Label Free Super Resolution Microscopy

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.

Fluorescence Microscopy: Super-Resolution and other Novel Techniques delivers a comprehensive review of current advances in fluorescence microscopy methods as applied to biological and biomedical science. With contributions selected for clarity, utility, and reproducibility, the work provides practical tools for investigating these ground-breaking developments. Emphasizing super-resolution techniques, light sheet microscopy, sample preparation, new labels, and analysis techniques, this work keeps pace with the innovative technical advances that are increasingly vital to biological and biomedical researchers. With its extensive graphics, inter-method comparisons, and tricks and approaches not revealed in primary publications, Fluorescence Microscopy encourages readers to both understand these methods, and to adapt them to other systems. It also offers instruction on the best visualization to derive quantitative information about cell biological structure and function, delivering crucial guidance on best practices in related laboratory research. Presents a timely and comprehensive review of novel techniques in fluorescence imaging as applied to biological and biomedical research Offers insight into common challenges in implementing techniques, as well as effective solutions

Stimulated Raman Scattering Microscopy: Techniques and Applications describes innovations in instrumentation, data science, chemical probe development, and various applications enabled by a state-of-the-art stimulated Raman scattering (SRS) microscope. Beginning by introducing the history of SRS, this book is composed of seven parts in depth including instrumentation strategies that have pushed the physical limits of SRS microscopy, vibrational probes (which increased the SRS imaging functionality), data science methods, and recent efforts in miniaturization. This rapidly growing field needs a comprehensive resource that brings together the current knowledge on the topic, and this book does just that. Researchers who need to know the requirements for all aspects of the instrumentation as well as the requirements of different imaging applications (such as different types of biological tissue) will benefit enormously from the examples of successful demonstrations of SRS imaging in the book. Led by Editor-in-Chief Ji-Xin Cheng, a pioneer in coherent Raman scattering microscopy, the editorial team has brought together various experts on each aspect of SRS imaging from around the world to provide an authoritative guide to this increasingly important imaging technique. This book is a comprehensive reference for researchers, faculty, postdoctoral researchers, and engineers. Includes every aspect from theoretic reviews of SRS spectroscopy to innovations in instrumentation and current applications of SRS microscopy Provides copious visual elements that illustrate key information, such as SRS images of various biological samples and instrument diagrams and schematics Edited by leading experts of SRS microscopy, with each chapter written by experts in their given topics

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 Imaging and Sensing Understand the future of optical imaging with this cutting-edge guide Optoelectronic devices for imaging and sensing are among the backbones of modern technology. Facilitating the mutual conversion of optical and electrical signals, they have applications from telecommunications to molecular spectroscopy, and their incorporation into photon-involved technologies is only growing. The rapid development of this field makes the need for a fully up-to-date introduction all the more critical. Optical Imaging and Sensing meets this need with a comprehensive guide to the novel materials and devices employed in optical imaging and sensing. Given the current revolution in new imaging materials, an introduction that fully incorporates the latest research is an indispensable tool for scientists and engineers in a huge range of fields. The technologies surveyed here promise to transform public security, 5G and next-generation wireless communication, clinical imaging, and many more. Optical Imaging and Sensing Readers will also find: Detailed discussion of materials including semimetallic graphene, semiconducting black phosphorous, and many more Discussion of devices from infrared photodetectors to nonlinear interferometers A thorough look forward to the future of the field Optical Imaging and Sensing is a useful reference for materials scientists, spectroscopists, semiconductor physicists, and engineers working in any field or industry involving optical imaging or sensing technology.

Optical microscopy is developing into nanoscopy and multimodal microscopy the better to decipher the functioning mechanisms in living systems, and investigating biological specimens at molecular level using fluorescence as a mechanism of contrast. Results have demonstrated the potential to provide information at the Angstrom level. Other optical methods now offer more in terms of spatial and temporal resolution, making it possible to study the delicate and complex relationship between structure and function in cells. Modern optical microscopes also use the decisive advantage provided by artificial intelligence algorithms. All in all, a rapidly changing field with an increasing number of questions to be answered. This book contains the lectures presented as Course 210 of the prestigious International School of Physics Enrico Fermi, Multimodal and Nanoscale Optical Microscopy, held in Varenna, Italy from 11 to 16 July 2021. Topics covered include fluorescence; linear and nonlinear microscopy; label-free with Mueller matrix and Brillouin microscopy; F-methods such as FRAP, FLIM, FRET and FCS; super-resolution, phototoxicity and photodamage; optical and magnetic trapping; image formation; and bioimage analysis, among others. The book also includes selected contributions of a number of young researchers. Packed with novel ideas, new instruments, challenging theoretical approaches and amazing applications, the book will be of interest to all those working in the field of multimodal and nanoscale optical microscopy.

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 is dedicated to label-free, non-invasive monitoring of cell-based assays and it comprises the most widely applied techniques. Each approach is described and critically evaluated by an expert in the field such that researchers get an overview on what is possible and where the limitations are. The book provides the theoretical basis for each technique as well as the most successful and exciting applications. Label-free bioanalytical techniques have been known for a long time as valuable tools to monitor adsorption processes at the solid-liquid interface in general – and biomolecular interaction analysis (BIA) in particular. The underlying concepts have been progressively transferred to the analysis of cell-based assays. The strength of these approaches is implicitly given with the name 'label-free': the readout is independent of any label, reagent or additive that contaminates the system under study and potentially affects its properties. Thus, label-free techniques provide an unbiased analytical perspective in the sense that the sample is not manipulated by additives but pure. They are commonly based on physical principles and read changes in integral physical properties of the sample like refractive index, conductivity, capacitance or elastic modulus to mention just a few. Even though it is not implied in the name, label-free approaches usually monitor the cells under study non-invasively meaning that the amplitude of the signal (e.g. electric field strength, mechanical elongation) that is used for the measurement is too low to interfere or affect. In contrast to label-based analytical techniques that are commonly restricted to a single reading at a predefined time point, label-free approaches allow for a continuous observation so that the dynamics of the biological system or reaction become accessible.