Investigations of frozen-hydrated biological samples using the transmission electron microscope is a challenging task. In cell and structural biology large data sets of consistent quality are required. To record the necessary amount of micrographs in an efficient and timely manner, automated procedures are essential to control the electron microscope. The aim of this work was the development of a software package that enables the user to acquire micrographs of frozen-hydrated biological samples in an automated and unsupervised way for single-particle and tomography applications. The foundations of automation of an electron microscope and the necessary algorithms are presented, furthermore the application of the software is illustrated based on examples from single particle investigations and tomography.
Three-Dimensional Electron Microscopy, Volume 152 in the Methods in Cell Biology series, highlights new advances in the field, with this new volume presenting interesting chapters focusing on FIB-SEM of mouse nervous tissue: fast and slow sample preparation, Serial-section electron microscopy using ATUM - Automated Tape collecting Ultra-Microtome, Software for automated acquisition of electron tomography tilt series, Scanning electron tomography of biological samples embedded in plastic, Cryo-STEM tomography for Biology, CryoCARE: Content-aware denoising of cryo-EM images and tomograms using artificial neural networks, Expedited large-volume 3-D SEM workflows for comparative vertebrate microanatomical imaging, and many other interesting topics. Provides the authority and expertise of leading contributors from an international board of authors Presents the latest release in the Methods in Cell Biology series Includes the latest information on the Three-Dimensional Electron Microscopy technique
Volume Electron Microscopy (vEM), Volume 177 is a collective term for a set of three-dimensional high-resolution ultrastructural imaging techniques that have delivered new insights into biological systems in recent years, garnering substantial interest in the life and clinical sciences. In this book, users will find a variety of vEM workflows and technologies, highlighting application areas with biologically relevant examples. Topics covered include Automated large volume sample preparation for vEM, Resin comparison for Serial Block Face Scanning Volume Electron Microscopy, Immunolabelling for SBF-SEM, Electron Microscopy in Plants, Serial section electron tomography, Automated Tape Collecting Ultramicrotomy (ATUM) for targeting neuropathology, Array tomography, and much more. Other sections focus on Mitochondria morphometry in 3d datasets of mouse brain obtained with serial block-face Scanning Electron Microscopy, Serial Block-face Scanning Electron Microscopy of Schmidtea mediterranea, Correlative multiscale microCT-SBF-SEM imaging of resin-embedded tissue, Methods of enhanced FIB-SEM sample preparation and image acquisition, Functional characterization of endo-lysosomal compartments by correlative live-cell and volume electron microscopy, and much more. Includes chapters written by key leaders and developers in the field Provides detailed protocols, allowing for the application of workflows in one’s own laboratory setting Presents real tips and tricks you won't get from standard research papers
Cryoelectron microscopy of biological molecules is among the hottest growth areas in biophysics and structural biology at present, and Frank is arguably the most distinguished practitioner of this art. CryoEM is likely over the next few years to take over much of the structural approaches currently requiring X-ray crystallography, because one can now get good and finely detailed images of single molecules down to as little as 200,000 MW, covering a substantial share of the molecules of greatest biomedical research interest. This book, the successor to an earlier work published in 1996 with Academic Press, is a natural companion work to our forthcoming book on electron crystallography by Robert Glaeser, with contributions by six others, including Frank. A growing number of workers will employ CryoEM for structural studies in their own research, and a large proportion of biomedical researchers will have a growing interest in understanding what the capabilities and limits of this approach are.
