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 (R) 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.

Confocal Microscopy: Methods and Protocols, Second Edition takes the researcher from the bench top through the imaging process, to the page. Protocols for the preparation of tissues from many model organisms including worms, flies and mice have been included as well as chapters on confocal imaging of living cells, three dimensional analysis, and the measurement and presentation of confocal images for publication. Emphasis has been placed on the laser scanning confocal microscope since this is still the instrument used for most routine applications. The current generation of modern confocal instruments produces optical sections of cells and tissues that are free of out-of-focus fluorescence with reduced chances of artifacts from the techniques of specimen preparation. This allows the imaging of living specimens and measurements of physiological events within cells. Confocal microscopy has become essential in many fields of contemporary biomedical research where a light microscope is required for imaging fluorescently labeled cells and tissues, especially cell biology, developmental biology, neurobiology, and pathology. Written in the successful Methods in Molecular Biology series format, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible protocols, and notes on troubleshooting and avoiding known pitfalls. Authoritative and easily accessible, Confocal Microscopy: Methods and Protocols, Second Edition is aimed primarily, but not exclusively, at the novice user with pointers to more advanced techniques.

Microscope Image Processing, Second Edition, introduces the basic fundamentals of image formation in microscopy including the importance of image digitization and display, which are key to quality visualization. Image processing and analysis are discussed in detail to provide readers with the tools necessary to improve the visual quality of images, and to extract quantitative information. Basic techniques such as image enhancement, filtering, segmentation, object measurement, and pattern recognition cover concepts integral to image processing. In addition, chapters on specific modern microscopy techniques such as fluorescence imaging, multispectral imaging, three-dimensional imaging and time-lapse imaging, introduce these key areas with emphasis on the differences among the various techniques. The new edition discusses recent developments in microscopy such as light sheet microscopy, digital microscopy, whole slide imaging, and the use of deep learning techniques for image segmentation and analysis with big data image informatics and management. Microscope Image Processing, Second Edition, is suitable for engineers, scientists, clinicians, post-graduate fellows and graduate students working in bioengineering, biomedical engineering, biology, medicine, chemistry, pharmacology and related fields, who use microscopes in their work and would like to understand the methodologies and capabilities of the latest digital image processing techniques or desire to develop their own image processing algorithms and software for specific applications.

A comprehensive tutorial for researchers and practitioners involved in surface science. The basics of the scanning probe microscopy techniques as well as material class-specific applications are thoroughly discussed. The book gives access to these methods for advanced students and allows researchers to apply these powerful atomic-resolution imaging techniques to new systems.

This book represents the compilation of papers presented at the second Atomic Force Microscopy/Scanning Tunneling Microscopy (AFM/STM) Symposium, held June 7 to 9, 1994, in Natick, Massachusetts, at Natick Research, Development and Engineering Center, now part ofU.S. Army Soldier Systems Command. As with the 1993 symposium, the 1994 symposium provided a forum where scientists with a common interest in AFM, STM, and other probe microscopies could interact with one another, exchange ideas and explore the possibilities for future collaborations and working relationships. In addition to the scheduled talks and poster sessions, there was an equipment exhibit featuring the newest state-of-the-art AFM/STM microscopes, other probe microscopes, imaging hardware and software, as well as the latest microscope-related and sample preparation accessories. These were all very favorably received by the meeting's attendees. Following opening remarks by Natick's Commander, Colonel Morris E. Price, Jr., and the Technical Director, Dr. Robert W. Lewis, the symposium began with the Keynote Address given by Dr. Michael F. Crommie from Boston University. The agenda was divided into four major sessions. The papers (and posters) presented at the symposium represented a broad spectrum of topics in atomic force microscopy, scanning tunneling microscopy, and other probe microscopies.

