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This book presents the advances in super-resolution microscopy in physics and biomedical optics for nanoscale imaging. In the last decade, super-resolved fluorescence imaging has opened new horizons in improving the resolution of optical microscopes far beyond the classical diffraction limit, leading to the Nobel Prize in Chemistry in 2014. This book represents the first comprehensive review of a different type of super-resolved microscopy, which does not rely on using fluorescent markers. Such label-free super-resolution microscopy enables potentially even broader applications in life sciences and nanoscale imaging, but is much more challenging and it is based on different physical concepts and approaches. A unique feature of this book is that it combines insights into mechanisms of label-free super-resolution with a vast range of applications from fast imaging of living cells to inorganic nanostructures. This book can be used by researchers in biological and medical physics. Due to its logically organizational structure, it can be also used as a teaching tool in graduate and upper-division undergraduate-level courses devoted to super-resolved microscopy, nanoscale imaging, microscopy instrumentation, and biomedical imaging.
This book presents a comprehensive and coherent summary of techniques for enhancing the resolution and image contrast provided by far-field optical microscopes. It takes a critical look at the body of knowledge that comprises optical microscopy, compares and contrasts the various instruments, provides a clear discussion of the physical principles that underpin these techniques, and describes advances in science and medicine for which superresolution microscopes are required and are making major contributions. The text fills significant gaps that exist in other works on superresolution imaging, firstly by placing a new emphasis on the specimen, a critical component of the microscope setup, giving equal importance to the enhancement of both resolution and contrast. Secondly, it covers several topics not typically discussed in depth, such as Bessel and Airy beams, the physics of the spiral phase plate, vortex beams and singular optics, photoactivated localization microscopy (PALM), stochastic optical reconstruction microscopy (STORM), structured illumination microscopy (SIM), and light-sheet fluorescence microscopy (LSFM). Several variants of these techniques are critically discussed. Noise, optical aberrations, specimen damage, and artifacts in microscopy are also covered. The importance of validation of superresolution images with electron microscopy is stressed. Additionally, the book includes translations and discussion of seminal papers by Abbe and Helmholtz that proved to be pedagogically relevant as well as historically significant. This book is written for students, researchers, and engineers in the life sciences, medicine, biological engineering, and materials science who plan to work with or already are working with superresolution light microscopes. The volume can serve as a reference for these areas while a selected set of individual chapters can be used as a textbook for a one-semester undergraduate or first-year graduate course on superresolution microscopy. Moreover, the text provides a captivating account of curiosity, skepticism, risk-taking, innovation, and creativity in science and technology. Good scientific practice is emphasized throughout, and the author’s lecture slides on responsible conduct of research are included as an online resource which will be of interest to students, course instructors, and scientists alike.
This open access book provides a comprehensive overview of the application of the newest laser and microscope/ophthalmoscope technology in the field of high resolution imaging in microscopy and ophthalmology. Starting by describing High-Resolution 3D Light Microscopy with STED and RESOLFT, the book goes on to cover retinal and anterior segment imaging and image-guided treatment and also discusses the development of adaptive optics in vision science and ophthalmology. Using an interdisciplinary approach, the reader will learn about the latest developments and most up to date technology in the field and how these translate to a medical setting. High Resolution Imaging in Microscopy and Ophthalmology – New Frontiers in Biomedical Optics has been written by leading experts in the field and offers insights on engineering, biology, and medicine, thus being a valuable addition for scientists, engineers, and clinicians with technical and medical interest who would like to understand the equipment, the applications and the medical/biological background. Lastly, this book is dedicated to the memory of Dr. Gerhard Zinser, co-founder of Heidelberg Engineering GmbH, a scientist, a husband, a brother, a colleague, and a friend.
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.
This book encompasses the full breadth of the super-resolution imaging field, representing modern techniques that exceed the traditional diffraction limit, thereby opening up new applications in biomedicine. It shows readers how to use the new tools to increase resolution in sub-nanometer-scale images of living cells and tissue, which leads to new information about molecules, pathways and dynamics. The book highlights the advantages and disadvantages of the techniques, and gives state-of-the-art examples of applications using microscopes currently available on the market. It covers key techniques such as stimulated emission depletion (STED), structured illumination microscopy (SSIM), photoactivated localization microscopy (PALM), and stochastic optical reconstruction microscopy (STORM). It will be a useful reference for biomedical researchers who want to work with super-resolution imaging, learn the proper technique for their application, and simultaneously obtain a solid footing in other techniques.
This book describes how to see atoms using electron microscopes. This new edition includes updated sections on applications and new uses of atomic-resolution transmission electron microscopy. Several new chapters and sources of software for image interpretation and electron-optical design have also been added.
Expansion Microscopy for Cell Biology, Volume 161 in the Methods in Cell Biology series, compiles recent developments in expansion microscopy techniques (Pro-ExM, U-ExM, Ex-STED, X10, Ex-dSTORM, etc.) and their applications in cell biology, ranging from mitosis, centrioles or nuclear pore complex to plant cell, bacteria, Drosophila or neurons. Chapters in this new release include Protein-retention Expansion Microscopy: Improved Sub-cellular Imaging Resolution through Physical Specimen Expansion, Ultrastructure Expansion Microscopy (U-ExM), Expansion STED microscopy (ExSTED), Simple multi-color super-resolution by X10 microscopy, Expansion microscopy imaging of various neuronal structures, Mapping the neuronal cytoskeleton using expansion microscopy, Mechanical expansion microscopy, and much more. - Provides the authority and expertise of leading contributors from an international board of authors - Represents the latest release in the Methods in Cell Biology series - Includes the latest information on Expansion Microscopy for Cell Biology
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.
The topics range from single molecule experiments in quantum optics and solid-state physics to analogous investigations in physical chemistry and biophysics.
This unique book on super-resolution microscopy techniques presents comparative, in-depth analyses of the strengths and weaknesses of the individual approaches. It was written for non-experts who need to understand the principles of super-resolution or who wish to use recently commercialized instruments as well as for professionals who plan to realize novel microscopic devices. Explaining the practical requirements in terms of hardware, software and sample preparation, the book offers a wealth of hands-on tips and practical tricks to get a setup running, provides invaluable help and support for successful data acquisition and specific advice in the context of data analysis and visualization. Furthermore, it addresses a wide array of transdisciplinary fields of applications. The author begins by outlining the joint efforts that have led to achieving super-resolution microscopy combining advances in single-molecule photo-physics, fluorophore design and fluorescent labeling, instrument design and software development. The following chapters depict and compare current main standard techniques such as structured illumination microscopy, single-molecule localization, stimulated emission depletion microscopy and multi-scale imaging including light-sheet and expansion microscopy. For each individual approach the experimental setups are introduced, the imaging protocols are provided and the various applications illustrated. The book concludes with a discussion of future challenges addressing issues of routine applications and further commercialization of the available methods. Guiding users in how to make choices for the design of their own experiments from scratch to promising application, this one-stop resource is intended for researchers in the applied sciences, from chemistry to biology and medicine to physics and engineering.