Three-Dimensional Electron Microscopy of Macromolecular Assemblies is the first systematic introduction to single-particle methods of reconstruction. It covers correlation alignment, classification, 3D reconstruction, restoration, and interpretation of the resulting 3D images in macromolecular assemblies. It will be an indispensable resource for newcomers to the field and for all using or adopting these methods. Key Features* Presents methods that offer an alternative to crystallographic techniques for molecules that cannot be crystallized* Describes methods that have been instrumental in exploring the three-dimensional structure of* the nuclear pore complex* the calcium release channel;* the ribosome* chaperonins
This volume discusses different approaches to workflows for large volume electron microscopy - from preparation of samples to their imaging in a variety of microscopes - in some cases also applying correlative techniques. The chapters in this book cover topics such as correlative super resolution and electron microscopy to detect molecules in their native cellular context; low-threshold access to serial section arrays; improving serial blockface SEM by focal charge compensation; FIBSEM analysis of interfaces between hard technical devices and soft neuronal tissue; and image processing for volume electron microscopy. In Neuromethods series style, chapters include the kind of detail and key advice from the specialists needed to get successful results in your laboratory. Cutting-edge and authoritative, Volume Microscopy: Multiscale Imaging with Photons, Electrons, and Ions is a valuable resource for novice and expert scientists interested in learning more about this evolving field.
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.
Brings a fresh point of view to the current state of correlative imaging and the future of the field This book provides contributions from international experts on correlative imaging, describing their vision of future developments in the field based on where it is today. Starting with a brief historical overview of how the field evolved, it presents the latest developments in microscopy that facilitate the correlative workflow. It also discusses the need for an ideal correlative probe, applications in proteomic and elemental analysis, interpretation methods, and how correlative imaging can incorporate force microscopy, soft x-ray tomography, and volume electron microscopy techniques. Work on placing individual molecules within cells is also featured. Correlative Imaging: Focusing on the Future offers in-depth chapters on: correlative imaging from an LM perspective; the importance of sample processing for correlative imaging; correlative light and volume EM; correlation with scanning probe microscopies; and integrated microscopy. It looks at: cryo-correlative microscopy; correlative cryo soft X-ray imaging; and array tomography. Hydrated-state correlative imaging in vacuo, correlating data from different imaging modalities, and big data in correlative imaging are also considered. Brings a fresh view to one of the hottest topics within the imaging community: the correlative imaging field Discusses current research and offers expert thoughts on the field’s future developments Presented by internationally-recognized editors and contributors with extensive experience in research and applications Of interest to scientists working in the fields of imaging, structural biology, cell biology, developmental biology, neurobiology, cancer biology, infection and immunity, biomaterials and biomedicine Part of the Wiley–Royal Microscopical Society series Correlative Imaging: Focusing on the Future will appeal to those working in the expanding field of the biosciences, correlative microscopy and related microscopic areas. It will also benefit graduate students working in microscopy, as well as anyone working in the microscopy imaging field in biomedical research.
Biophysics is a rapidly-evolving interdisciplinary science that applies theories and methods of the physical sciences to questions of biology. Biophysics encompasses many disciplines, including physics, chemistry, mathematics, biology, biochemistry, medicine, pharmacology, physiology, and neuroscience, and it is essential that scientists working in these varied fields are able to understand each other's research. Comprehensive Biophysics, Nine Volume Set will help bridge that communication gap. Written by a team of researchers at the forefront of their respective fields, under the guidance of Chief Editor Edward Egelman, Comprehensive Biophysics, Nine Volume Set provides definitive introductions to a broad array of topics, uniting different areas of biophysics research - from the physical techniques for studying macromolecular structure to protein folding, muscle and molecular motors, cell biophysics, bioenergetics and more. The result is this comprehensive scientific resource - a valuable tool both for helping researchers come to grips quickly with material from related biophysics fields outside their areas of expertise, and for reinforcing their existing knowledge. Biophysical research today encompasses many areas of biology. These studies do not necessarily share a unique identifying factor. This work unites the different areas of research and allows users, regardless of their background, to navigate through the most essential concepts with ease, saving them time and vastly improving their understanding The field of biophysics counts several journals that are directly and indirectly concerned with the field. There is no reference work that encompasses the entire field and unites the different areas of research through deep foundational reviews. Comprehensive Biophysics fills this vacuum, being a definitive work on biophysics. It will help users apply context to the diverse journal literature offering, and aid them in identifying areas for further research Chief Editor Edward Egelman (E-I-C, Biophysical Journal) has assembled an impressive, world-class team of Volume Editors and Contributing Authors. Each chapter has been painstakingly reviewed and checked for consistent high quality. The result is an authoritative overview which ties the literature together and provides the user with a reliable background information and citation resource