This comprehensive reference illustrates optimal preparation methods in biological electron microscopy compared with common methodological problems. Not only will the basic methodologies of transmission electron microscopy like fixation, microtomy, and microscopy be presented, but the authors also endeavor to illustrate more specialized techniques such as negative staining, autoradiography, cytochemistry, immunoelectron microscopy, and computer-assisted image analysis.
Key Features
* Authored by the key leaders in the biological electron microscopy field
* Illustrates both optimal and suboptimal or artifactual results in a variety of electron microscopy disciplines
* Introduces students on how to read and interpret electron micrographs

Aquaculture is rapidly becoming a major source of fish protein used to meet the nutritional needs of humans. As the aquaculture industry grows, exposure of farmed fish to environmental contaminants, and the need for chemical therapeutic agents for fish, will increase. This book is designed to bring together authorities worldwide on the regulation of environmental contaminants and food chemicals and researchers investigating the metabolism and disposition of foreign chemicals (xenobiotics) in fish species.

In modern scanning electron microscopy, sample surface preparation is of key importance, just as it is in transmission electron microscopy. With the procedures for sample surface preparation provided in the present book, the enormous potential of advanced scanning electron microscopes can be realized fully. This will take the reader to an entirely new level of scanning electron microscopy and finely-detailed images never seen before. Written for: Scientists, practitioners, academic libraries, graduate students

Transmission electron microscopy (TEM) is now recognized as a crucial tool in materials science. This book, authored by a team of expert Chinese and international authors, covers many aspects of modern electron microscopy, from the architecture of novel electron microscopes, advanced theories and techniques in TEM and sample preparation, to a variety of hands-on examples of TEM applications. Volume I concentrates on the newly developed concepts and methods which are making TEM a powerful and indispensible tool in materials science.

Friction force microscopy is an important analytical tool in the field of tribology on the nanometer-scale. The contact area between the probing tip and the sample is reduced to some square nanometers, corresponding to the ideal of a single asperity contact. Traditional concepts, such as friction coefficients, adhesion and elasticity and stick-slip are re-examined with this novel technique. New concepts based upon classical and quantum mechanics are investigated.

Transmission electron microscopy (TEM) is now recognized as a crucial tool in materials science. This book, authored by a team of expert Chinese and international authors, covers many aspects of modern electron microscopy, from the architecture of novel electron microscopes, advanced theories and techniques in TEM and sample preparation, to a variety of hands-on examples of TEM applications. Volume II illustrates the important role that TEM is playing in the development and characterization of advanced materials, including nanostructures, interfacial structures, defects, and macromolecular complexes.

This book discusses the various principles in confocal scanning microscopy which has become a useful tool in many practical fields including biological studies and industrial inspection. The methodology presented in this book is unique and is based on the concept of the three-dimensional transfer functions which have been developed by the author and his colleagues over the last five years. With the 3-D transfer functions, resolving power in 3-D confocal imaging can be defined in a unified way, different optical arrangements can be compared with an insight into their inter-relationship, and images of thick objects can be modeled in terms of the Fourier transform which makes the analysis easy. The aim of this book is to provide a systematic introduction to the concept of the 3-D transfer functions in various confocal microscopes, to describe the methods for the derivation of different 3-D transfer functions, and to explain the principles of 3-D confocal imaging in terms of these functions.

Fluorescence microscopy images can be easily integrated into current video and computer image processing systems. People like visual observation; they like to watch a television or computer screen, and fluorescence techniques are thus becoming more and more popular. Since true in vivo experiments are simple to perform, samples can be directly seen and there is always the possibility of manipulating the samples during the experiments; it is an ideal technique for biology and medicine. Images are obtained by a classical (now called wide-field) fluorescence microscope, a confocal scanning microscope, upright or inverted, with epifluorescence or transmission. Computerized image processing may improve definition, and remove glare and scattered light signal. It also makes it possible to compute ratio images (ratio imaging both in excitation and in emission) or lifetime imaging. Image analysis programs may supply a great deal of additional data of various types, starting with calculations of the number of fluorescent objects, their shapes, brightness, etc. Fluorescence microscopy data may be complemented by classical measurement in the cuvette yr by flow cytometry.

