Nanoscopy And Multidimensional Optical Fluorescence Microscopy

"Alberto Diaspro has been choreographing light’s dance for over 20 years, and in Nanoscopy and Multidimensional Optical Fluorescence Microscopy, he has assembled a diverse group of experts to explain the methods they use to coax light to reveal biology’s secrets." — From the Foreword by Daniel Evanko, editor, Nature Methods Nanoscopy and Multidimensional Optical Fluorescence Microscopy demonstrates that the boundaries between sciences do blur at the bottom, especially those that might separate the optical work of physicists and the cellular work of microbiologists. In 18 chapters written by pioneering researchers, this work offers the first comprehensive and current documentation of the cutting-edge research being accomplished in a wide range of photonic devices with revolutionary application. The highlight of the book is its coverage of optical nanoscopy and super-resolution microscopy. The rapid advances in this area over the past few years offer researchers in both photonics and molecular biologya wealth of accomplishment upon which they can build. Offering a complete treatment of this emerging field, this volume: Describes how scientists have exploited the properties of light and its fluorophore partners to overcome the resolution limit of conventional light microscopy Delves into recent ways to minimize the photobleaching that has long hampered many methods including those that have the potential to capture previously unobtainable information on the movements of single molecules Discusses the principles, benefits, and implementation of fluorescence correlation spectroscopy and related methods, which simplifies analysis by limiting light to stationary focal points in a sample Considers the most basic as well as emerging methods for improving three-dimensional optical sectioning microscopy Reviews the basics of FRET (fluorescence resonance energy transfer) and considers its new use for investigating protein complexes The text also introduces those emerging nonfluorescence microscopy methods that can actually exert mechanical forces to trap and move a variety of objects ranging from beads to living cells and cellular organelles. Combining this technique with fluorescence microscopy provides an unparalleled ability to manipulate and visualize biological samples. In the half-century since Richard Feynman challenged scientists to come up with the tools to investigate and manipulate our world at the nanoscale, we have succeeded in placing tools in the hands of biophysicists that are leading to major breakthroughs in our understanding of life and our ability to diagnose, treat, and prevent many challenges to human health. This book reflects what has been accomplished to date while pointing the way to what still needs to be done.

This book describes developments in the field of super-resolution fluorescence microscopy or nanoscopy. In 11 chapters, distinguished scientists and leaders in their respective fields describe different nanoscopy approaches, various labeling technologies, and concrete applications. The topics covered include the principles and applications of the most popular nanoscopy techniques STED and (f)PALM/STORM, along with advances brought about by fluorescent proteins and organic dyes optimized for fluorescence nanoscopy. Furthermore, the photophysics of fluorescent labels is addressed, specifically for improving their photoswitching capabilities. Important applications are also discussed, such as the tracking and counting of molecules to determine acting forces in cells, and quantitative cellular imaging, respectively, as well as the mapping of chemical reaction centers at the nano-scale. The 2014 Chemistry Nobel Prize® was awarded for the ground-breaking developments of super-resolved fluorescence microscopy. In this book, which was co-edited by one of the prize winners, readers will find the most recent developments in this field.

Microscopy is at the forefront of multidisciplinary research. It was developed by physicists, made specific by chemists, and applied by biologists and doctors to better understand how the human body works. For this very reason, the field has been revolutionized in past decades. The objective of Optical Nanoscopy and Novel Microscopy Techniques is to choose some of those revolutionary ideas and serve a general audience from broad disciplines to achieve a fundamental understanding of these technologies and to better apply them in their daily research. The book begins with coverage of super-resolution optical microscopy, which discusses targeted modulation such as STED and SIM or localization methods such as PALM. It then discusses novel development of fluorescent probes, such as organic small-molecule probes, fluorescent proteins, and inorganic labels such as quantum dots. Finally, it describes advanced optical microscopy, such as fluorescence lifetime imaging, fiber optic microscopy, scanning ion conductance microscopy, and the joining of optics and acoustics—photoacoustic microscopy. Following each chapter, a detailed list of references is provided. Problems at the end of each chapter are also included.

This book covers various aspects of modern microscopy, with emphasis on multidimensional (three-dimensional and higher) and multimodality microscopy. The topics discussed include multiphoton fluorescent microscopy, confocal microscopy, x-ray microscopy and microtomography, electron microscopy, probe microscopy and multidimensional image processing for microscopy. In addition, there are chapters demonstrating typical microscopical applications, both biological and material.

