This book covers the fundamental principles of fluorescence and their application to fluorescence microscopy, and presents applications to immunofluorescence, in situ hybridization, and photomicrography. It provides troubleshooting guidance to guide the user through commonly encountered problems.

Electron Microscopy and Analysis 1997 celebrates the centenary anniversary of the discovery of the electron by J.J. Thomson in Cambridge and the fiftieth anniversary of this distinguished Institute group. The book includes papers on the early history of electron microscopy (from P. Hawkes), the development of the scanning electron microscope at Cambridge (from K. Smith), electron energy loss spectroscopy (from L.M. Brown), imaging methods (from J. Spence), and the future of electron microscopy (from C. Humphreys). Covering a wide range of applications of advanced techniques, it discusses electron imaging, electron energy-loss and x-ray analysis, and scanning probe and electron beam microscopies. This volume is a handy reference for professionals using microscopes in all areas of physics, materials science, metallurgy, and surface science to gain an overview of developments in our understanding of materials microstructure and of advances in microscope interrogation techniques.

This text describes the interactions of light and the specimen in the microscope in relation to the manner in which they are used to generate contrast in the image of a specimen. Chapters cover bright-field and dark-ground techniques for both transmitted and reflected light, as well as the effects of the refractive index of the mounting medium surrounding the specimen. Subsequent chapters deal with the use of polarized light, phase and modulation contrast, interference and fluorescence and contrast technique outside the microscope.

This work provides detailed practical methods for specimen preparation for negative staining and cryoelectron microscopy of thinly spread samples of biological particulates. The different possibilities are emphasized and the benefits to be gained by the various techniques clearly explained. A wide range of biological applications are presented.

Spectroscopy and spectrofluorometry are core techniques used throughout biology and medicine. These techniques evolve continuously and this book provides information on the latest advances in spectroscopic methods. Light Spectroscopy begins by describing the basic principles and then provides practical guidance on the wide range of current techniques, their application, and analysis of the results obtained.

The First Book on CRS Microscopy Compared to conventional Raman microscopy, coherent Raman scattering (CRS) allows label-free imaging of living cells and tissues at video rate by enhancing the weak Raman signal through nonlinear excitation. Edited by pioneers in the field and with contributions from a distinguished team of experts, Coherent Raman Scattering Microscopy explains how CRS can be used to obtain a point-by-point chemical map of live cells and tissues. In color throughout, the book starts by establishing the foundation of CRS microscopy. It discusses the principles of nonlinear optical spectroscopy, particularly coherent Raman spectroscopy, and presents the theories of contrast mechanisms pertinent to CRS microscopy. The text then provides important technical aspects of CRS microscopy, including microscope construction, detection schemes, and data analyses. It concludes with a survey of applications that demonstrate how CRS microscopy has become a valuable tool in biomedicine. Due to its label-free, noninvasive examinations of living cells and organisms, CRS microscopy has opened up exciting prospects in biology and medicine-from the mapping of 3D distributions of small drug molecules to identifying tumors in tissues. An in-depth exploration of the theories, technology, and applications, this book shows how CRS microscopy has impacted human health and will deepen our understanding of life processes in the future.

Recent advances in microscopy, together with developments in fibre technology and changing commercial priorities, point to the need for an up-to-date manual which details the techniques available. This book is a practical guide to the properties and characteristics of textile fibres, with advice on sampling, specimen preparation and examination procedures.

The contents of the letters published here, again show the great range of subjects that occupied Van Leeuwenhoek: from sugar candy, the shape and crystal structure of diamonds, the dissolution of silver crystals in aqua fortis to gold dust from Guinea dissolved in aqua regia and the dissolution and separation of gold, silver, and copper.

Every volume in the Series contains the texts in the original Dutch and an English translation. The great range of subjects studied by Van Leeuwenhoek is reflected in these letters: instruments to measure water, pulmonary diseases; experiments relating to the solution of gold and silver; salt crystals and grains of sand; botanical work, such as duckweed and germination of orange pips; description on protozoa. blood, spermatozoa and health and hygiene, for example and harmfulness of tea and coffee and the benefits of cleaning teeth.