2. High Temperature UHV-STM System 264 3. Hydrogen Desorption Process on Si (111) Surface 264 4. (7x7) - (1 xl) Phase Transition on Si (111) Surface 271 Step Shifting under dc Electric Fields 275 5. 6. Conclusions 280 Acknowledgements and References 281 12. DYNAMIC OBSERVATION OF VORTICES IN SUPERCONDUCTORS USING ELECTRON WAVES 283 by Akira Tonomura 1. Introduction 283 2. Experimental Method 284 2. 1 Interference Microscopy 284 2. 2 Lorentz Microscopy 287 Observation of Superconducting Vortices 288 3. 3. 1 Superconducting Vortices Observed by Interference Microscopy 288 3. 1. 1 Profile Mode 288 3. 1. 2 Transmission Mode 291 3. 2 Superconducting Vortices Observed by Lorentz Microscopy 293 3. 3 Observation of Vortex Interaction with Pinning Centers 294 3. 3. 1 Surface Steps 295 3. 3. 2 Irradiated Point Defects 296 4. Conclusion 298 References 299 13. TEM STUDIES OF SOME STRUCTURALLY FLEXIBLE SOLIDS AND THEIR ASSOCIATED PHASE TRANSFORMATIONS 301 by Ray L. Withers and John G. Thompson 1. Introduction 301 2. Tetrahedrally Comer-Connected Framework Structures 302 3. Tetragonal a-PbO 311 4. Compositionally Flexible Anion-Deficient Fluorites and the "Defect Fluorite" to C-type Sesquioxide Transition 320 5. Summary and Conclusions 327 Acknowledgements and References 327 Author Index 331 Subject Index 333 List of Contributors A. ASEEV Institute of Semiconductor Physics, Russian Academy of Sciences Novosibirsk, 630090, pr. ac. , Lavrentjeva 13, RUSSIA E. BAUER Department of Physics and Astronomy, Arizona State University Tempe, AZ 85287-1504, U. S. A. G. H.

The first U. S. Army Natick Research, Development and Engineering Center Atomic Force/Scanning Tunneling Microscopy (AFM/STM) Symposium was held on lune 8-10, 1993 in Natick, Massachusetts. This book represents the compilation of the papers presented at the meeting. The purpose ofthis symposium was to provide a forum where scientists from a number of diverse fields could interact with one another and exchange ideas. The various topics inc1uded application of AFM/STM in material sciences, polymers, physics, biology and biotechnology, along with recent developments inc1uding new probe microscopies and frontiers in this exciting area. The meeting's format was designed to encourage communication between members of the general scientific community and those individuals who are at the cutting edge of AFM, STM and other probe microscopies. It immediately became clear that this conference enabled interdisciplinary interactions among researchers from academia, industry and government, and set the tone for future collaborations. Expert scientists from diverse scientific areas including physics, chemistry, biology, materials science and electronics were invited to participate in the symposium. The agenda of the meeting was divided into three major sessions. In the first session, Biological Nanostructure, topics ranged from AFM ofDNA to STM imagmg ofthe biomoleeule tubulin and bacterialluciferase to the AFM of starch polymer double helices to AFM imaging of food surfaces.

This thesis reports on the development of the first quantum enhanced microscope and on its applications in biological microscopy. The first quantum particle-tracking microscope, described in detail here, represents a pioneering advance in quantum microscopy, which is shown to be a powerful and relevant technique for future applications in science and medicine. The microscope is used to perform the first quantum-enhanced biological measurements -- a central and long-standing goal in the field of quantum measurement. Sub diffraction-limited quantum imaging is achieved, also for the first time, with a scanning probe imaging configuration allowing 10-nanometer resolution.

