· 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

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.

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.

New Methods and Sensors for Membrane and Cell Volume Research, Volume 88 provides an overview of novel experimental approaches to study both the cell membrane and the under-membrane space – the cytosol, which have lately began drawing renewed attention. The book's overall emphasis is on fluorescent and FRET-based sensors, however, other optical (such as variants of transmission microscopy) and non-optical methods (neutron scattering and mass spectrometry) also have dedicated chapters. This volume provides a rare review of experimental approaches to study intracellular phase transitions, as well as anion channels, membrane tension and dynamics, and other topics of intense current interest. - Describes novel FRET-based membrane sensors - Reviews selected non-optical approaches to membrane structure and dynamics - Describes traditional and modern aspects of cell volume research, such as phase transitions and macromolecular crowding

Biological membranes play a central role in cell structure, shape and functions. However, investigating the membrane bilayer has proved to be difficult due to its highly dynamic and anisotropic structure, which generates steep gradients at the nanometer scale. Due to the decisive impact of recently developed fluorescence-based techniques, tremendous advances have been made in the last few years in our understanding of membrane characteristics and functions. In this context, the present book illustrates some of these major advances by collecting review articles written by highly respected experts. The book is organized in three parts, the first of which deals with membrane probes and model membranes. The second part describes the use of advanced quantitative and high-resolution techniques to explore the properties of biological membranes, illustrating the key progress made regarding membrane organization, dynamics and interactions. The third part is focused on the investigation of membrane proteins using the same techniques, and notably on the membrane receptors that play a central role in signaling pathways and therapeutic strategies. All chapters provide comprehensive information on membranes and their exploration for beginners in the field and advanced researchers alike.

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.

Current Topics in Membranes is targeted toward scientists and researchers in biochemistry and molecular and cellular biology, providing the necessary membrane research to assist them in discovering the current state of a particular field and in learning where that field is heading. This volume offers an up to date presentation of current knowledge in the field of Lipid Domains. - Written by leading experts - Contains original material, both textual and illustrative, that should become a very relevant reference material - The material is presented in a very comprehensive manner - Both researchers in the field and general readers should find relevant and up-to-date information

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.

This book highlights recent advances in and diverse techniques for exploring the plasma membrane’s structure and function. It starts with two chapters reviewing the history of membrane research and listing recent advances regarding membrane structure, such as the semi-mosaic model for red blood cell membranes and the protein layer-lipid-protein island model for nucleated tissue cell membranes. It subsequently focuses on the localization and interactions of membrane components, dynamic processes of membrane transport and transmembrane signal transduction. Classic and cutting-edge techniques (e.g. high-resolution atomic force microscopy and super-resolution fluorescence microscopy) used in biophysics and chemistry are presented in a very comprehensive manner, making them useful and accessible to both researchers in the field and novices studying cell membranes. This book provides readers a deeper understanding of the plasma membrane’s organization at the single molecule level and opens a new way to reveal the relationship between the membrane’s structure and functions, making it essential reading for researchers in various fields.