Download or read book Cryopreparation of Thin Biological Specimens for Electron Microscopy written by Norbert Roos and published by Oxford University Press, USA. This book was released on 1990 with total page 116 pages. Available in PDF, EPUB and Kindle. Book excerpt: Cryotechniques offer a number of advantages over conventional electron microscopy methods in biology, particularly with regard to preserving the morphology and composition of specimens. This handbook provides a practical guide for researchers, students, and technicians who wish to employ cryotechniques in their work. It covers those methods where the cryofixed specimen is sectioned or otherwise presented as a thin film for examination, giving special attention to topics of current interest. This guide will be of use to workers in a variety of biomedical disciplines, including cell biology, molecular biology, bacteriology, and pathology.

Download or read book Cryotechniques in Biological Electron Microscopy written by Rudolf A. Steinbrecht and published by Springer Science & Business Media. This book was released on 2012-12-06 with total page 308 pages. Available in PDF, EPUB and Kindle. Book excerpt: To preserve tissue by freezing is an ancient concept going back pre sumably to the practice of ice-age hunters. At first glance, it seems as simple as it is attractive: the dynamics of life are frozen in, nothing is added and nothing withdrawn except thermal energy. Thus, the result should be more life-like than after poisoning, tan ning and drying a living cell as we may rudely call the conventional preparation of specimens for electron microscopy. Countless mishaps, however, have taught electron microscopists that cryotechniques too are neither simple nor necessarily more life-like in their outcome. Not too long ago, experts in cryotechniques strictly denied that a cell could truly be vitrified, i.e. that all the solutes and macro molecules could be fixed within non-crystalline, glass-like solid water without the dramatic shifts and segregation effects caused by crystallization. We now know that vitrification is indeed pos sible. Growing insight into the fundamentals of the physics of water and ice, as well as increasing experience of how to cool cells rapidly enough have enlivened the interest in cryofixation and pro duced a wealth of successful applications.

Download or read book Sample Preparation Handbook for Transmission Electron Microscopy written by Jeanne Ayache and published by Springer Science & Business Media. This book was released on 2010-06-08 with total page 338 pages. Available in PDF, EPUB and Kindle. Book excerpt: Successful transmission electron microscopy in all of its manifestations depends on the quality of the specimens examined. Biological specimen preparation protocols have usually been more rigorous and time consuming than those in the physical sciences. For this reason, there has been a wealth of scienti c literature detailing speci c preparation steps and numerous excellent books on the preparation of b- logical thin specimens. This does not mean to imply that physical science specimen preparation is trivial. For the most part, most physical science thin specimen pre- ration protocols can be executed in a matter of a few hours using straightforward steps. Over the years, there has been a steady stream of papers written on various aspects of preparing thin specimens from bulk materials. However, aside from s- eral seminal textbooks and a series of book compilations produced by the Material Research Society in the 1990s, no recent comprehensive books on thin specimen preparation have appeared until this present work, rst in French and now in English. Everyone knows that the data needed to solve a problem quickly are more imp- tant than ever. A modern TEM laboratory with supporting SEMs, light microscopes, analytical spectrometers, computers, and specimen preparation equipment is an investment of several million US dollars. Fifty years ago, electropolishing, chemical polishing, and replication methods were the principal specimen preparation me- ods.