Although many significant advances have been made in biological specimen preparation during the past 20 years no new practical guide to the techniques has been published in this time. As a result of the recent resurgence of interest in light microscopy, particularly confocal techniques, this up-to-date book should benefit both the professional and amateur alike.

As the selection of material for particular engineering properties becomes increasingly important in keeping costs down, methods for evaluating material properties also become more relevant. One such method examines the geometry of grain boundaries, which reveals much about the properties of the material.
Studying material properties from their geometrical measurements, The Measurement of Grain Boundary Geometry provides a framework for a specialized application of electron microscopy for metals and alloys and, by extension, for ceramics, minerals, and semiconductors. The book presents an overview of the developments in the theory of grain boundary geometry and its practical applications in material engineering. It also covers the tunneling electron microscope (TEM), experimental aspects of data collection, data processing, and examples from actual investigations. Each step of the analysis process is clearly described, from data collection through processing, analysis, representation, and display to applications. The book also includes a glossary of terms.
Exploring both the experimental and analytical aspects of the subject, this practical reference guide is essential for researchers and students involved in material properties, whether in physics, materials science, metallurgy, or physical chemistry.

Quantitative Microbeam Analysis provides a comprehensive introduction to the field of quantitative microbeam analysis (MQA). MQA is a technique used to analyze subatomic quantities of materials blasted from a surface by a laser or particle beam, providing information on the structure and composition of the material. Contributed to by international experts, the book is unique in the breadth of microbeam analytical techniques covered. For each technique, it develops the theoretical background, discusses practical details relating to choice of equipment, and describes the current advances. The book highlights developments relating to Auger electron spectroscopy in scanning electron microscopes and transmission electron microscopes and advances in surface analytical imaging and accelerated ion beam-surface interactions.

Despite substantial evidence showing the feasibility of Atomic Force Microscopy (AFM) to identify cells with altered elastic and adhesive properties, the use of this technique as a complementary diagnostic method remains controversial. This book is designed to be a practical textbook that teaches how to assess the mechanical characteristics of living, individual cells by AFM. Following a step-by-step approach, it introduces the methodology of measurements in the case of both determination of elastic properties and quantification of adhesive properties.

This text details the practical steps which must precede microscopy. Methods for preparing sheet or disk specimens and final thinning techniques are described with reference to practical problems. It also covers methods for mounting speciments in the TEM, and guides the reader on the most appropriate choice of speciment-preparation method.

How deep we can see inside Nature's smallest secrets? Will it be possible some day in the near future to investigate living structures at atomic level? This area of study is very interdisciplinary, since it applies the principles and the techniques of biology, physics, chemistry, mathematics, and engineering to elucidate the structures of biological macromolecules, of supramolecular structures, organelles, and cells. This book offers updated information on how much information we are able to obtain in the exploration of the inner details of biological specimens in their native structure and composition. The book deals with the implementation of laser beam and stage scanning systems incorporating confocal optics or multiphoton microscopy; the advent of new electro-optical detectors with great sensitivity, linearity, and dynamic range; the possibility of 2D fast image enhancement, reconstruction, restoration, analysis and 3D display, and the application of luminescence techniques (FLIMT, FRET combined with the use of quantum dots), which gives the possibility to investigate the chemical and molecular spatio-temporal organization of life processes; Electron Microscopy and Scanning Force Microscopy (SFM), are also presented, which has opened completely new perspectives for analyzing the surface topography of biological matter in its aqueous environment at a resolution comparable to that achieved by EM.

Uses questions about hypothetical situations to introduce the process of thinking according to scientific method.