In the last decade, fluorescence microscopy has evolved from a classical “retrospective” microscopy approach into an advanced imaging technique that allows the observation of cellular activities in living cells with increased resolution and dimensions. A bright new future has arrived as the nano era has placed a whole new array of tools in the hands of biophysicists who are keen to go deeper into the intricacies of how biological systems work. Following an introduction to the complex world of optical microscopy, this book covers topics such as the concept of white confocal, nonlinear optical microscopy, fluctuation spectroscopies, site-specific labeling of proteins in living cells, imaging molecular physiology using nanosensors, measuring molecular dynamics, muscle braking and stem cell differentiation.

Written by leading optical phase microscopy experts, this book is a comprehensive reference to phase microscopy and nanoscopy techniques for biomedical applications, including differential interference contrast (DIC) microscopy, phase contrast microscopy, digital holographic microscopy, optical coherence tomography, tomographic phase microscopy, spectral-domain phase detection, and nanoparticle usage for phase nanoscopy The Editors show biomedical and optical engineers how to use phase microscopy for visualizing unstained specimens, and support the theoretical coverage with applied content and examples on designing systems and interpreting results in bio- and nanoscience applications. Provides a comprehensive overview of the principles and techniques of optical phase microscopy and nanoscopy with biomedical applications. Tips/advice on building systems and working with advanced imaging biomedical techniques, including interpretation of phase images, and techniques for quantitative analysis based on phase microscopy. Interdisciplinary approach that combines optical engineering, nanotechnology, biology and medical aspects of this topic. Each chapter includes practical implementations and worked examples.

A comprehensive introduction to advanced fluorescence microscopy methods and their applications. This is the first title on the topic designed specifically to allow students and researchers with little background in physics to understand both microscopy basics and novel light microscopy techniques. The book is written by renowned experts and pioneers in the field with a rather intuitive than formal approach. It always keeps the nonexpert reader in mind, making even unavoidable complex theoretical concepts readily accessible. All commonly used methods are covered. A companion website with additional references, examples and video material makes this a valuable teaching resource: http://www.wiley-vch.de/home/fluorescence_microscopy/

This book covers various aspects of modern microscopy, with emphasis on multidimensional (three-dimensional and higher) and multimodality microscopy. The topics discussed include multiphoton fluorescent microscopy, confocal microscopy, x-ray microscopy and microtomography, electron microscopy, probe microscopy and multidimensional image processing for microscopy. In addition, there are chapters demonstrating typical microscopical applications, both biological and material. Contents: Fluorescence Lifetime Imaging by Double Pulse Excitation in Bilateral Confocal Microscopy (G J Brakenhoff et al.)Icosahedral Virus Structure Determination by Electron Cryomicroscopy: Image Processing Principles (P A Thuman-Commike & W Chiu)Appropriate Image Processing for Confocal Microscopy (G Cox & C J R Sheppard)X-Ray Fluorescence Two-Dimensional Microanalysis at the VEPP-3 Storage Ring: Applications in Environmental Science (I P Dolbnya & K V Zolotarev)3D Transfer-Function Analysis for 4Pi Confocal Microscopy (M Gu & C J R Sheppard)Image Restoration of Thick Biological Specimens for Transmission Electron Microscope Tomography (K F Han et al.)Images of Membrane-Bound Detoxification Enzyme at 4Å Resolution Obtained by Electron Cryomicroscopy (I Schmidt-Krey et al.)The Uptake Pathway of DNA and Lipids in Cationic Liposome-Mediated Gene Transfer (S-W Hui et al.)Multiscale Imaging, Analysis and Modeling of the Pulmonary Bronchial Tree (A Kriete et al.)Asymmetric Cell Division as a Mechanism of Cell Determination in Vascular Plants (B-L Lin)3D Electron Microscopy of Cells and Organelles by High-Voltage EM, Tomography and Stereopair Analysis (M Marko)Surface Scattering Techniques in Scanning Optical Microscopy (J F Aguilar & E R Méndez)Applications of Electron Microscopy to Materials and Earth Sciences – Dislocation, Shape and Orientation Changes and Interdiffusion (P Shen)Surface Profiling and Confocal Microscopy (C J R Sheppard & J C Quartel)Matched Spatial Filters in Long Working Distance Microscopy of Phase Objects (J R Strickler & J-S Hwang)Transport Pathways of Lipoproteins Across the Arterial Endothelial Cells (C-H Kao et al.)Polarisation Effects in Confocal Microscopy (T Wilson)Near-Field Microscopy and Spectroscopy of Single Molecules, Single Proteins and Biological Membranes (X S Xie et al.) Keywords:Microscopy;Optics;Image Processing