Download or read book Biological Specimen Preparation for Transmission Electron Microscopy written by Audrey M. Glauert and published by Princeton University Press. This book was released on 2014-07-14 with total page 349 pages. Available in PDF, EPUB and Kindle. Book excerpt: This book contains all the necessary information and advice for anyone wishing to obtain electron micrographs showing the most accurate ultrastructural detail in thin sections of any type of biological specimen. The guidelines for the choice of preparative methods are based on an extensive survey of current laboratory practice. For the first time, in a textbook of this kind, the molecular events occurring during fixation and embedding are analysed in detail. The reasons for choosing particular specimen preparation methods are explained and guidance is given on how to modify established techniques to suit individual requirements. All the practical methods advocated are clearly described, with accompanying tables and the results obtainable are illustrated with many electron micrographs. Portland Press Series: Practical Methods in Electron Microscopy, Volume 17, Audrey M. Glauert, Editor Originally published in 1999. The Princeton Legacy Library uses the latest print-on-demand technology to again make available previously out-of-print books from the distinguished backlist of Princeton University Press. These editions preserve the original texts of these important books while presenting them in durable paperback and hardcover editions. The goal of the Princeton Legacy Library is to vastly increase access to the rich scholarly heritage found in the thousands of books published by Princeton University Press since its founding in 1905.

Download or read book na written by and published by . This book was released on with total page pages. Available in PDF, EPUB and Kindle. Book excerpt:

Download or read book Specimen Preparation for Transmission Electron Microscopy of Materials written by PJ Goodhew and published by Garland Science. This book was released on 2020-11-25 with total page 50 pages. Available in PDF, EPUB and Kindle. Book excerpt: Details the essential practical steps which must precede microscopy. Methods for preparing sheet or disc specimens and final thinning techniques are described with reference to practical problems. The book also covers methods for mounting specimens in the

Download or read book Preparation of Biological Specimens for Scanning Electron Microscopy written by Judith A. Murphy and published by . This book was released on 1984 with total page 364 pages. Available in PDF, EPUB and Kindle. Book excerpt:

Download or read book Methods of Preparation for Electron Microscopy written by David G. Robinson and published by Springer Science & Business Media. This book was released on 2012-12-06 with total page 204 pages. Available in PDF, EPUB and Kindle. Book excerpt: In 1939, when the electron optics laboratory of Siemens & Halske Inc. began to manufacture the first electron microscopes, the biological and medical profes sions had an unexpected instrument at their disposal which exceeded the reso lution of the light microscope by more than a hundredfold. The immediate and broad application of this new tool was complicated by the overwhelming prob lems inherent in specimen preparation for the investigation of cellular struc tures. The microtechniques applied in light microscopy were no longer appli cable, since even the thinnest paraffin layers could not be penetrated by electrons. Many competent biological and medical research workers expressed their anxiety that objects in high vacuum would be modified due to complete dehydration and the absorbed electron energy would eventually cause degrada tion to rudimentary carbon backbones. It also seemed questionable as to whether it would be possible to prepare thin sections of approximately 0. 5 11m from heterogeneous biological specimens. Thus one was suddenly in posses sion of a completely unique instrument which, when compared with the light microscope, allowed a 10-100-fold higher resolution, yet a suitable preparation methodology was lacking. This sceptical attitude towards the application of electron microscopy in bi ology and medicine was supported simultaneously by the general opinion of colloid chemists, who postulated that in the submicroscopic region of living structures no stable building blocks existed which could be revealed with this apparatus.

