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.

Optical microscopy is one of the most frequently used tools in chemistry and the life sciences. However, its limited resolution hampers the use of optical imaging to many other relevant problems in different disciplines. Super-Resolution Microscopy (SRM) is a new technique that allows the resolution of objects down to a few billionth of meters (nanometers), ten times better than classical microscopes, opening up opportunities to use this tool in new fields. This book describes the theory, principles, and practice of super-resolution microscopy in the field of materials science and nanotechnology. There is a growing interest in the applications of SRM beyond biology as new synthetic materials, such as nanoscale sensors and catalysts, nanostructured materials, functional polymers, and nanoparticles, have nanoscopic features that are challenging to visualize with traditional imaging methods. SRM has the potential to be used to image and understand these cutting-edge man-made objects and guide the design of materials for novel applications. This book is an ideal guide for researchers in the fields of microscopy and materials science and chemistry as well as graduate students studying physics, materials science, biomedical engineering, and chemistry. Key Features: Contains practical guidance on Super-Resolution Microscopy (SRM), an exciting and growing tool that was awarded the Nobel Prize for chemistry in 2014 Provides a new perspective targeting materials science, unlike existing books which target readers in chemistry, life science, and biology Targets students in its core chapters, while offering more advanced material for professionals and researchers in later chapters

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.

Super Resolution Optical Imaging and Microscopy Extremely comprehensive resource containing cutting-edge and practical knowledge of super-resolution optical imaging This book covers both the basic principles and specific technical details of super-resolution microscopy techniques. It covers the criteria to choose different fluorophores for various SRM methods and critically assesses the nitty-gritty of associated problems that are often encountered in practical applications. A progressive guide to designing the next generation of advanced fluorophores to meet the goal of advanced SR imaging studies is also put forward. Written by two well-qualified authors, the book contains exclusive content to enhance readers’ understanding on innovation of newer SRM technologies. Sample topics covered in the book include: Optical techniques, fluorescent probe design, and algorithm development Recent highlight and breakthroughs in biology using SRM methods The overall success of SRM in biological inventions The future direction and scope of the field This book is an invaluable resource for chemists and researchers/scientists involved in designing newer fluorescent materials for SRM studies. It can also assist biologists engaged in advanced biological studies using SRM by guiding them through sample preparation, image processing, and precautions to be taken in practical imaging studies.

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.

Plasmonics is an emerging field mainly developed within the past two decades. Due to its unique capabilities to manipulate light at deep subwavelength scales, plasmonics has been commonly treated as the most important part of nanophotonics. Plasmonic-assisted optical microscopy techniques, especially super-resolution microscopy, have shown tremendous potential and attracted much attention. This book aims to collect cutting-edge studies in various optical imaging technologies with advanced performances that are enabled or enhanced by plasmonics. The basic working principles, development details, and potential future direction and perspectives are discussed. Edited by Zhaowei Liu, a prominent researcher in the field of super-resolution microscopy, this book will be an excellent reference for anyone in the field of nanophotonics, plasmonics, and optical microscopy.

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.

A comprehensive guide to the art and science of bioimaging data acquisition, processing and analysis Standard and Super-Resolution Bioimaging Data Analysis gets newcomers to bioimage data analysis quickly up to speed on the mathematics, statistics, computing hardware and acquisition technologies required to correctly process and document data. The past quarter century has seen remarkable progress in the field of light microscopy for biomedical science, with new imaging technologies coming on the market at an almost annual basis. Most of the data generated by these systems is image-based, and there is a significant increase in the content and throughput of these imaging systems. This, in turn, has resulted in a shift in the literature on biomedical research from descriptive to highly-quantitative. Standard and Super-Resolution Bioimaging Data Analysis satisfies the demand among students and research scientists for introductory guides to the tools for parsing and processing image data. Extremely well illustrated and including numerous examples, it clearly and accessibly explains what image data is and how to process and document it, as well as the current resources and standards in the field. A comprehensive guide to the tools for parsing and processing image data and the resources and industry standards for the biological and biomedical sciences Takes a practical approach to image analysis to assist scientists in ensuring scientific data are robust and reliable Covers fundamental principles in such a way as to give beginners a sound scientific base upon which to build Ideally suited for advanced students having only limited knowledge of the mathematics, statistics and computing required for image data analysis An entry-level text written for students and practitioners in the bioscience community, Standard and Super-Resolution Bioimaging Data Analysis de-mythologises the vast array of image analysis modalities which have come online over the past decade while schooling beginners in bioimaging principles, mathematics, technologies and standards.

Introduces readers to the enlightening world of the modern light microscope There have been rapid advances in science and technology over the last decade, and the light microscope, together with the information that it gives about the image, has changed too. Yet the fundamental principles of setting up and using a microscope rests upon unchanging physical principles that have been understood for years. This informative, practical, full-colour guide fills the gap between specialised edited texts on detailed research topics, and introductory books, which concentrate on an optical approach to the light microscope. It also provides comprehensive coverage of confocal microscopy, which has revolutionised light microscopy over the last few decades. Written to help the reader understand, set up, and use the often very expensive and complex modern research light microscope properly, Understanding Light Microscopy keeps mathematical formulae to a minimum—containing and explaining them within boxes in the text. Chapters provide in-depth coverage of basic microscope optics and design; ergonomics; illumination; diffraction and image formation; reflected-light, polarised-light, and fluorescence microscopy; deconvolution; TIRF microscopy; FRAP & FRET; super-resolution techniques; biological and materials specimen preparation; and more. Gives a didactic introduction to the light microscope Encourages readers to use advanced fluorescence and confocal microscopes within a research institute or core microscopy facility Features full-colour illustrations and workable practical protocols Understanding Light Microscopy is intended for any scientist who wishes to understand and use a modern light microscope. It is also ideal as supporting material for a formal taught course, or for individual students to learn the key aspects of light microscopy through their own study.