Following three printings of the First Edition (1978), the publisher has asked for a Second Edition to bring the contents up to date. In doing so the authors aim to show how the newer microscopies are related to the older types with respect to theoretical resolving power (what you pay for) and resolution (what you get). The book is an introduction to students, technicians, technologists, and scientists in biology, medicine, science, and engineering. It should be useful in academic and industrial research, consulting, and forensics; how ever, the book is not intended to be encyclopedic. The authors are greatly indebted to the College of Textiles of North Carolina State University at Raleigh for support from the administration there for typing, word processing, stationery, mailing, drafting diagrams, and general assistance. We personally thank Joann Fish for word process ing, Teresa M. Langley and Grace Parnell for typing services, Mark Bowen for drawing graphs and diagrams, Chuck Gardner for photographic ser vices, Deepak Bhattavahalli for his work with the proofs, and all the other people who have given us their assistance. The authors wish to acknowledge the many valuable suggestions given by Eugene G. Rochow and the significant editorial contributions made by Elizabeth Cook Rochow."

Bringing Scanning Probe Microscopy Up to Speed introduces the principles of scanning probe systems with particular emphasis on techniques for increasing speed. The authors include useful information on the characteristics and limitations of current state-of-the-art machines as well as the properties of the systems that will follow in the future. The basic approach is two-fold. First, fast scanning systems for single probes are treated and, second, systems with multiple probes operating in parallel are presented. The key components of the SPM are the mechanical microcantilever with integrated tip and the systems used to measure its deflection. In essence, the entire apparatus is devoted to moving the tip over a surface with a well-controlled force. The mechanical response of the actuator that governs the force is of the utmost importance since it determines the scanning speed. The mechanical response relates directly to the size of the actuator; smaller is faster. Traditional scanning probe microscopes rely on piezoelectric tubes of centimeter size to move the probe. In future scanning probe systems, the large actuators will be replaced with cantilevers where the actuators are integrated on the beam. These will be combined in arrays of multiple cantilevers with MEMS as the key technology for the fabrication process.

Since the publication in 1979 of Introduction to Analytical Electron Microscopy (ed. J. J. Hren, J. I. Goldstein, and D. C. Joy; Plenum Press), analytical electron microscopy has continued to evolve and mature both as a topic for fundamental scientific investigation and as a tool for inorganic and organic materials characterization. Significant strides have been made in our understanding of image formation, electron diffraction, and beam/specimen interactions, both in terms of the "physics of the processes" and their practical implementation in modern instruments. It is the intent of the editors and authors of the current text, Principles of Analytical Electron Microscopy, to bring together, in one concise and readily accessible volume, these recent advances in the subject. The text begins with a thorough discussion of fundamentals to lay a foundation for today's state-of-the-art microscopy. All currently important areas in analytical electron microscopy-including electron optics, electron beam/specimen interactions, image formation, x-ray microanalysis, energy-loss spectroscopy, electron diffraction and specimen effects-have been given thorough attention. To increase the utility of the volume to a broader cross section of the scientific community, the book's approach is, in general, more descriptive than mathematical. In some areas, however, mathematical concepts are dealt with in depth, increasing the appeal to those seeking a more rigorous treatment of the subject.

This monograph stems from the lectures given during the summer course at the University of La Laguna, Canary Islands, Spain. It includes the main characterization techniques useful nowadays for ceramics, glasses, and glass-ceramics, and reviews the new microscopes for characterizing materials, and gives an overview of inorganic materials such as zeolites. The theory for XRD texture analysis and analytical methods are also covered. The book is not only up to date on these techniques but also on applications to inorganic materials, both amorphous and crystalline, such as glasses, glass-ceramics, and ceramics.

This book presents scanning electron microscopy (SEM) fundamentals and applications for nanotechnology. It includes integrated fabrication techniques using the SEM, such as e-beam and FIB, and it covers in-situ nanomanipulation of materials. The book is written by international experts from the top nano-research groups that specialize in nanomaterials characterization. The book will appeal to nanomaterials researchers, and to SEM development specialists.