Download or read book Cryo electron Microscopy for the Study of Hybrid Materials and Biological Specimens written by Katherine Anne Spoth and published by . This book was released on 2018 with total page 284 pages. Available in PDF, EPUB and Kindle. Book excerpt: Cryo-electron microscopy has grown overwhelmingly in popularity in recent years. Instrumental developments enable higher- and higher-resolution studies of biological molecules to near-atomic resolution using single particle analysis, while cryo-TEM tomography gives three-dimensional information about cellular structure. This thesis applies cryo-EM in less-conventional studies: in applications to fields outside of biology, and studies of cryo-scanning TEM (STEM) techniques with new detector technology. Vitrification methods for cryo-TEM perfectly preserve native structure in solution. It is used here to study the early formation processes of mesoporous silica nanoparticles, which are templated by self-assembling surfactant molecules in solution. Direct imaging of this hybrid inorganic/organic material with organics intact would be difficult without vitrifying the reaction solution. Through cryo-TEM imaging of different stages of the formation process we identify synthesis parameters affecting the properties of the fully-formed nanoparticles: relative reactant concentrations, stirring rate, and types of silica precursors. We study formation of hierarchical structures of single-pore particles, from their initial formation to alignment in a 1D cylinder, higher-order nanosheet, and helical structures. Structural preservation by cryo-EM is also utilized to quantitatively describe a shape change in hexagonal silica particles using cryo-STEM imaging, including three dimensional structure determined using cryo-STEM tomography. STEM has been the technique of choice for high-resolution imaging of materials, however conventional detectors discard much of the incident electron dose making the technique's use difficult for radiation-sensitive cryo-EM specimens. A new direct detector for STEM, the Electron Microscope Pixel Array Detector (EMPAD) records the full diffraction pattern at each scan position, allowing nearly all electrons incident on a specimen to be used for imaging. In particular, we describe a new technique for bright field imaging, where pixelated detection over the central diffraction disk allows collection of many images at different relative tilts to the optical axis. Because the images are detected separately, we can measure the tilt-induced image shift at each pixel and correct for the shifts, creating a coherent image with improved signal-to-noise ratio and resolution over both BF-STEM and conventional TEM imaging. The improvement is greater for thick specimens, for which effects due to chromatic aberrations are greatly reduced in STEM compared to TEM. At very low doses, tcBF-STEM outperforms TEM, which will lead to improvements in tomography where the total dose must be fractioned over all frames in the tilt series.

Download or read book Low Temperature Microscopy and Analysis written by Patrick Echlin and published by Springer Science & Business Media. This book was released on 2013-11-11 with total page 553 pages. Available in PDF, EPUB and Kindle. Book excerpt: The frozen-hydrated specimen is the principal element that unifies the subject of low temperature microscopy, and frozen-hydrated specimens are what this book is all about. Freezing the sample as quickly as possible and then further preparing the specimen for microscopy or microanalysis, whether still embedded in ice or not: there seem to be as many variations on this theme as there are creative scientists with problems of structure and composition to investigate. Yet all share a body of com mon fact and theory upon which their work must be based. Low-Temperature Micros copy and Analysis provides, for the first time, a comprehensive treatment of all the elements to which one needs access. What is the appeal behind the use of frozen-hydrated specimens for biological electron microscopy, and why is it so important that such a book should now have been written? If one cannot observe dynamic events as they are in progress, rapid specimen freezing at least offers the possibility to trap structures, organelles, macro molecules, or ions and other solutes in a form that is identical to what the native structure was like at the moment of trapping. The pursuit of this ideal becomes all the more necessary in electron microscopy because of the enormous increase in resolution that is available with electron-optical instruments, compared to light optical microscopes.

Download or read book Challenges and Solutions in Sample Preparation for High Resolution Cryo Electron Microscopy written by Felix Weis and published by Frontiers Media SA. This book was released on 2022-12-05 with total page 120 pages. Available in PDF, EPUB and Kindle. Book excerpt:

Download or read book Cryo EM Part A Sample Preparation and Data Collection written by and published by Academic Press. This book was released on 2010-09-30 with total page 466 pages. Available in PDF, EPUB and Kindle. Book excerpt: Cryo-EM Part A: Sample Preparation and Data Collection is dedicated to a description of the instruments, samples, protocols, and analyses that belong to cryo-EM. It emphasizes the relatedness of the ideas, instrumentation, and methods underlying all cryo-EM approaches, which allow practitioners to easily move between them. Within each section, the articles are ordered according to the most common symmetry of the sample to which their methods are applied. Includes time-tested core methods and new innovations applicable to any researcher Methods included are useful to both established researchers and newcomers to the field Relevant background and reference information given for procedures can be used as a guide

Download or read book Cryo Electron Tomography written by Friedrich Förster and published by Springer Nature. This book was released on with total page 391 pages. Available in PDF, EPUB and Kindle. Book excerpt:

Download or read book Electron Probe Microanalysis written by Karl Zierold and published by Springer Science & Business Media. This book was released on 2013-03-08 with total page 331 pages. Available in PDF, EPUB and Kindle. Book excerpt: The aim of electron probe microanalysis of biological systems is to identify, localize, and quantify elements, mass, and water in cells and tissues. The method is based on the idea that all electrons and photons emerging from an electron beam irradiated specimen contain information on its structure and composition. In particular, energy spectroscopy of X-rays and electrons after interaction of the electron beam with the specimen is used for this purpose. However, the application of this method in biology and medicine has to overcome three specific problems: 1. The principle constituent of most cell samples is water. Since liquid water is not compatible with vacuum conditions in the electron microscope, specimens have to be prepared without disturbing the other components, in parti cular diffusible ions (elements). 2. Electron probe microanaly sis provides physical data on either dry specimens or fully hydrated, frozen specimens. This data usually has to be con verted into quantitative data meaningful to the cell biologist or physiologist. 3. Cells and tissues are not static but dynamic systems. Thus, for example, microanalysis of physiolo gical processes requires sampling techniques which are adapted to address specific biological or medical questions. During recent years, remarkable progress has been made to overcome these problems. Cryopreparation, image analysis, and electron energy loss spectroscopy are key areas which have solved some problems and offer promise for future improvements.

Download or read book Negative Staining and Cryoelectron Microscopy written by 0 J.R. Harris, and published by Taylor & Francis. This book was released on 2023-05-09 with total page 204 pages. Available in PDF, EPUB and Kindle. Book excerpt: Since the start of biological studies using the transmission electron microscope scientists have sought to develop procedures for the preparation and investigation of the thinly spread specimens of biological particulates. Negative Staining and Cryoelect

Download or read book Cryogenic Electron Microscopy and Tomography on Radiation Sensitive Crystalline Materials written by Weijiang Zhou and published by . This book was released on 2022 with total page pages. Available in PDF, EPUB and Kindle. Book excerpt: Cryogenic electron microscopy (Cryo-EM) has been one of the most exciting technology advancements in biological field in recent years. The idea of freezing samples in liquid nitrogen temperature provides a unique advantage to preserve and observe the object of interest in their native state. The improvements in both electron microscopy related hardware and data processing software enable the retrieval of structural information to atomic details. However, many technical features in electron microscope have not been fully explored for cryo-EM and many other sample systems other than biological specimens could utilize cryo-EM to accelerate their structural characterization research. This dissertation focuses on developing a new cryo-EM based experimental and data processing pipeline to further extend the applications of cryo-EM to broader research fields, where the samples of interest are normally not amenable to conventional electron microscopy methods. There are two common features across the specimens aforementioned: first, they are crystalline materials, meaning they are either single crystals or contain certain degree of periodic structures; second, they are sensitive to electron radiation, thus conventional transmission electron microscopy is not suitable to study such materials due to its high electron radiation damage. To investigate those specimens using cryo-EM, we developed several new methods: customized freezing protocol to preserve the samples from ambient environment; cryogenic focused ion beam milling to prepare electron transparent samples; continuous tilting electron diffraction to solve the atomic structure from single crystals; electron diffraction and high resolution low dose electron imaging to acquire the sub-nanometer structural information in both real-space and Fourier space; electron tomography and automated annotation to recover the morphology of non-biological samples in 3D. Those lead to many unprecedented findings in those representative specimens, which will be described in this dissertation. We anticipate those new technical developments will facilitate the usage of cryo-EM in structural biology, chemical biology, organic chemistry and material science in the near future.

Download or read book Specimen Preparation for Transmission Electron Microscopy of Materials written by Peter J. Goodhew and published by Garland Science. This book was released on 1984 with total page 56 pages. Available in PDF, EPUB and Kindle. Book excerpt: Details the essential practical steps which must precede microscopy. Methods for preparing sheet or disc specimens and final thinning techniques are described with reference to practical problems. The book also covers methods for mounting specimens in the