Optical Imaging Techniques in Cell Biology, Second Edition covers the field of biological microscopy, from the optics of the microscope to the latest advances in imaging below the traditional resolution limit. It includes the techniques-such as labeling by immunofluorescence and fluorescent proteins-which have revolutionized cell biology. Quantitative techniques such as lifetime imaging, ratiometric measurement, and photoconversion are all covered in detail. Expanded with a new chapter and 40 new figures, the second edition has been updated to cover the latest developments in optical imaging techniques. Explanations throughout are accurate, detailed, but as far as possible non-mathematical. This edition includes appendices with useful practical protocols, references, and suggestions for further reading. Color figures are integrated throughout.

Computer Techniques for Image Processing in Electron Microscopy, Volume 214 in the Advances in Imaging and Electron Physics series, presents the latest advances in the field, with this new volume covering Image Formation Theory, The Discrete Fourier Transform, Analytic Images, The Image and Diffraction Plane Problem: Uniqueness, The Image and Diffraction Plane Problem: Numerical Methods, The Image and Diffraction Plane Problem: Computational Trials, Alternative Data for the Phase Determination, The Hardware of Digital Image Handling, Basic Software or Digital Image Handling, Improc, and much more.

Stereology, or quantitative microscopy, is a basic research tool in science and technology. The emergence of design-based methods has greatly increased the power, flexibility, adaptability, and scope of stereology applications, establishing a closer connection between statistics and quantitative microscopy. Despite its scientific importance, modern stereology remains largely unknown to the statistical community, with valuable information either widely scattered or inaccessible to newcomers to the field. Now is the perfect time for a book that enables biostatisticians and statistical consultants to give beneficial advice to researchers in microscopy.

Stereology for Statisticians sets out the principles of stereology from a statistical viewpoint, focusing on both basic theory and practical implications. This book discusses ways to effectively communicate statistical issues to clients, draws attention to common methodological errors, and provides references to essential literature.

The first full text on design-based stereology, it opens with a review of classical and modern stereology, followed by a treatment of mathematical foundations such as geometry, probability, and statistical inference. The book then presents core techniques, including estimation of absolute geometrical quantities, relative quantities, and statistical inference for populations of discrete objects. The final chapters discuss implementing techniques in practical sampling designs, summarize understanding of the variance of stereological estimators, and describe open problems for further research.

Making a clear distinction is made between nano- and micro-mechanical testing for physical reasons, this monograph describes the basics and applications of the supermicroscopies AFM and SNOM, and of the nanomechanical testing on rough and technical natural surfaces in the submicron range down to a lateral resolution of a few nm. New or improved instrumentation, new physical laws and unforeseen new applications in all branches of natural sciences (around physics, chemistry, mineralogy, materials science, biology and medicine) and nanotechnology are covered as well as the sources for pitfalls and errors. It outlines the handling of natural and technical samples in relation to those of flat standard samples and emphasizes new special features. Pitfalls and sources of errors are clearly demonstrated as well as their efficient remedy when going from molecularly flat to rough surfaces. The academic or industrial scientist learns how to apply the principles for tackling their scientific or manufacturing tasks that include roughness far away from standard samples.

This book focuses primarily on the atomic force microscope and serves as a reference for students, postdocs, and researchers using atomic force microscopes for the first time. In addition, this book can serve as the primary text for a semester-long introductory course in atomic force microscopy. There are a few algebra-based mathematical relationships included in the book that describe the mechanical properties, behaviors, and intermolecular forces associated with probes used in atomic force microscopy. Relevant figures, tables, and illustrations also appear in each chapter in an effort to provide additional information and points of interest. This book includes suggested laboratory investigations that provide opportunities to explore the versatility of the atomic force microscope. These laboratory exercises include opportunities for experimenters to explore force curves, surface roughness, friction loops, conductivity imaging, and phase imaging.