Electron microscopy is frequently portrayed as a discipline that stands alone, separated from molecular biology, light microscopy, physiology, and biochemistry, among other disciplines. It is also presented as a technically demanding discipline operating largely in the sphere of "black boxes" and governed by many absolute laws of procedure. At the introductory level, this portrayal does the discipline and the student a disservice. The instrumentation we use is complex, but ultimately understandable and, more importantly, repairable. The procedures we employ for preparing tissues and cells are not totally understood, but enough information is available to allow investigators to make reasonable choices concerning the best techniques to apply to their parti cular problems. There are countless specialized techniques in the field of electron and light microscopy that require the acquisition of specialized knowledge, particularly for interpretation of results (electron tomography and energy dispersive spectroscopy immediately come to mind), but most laboratories possessing the equipment to effect these approaches have specialists to help the casual user. The advent of computer operated electron microscopes has also broadened access to these instruments, allowing users with little technical knowledge about electron microscope design to quickly become operators. This has been a welcome advance, because earlier instru ments required a level of knowledge about electron optics and vacuum systems to produce optimal photographs and to avoid "crashing" the instruments that typically made it difficult for beginners."

Deals with both the ultrashort laser-pulse technology in the few- to mono-cycle region and the laser-surface-controlled scanning-tunneling microscopy (STM) extending into the spatiotemporal extreme technology. The former covers the theory of nonlinear pulse propagation beyond the slowly-varing-envelope approximation, the generation and active chirp compensation of ultrabroadband optical pulses, the amplitude and phase characterization of few- to mono-cycle pulses, and the feedback field control for the mono-cycle-like pulse generation. In addition, the wavelength-multiplex shaping of ultrabroadband pulses, and the carrier-phase measurement and control of few-cycle pulses are described. The latter covers the CW-laser-excitation STM, the femtosecond-time-resolved STM and atomic-level surface phenomena controlled by femtosecond pulses.

The Nobel Prize of 1986 on Sc- ningTunnelingMicroscopysignaled a new era in imaging. The sc- ning probes emerged as a new - strument for imaging with a p- cision suf?cient to delineate single atoms. At ?rst there were two - the Scanning Tunneling Microscope, or STM, and the Atomic Force Mic- scope, or AFM. The STM relies on electrons tunneling between tip and sample whereas the AFM depends on the force acting on the tip when it was placed near the sample. These were quickly followed by the M- netic Force Microscope, MFM, and the Electrostatic Force Microscope, EFM. The MFM will image a single magnetic bit with features as small as 10nm. With the EFM one can monitor the charge of a single electron. Prof. Paul Hansma at Santa Barbara opened the door even wider when he was able to image biological objects in aqueous environments. At this point the sluice gates were opened and a multitude of different instruments appeared. There are signi?cant differences between the Scanning Probe Microscopes or SPM, and others such as the Scanning Electron Microscope or SEM. The probe microscopes do not require preparation of the sample and they operate in ambient atmosphere, whereas, the SEM must operate in a vacuum environment and the sample must be cross-sectioned to expose the proper surface. However, the SEM can record 3D image and movies, features that are not available with the scanning probes.

Much of this book was written during a sabbatical visit by J. C. H. S. to the Max Planck Institute in Stuttgart during 1991. We are therefore grateful to Professors M. Ruhle and A. Seeger for acting as hosts during this time, and to the Alexander von Humbolt Foundation for the Senior Scientist Award which made this visit possible. The Ph. D. work of one of us (J. M. Z. ) has also provided much of the background for the book, together with our recent papers with various collaborators. Of these, perhaps the most important stimulus to our work on convergent-beam electron diffraction resulted from a visit to the National Science Foundation's Electron Microscopy Facility at Arizona State University by Professor R. H(lJier in 1988, and from a return visit to Trondheim by J. C. H. S. in 1990. We are therefore particularly grateful to Professor H(lJier and his students and co-workers for their encouragement and collaboration. At ASU, we owe a particular debt of gratitude to Professor M. O'Keeffe for his encouragement. The depth of his under standing of crystal structures and his role as passionate skeptic have frequently been invaluable. Professor John Cowley has also been an invaluable sounding board for ideas, and was responsible for much of the experimental and theoretical work on coherent nanodiffraction. The sections on this topic derive mainly from collaborations by J. C. H. S. with him in the seventies."