand cell membrane organisation, but foremost among them are microscopy methods which can map the distributions of molecules on the cell surface and even map the biophysical properties of membranes themselves. Fluorescence microscopy has been especially widely used due to its specificity and relatively noninvasive nature, allowing live-cell imaging. However, the recent advance of super-resolution fluorescence microscopy has broken the previous resolution limit for this type of microscopy, which has been an important advancement in the field. Atomic force microscopy and electron microscopy have also been deployed to learn about membrane organisation and properties. Each chapter in this volume will be themed around measuring a particular property of cell membranes. In each case, the authors examine the range of methodology applicable to the task, comparing the advantages and disadvantages of each one, and will also provide an overview of important discoveries that have been made using the methodology being discussed. The chapters will cover: • Measuring membrane protein distributions using single-molecule localisation microscopy (SMLM) • Measuring membrane protein dynamics and diffusion using fluorescence correla-tion spectroscopy (FCS) • Mapping membrane lipid backing using environmentally sensitive fluorescence probes • Mapping membrane thickness and rigidity using atomic force microscopy • Mapping membrane proteins and the cytoskeleton using electron microscopy This book will be a valuable resource to graduate and upper-level undergraduate students and industry researchers in the fields of cell biology, microbiology, microscopy, and medical imaging.
· Measuring membrane protein distributions using single-molecule localisation microscopy (SMLM) · Measuring membrane protein dynamics and diffusion using fluorescence correlation spectroscopy (FCS) · Mapping membrane lipid backing using environmentally sensitive fluorescence probes · Mapping membrane thickness and rigidity using atomic force microscopy · Mapping membrane proteins and the cytoskeleton using electron microscopy
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.
2.6.2 Electrodes for Electrochemistry
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
Cell Membrane Nanodomains: From Biochemistry to Nanoscopy describes recent advances in our understanding of membrane organization, with a particular focus on the cutting-edge imaging techniques that are making these new discoveries possible. With contributions from pioneers in the field, the book explores areas where the application of these novel techniques reveals new concepts in biology. It assembles a collection of works where the integration of membrane biology and microscopy emphasizes the interdisciplinary nature of this exciting field. Beginning with a broad description of membrane organization, including seminal work on lipid partitioning in model systems and the roles of proteins in membrane organization, the book examines how lipids and membrane compartmentalization can regulate protein function and signal transduction. It then focuses on recent advances in imaging techniques and tools that foster further advances in our understanding of signaling nanoplatforms. The coverage includes several diffraction-limited imaging techniques that allow for measurements of protein distribution/clustering and membrane curvature in living cells, new fluorescent proteins, novel Laurdan analyses, and the toolbox of labeling possibilities with organic dyes. Since superresolution optical techniques have been crucial to advancing our understanding of cellular structure and protein behavior, the book concludes with a discussion of technologies that are enabling the visualization of lipids, proteins, and other molecular components at unprecedented spatiotemporal resolution. It also explains the ins and outs of the rapidly developing high- or superresolution microscopy field, including new methods and data analysis tools that exclusively pertain to these techniques. This integration of membrane biology and advanced imaging techniques emphasizes the interdisciplinary nature of this exciting field. The array of contributions from leading world experts makes this book a valuable tool for the visualization of signaling nanoplatforms by means of cutting-edge optical microscopy tools.
Second-harmonic generation (SHG) microscopy has shown great promise for imaging live cells and tissues, with applications in basic science, medical research, and tissue engineering. Second Harmonic Generation Imaging offers a complete guide to this optical modality, from basic principles, instrumentation, methods, and image analysis to biomedical a
Scientists and engineers have long relied on the power of imaging techniques to help see objects invisible to the naked eye, and thus, to advance scientific knowledge. These experts are constantly pushing the limits of technology in pursuit of chemical imagingâ€"the ability to visualize molecular structures and chemical composition in time and space as actual events unfoldâ€"from the smallest dimension of a biological system to the widest expanse of a distant galaxy. Chemical imaging has a variety of applications for almost every facet of our daily lives, ranging from medical diagnosis and treatment to the study and design of material properties in new products. In addition to highlighting advances in chemical imaging that could have the greatest impact on critical problems in science and technology, Visualizing Chemistry reviews the current state of chemical imaging technology, identifies promising future developments and their applications, and suggests a research and educational agenda to enable breakthrough improvements.
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.