Electron Microscopy of Interfaces in Metals and Alloys examines the structure of interfaces in metals and alloys using transmission electron microscopy. The book presents quantitative methods of analysis and reviews the most significant work on interface structure over the last 20 years. It provides the first book description of the methods used for quantitative identification of Burgers vectors of interfacial dislocations, including the geometric analysis of periodicities in interface structure and the comparison of experimental and theoretical electron micrographs. The book explores low- and high-angle grain boundaries and interphase interfaces between neighboring grains, emphasizing interfacial dislocations and rigid-body displacements to the structure and properties of interfaces. It also analyzes the use of two-beam images and diffraction patterns for analysis and studies n-beam lattice imaging. The book includes numerous worked examples of the analysis of the structure of grain boundaries and interphaseinterfaces, which are particularly useful to those who need to consider the nature of intercrystalline interfaces.

This work is based on experiences acquired by the authors regarding often asked questions and problems during manifold education of beginners in analytical transmission electron microscopy. These experiences are summarised illustratively in this textbook. Explanations based on simple models and hints for the practical work are the focal points. This practically- oriented textbook represents a clear and comprehensible introduction for all persons who want to use a transmission electron microscope in practice but who are not specially qualified electron microscopists up to now.

Developments in cryo-electron microscopy are creating new opportunities within structural biology and there is currently great interest in developing cryo-EM as a core tool for atomic level structural biology. Many structural techniques can give atomic or near atomic level information, but lack the ability to study proteins within a near-native environment, for example within a cellular compartment. Cryo-EM provides this opportunity, but despite the recent massive improvements in single particle cryo-EM, obtaining sub-2A structural information is still a major challenge. Cryo-electron microscopy has undergone significant developments in microscope design, camera technology and data processing regimes, but there are significant challenges that remain and opportunities to explore, many of which must be tackled by the community as a whole, rather than by individual groups. For example, sample preparation is central to electron microscopy and is currently a significant bottleneck in many experiments, and there are significant problems with ensuring the integrity of the field in terms of dealing with inherently low signal-to-noise images. This volume brings together leading researchers from the UK and the international cryo-electron microscopy community to discuss current developments and new challenges in the field. In this volume the topics covered include: Sample preparation in single particle cryo-EM Pushing the limits in single particle cryo-EM Tomographic analysis, CLEM Map/model validation and machine learning in EM

Super-Resolution Microscopy Techniques in the Neurosciences serves as a comprehensive description of current super-resolution techniques, including the physical principles that allowed for their development, some of the most recent neurobiological applications and selected information for the practical use of these technologies. Written for the Neuromethods series, this detailed work contains contributions from experts in the field and provides key implementation advice to ensure successful results in the lab. Authoritative and cutting-edge, Super-Resolution Microscopy Techniques in the Neurosciences is an ideal guide for researchers aiming to continue increasing the resolution in the imaging capabilities of neuroscientists and thereby changing the perspective in which cellular biology processes are understood at the nanometer scale.

This third edition of Electron Microscopy: Methods and Protocols expands upon the previous editions with current, detailed protocols on biological and molecular research techniques based on TEM and SEM as well as other closely related imaging and analytical methods. With new chapters on conventional and microwave assisted specimen, cryo-specimen preparation, negative staining and immunogold labelling techniques, DNA and RNA tracking using hybrization in TEM or Atomic Force Microscopy, TEM crystallography and cryo TEM 3D tomography, 3D tomography of resin embedded tissues using FIB-SEM, Correlative microscopy using fluorescence microscopy, confocal microscopy or immune labelling techniques for both TEM and FIB-SEM and Elemental and isotopic identification and their distribution in cells and tissues using TEM, SEM, Scanning Transmission Electron Microscopy (STEM), Secondary Ion Mass Spectrometry (SIMS) and Nano SIMS. Written in the highly 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 laboratory protocols and tips on troubleshooting and avoiding known pitfalls. Authoritative and practical, Electron Microscopy: Methods and Protocols, Third Edition provides the most up-to-date and essential information in electron microscopy techniques and methods provided in this edition will assist in advancing future molecular and biological research.