Microprobe Analysis Of Biological Systems

Microprobe Analysis of Biological Systems covers the proceedings of the 1980 Microprobe Analysis of Biological Systems conference held at Battelle Conference Center in Seattle, Washington. Most of the major laboratories in the field of biological microanalysis in the United States, England, Scotland, France, and Germany are represented. The conference presents the findings, theories, techniques, and procedures of the laboratory represented, no matter how tentative and exploratory. This book is divided into four parts encompassing 22 chapters that focus on biological applications of microprobe analysis. The introductory part describes the application of electron microprobe and X-ray microanalyses in studies of epithelial transport, avian salt gland, electrolyte transport, and acrosome reaction. The subsequent part covers the application of microprobe techniques in the analysis of cardiac, skeletal, vascular smooth, and freeze-dried muscles. It also describes a method for obtaining erythrocyte preparations for validating biological microprobe methods and the continuum-fluorescence effect on thick biological tissue. The method using freeze-substitution to localize calcium in quick-frozen tissue for X-ray microanalysis is also explained. The third part of the book tackles the principles, basic features, and applications of electron energy-loss spectroscopy. Discussions on the use of inner-shell signals for a quantitative local microanalysis technique; theoretical study of the energy resolution; and collection efficiency of a magnetic spectrometer are also included. The final part covers the elemental distribution in single erythrocytes using X-ray microanalysis. It also discusses the fundamentals of cryosectioning process for X-ray microanalysis of diffusible elements and the freezing behavior of a number of chemically different gels chosen for their partial resemblance to biological structures. Considerable chapters contain materials and methods, results, discussions, conclusions, and references. This book will be of value to scientists interested in elemental and ion transport within cells and between cells and extracellular compartments.

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.

The modem microbiologist is often a real specialist who has difficulty under standing and applying many of the techniques beyond those in his or her own immediate field. On the other hand, most benefits to modem microbiology are obtained when a broad spectrum of scientific approaches can be focused on a problem. In early studies, electron microscopy was pivotal in understanding bacterial and viral morphology, and we still feel that we will understand a disease better if we have seen an electron micrograph of the causative agent. Today, because there is an increased awareness of the need to understand the rela tionships between microbial structure and function, the electron microscope is still one of the most important tools microbiologists can use for detailed analysis of microorganisms. Often, however, the aforementioned modem microbiologist still thinks of ultrastructure as involving negative staining or ultrathin sectioning in order to get a look at the shape of a "bug. " Many of the newer ultrastructure techniques, such as gold-labeled antibody localization, freeze-fracture, X-ray microanalysis, enzyme localization, and even scanning electron microscopy, are poorly under stood by, and therefore forbidding to, the average microbiologist. Even many cell biologists admit to having difficulty staying in touch with current develop ments in the fast-moving field of electron microscopy techniques.

Biomedical Applications of Microprobe Analysis is a combination reference/laboratory manual for the use of microprobe analysis in both clinical diagnostic and research settings. Also called microchemical microscopy, microprobe analysis uses high-energy bombardment of cells and tissue, in combination with high resolution EM or confocal microscopy to provide a profile of the ion, metal, and mineral concentrations present in a sample. This allows insight into the physiology and pathophysiology of a wide variety of cells and tissues. This book describes methods for obtaining detailed information about the identity and composition of particles too small to be seen with the naked eye and describes how this information can be useful in diagnostic and biomedical research. Up-to-date review of electron microprobe analysis Detailed descriptions of sample preparation techniques Recent technologies including confocal microscopy, infrared microspectroscopy, and laser raman spectroscopy Over 100 illustrations with numerous specific applications Contributions by world-renowned experts in the field Brief summary of highlights precedes each chapter

Microbeam Analysis in Biology contains the proceedings of a workshop on Biological X-Ray Microanalysis by Electron Beam Excitation, held in Boston, Massachusetts on August 25-26, 1977. This book focuses on the principles, techniques, and biological use of electron probe microanalysis, energy-loss spectroscopy, and ion probe microanalysis. This text reflects the emphasis of the workshop on presenting the principles of analysis, the optimization of operating conditions, the description of successful techniques for sample preparation and quantitation, the illustration of problems and pitfalls, and the direction of microbeam analysis in biology.

This volume is a continuation of two prior books on advanced electron microscope techniques. The purpose of this series has been to provide in depth analyses of methods which are considered to be at the leading edge of electron microscopic research procedures with applications in the biological sciences. The mission of the present volume remains that of a source book for the research practitioner or advanced student, especially one already well versed in basic electron optical methods. It is not meant to provide in troductory material, nor can this modest volume hope to cover the entire spectrum of advanced technology now available in electron microscopy. In the past decade, computers have found their way into many research laboratories thanks to the enormous increase in computing power and stor age available at a modest cost. The ultrastructural area has also benefited from this expansion in a number of ways which will be illustrated in this volume. Half of the contributions discuss technologies that either directly or indirectly make extensive use of computer methods.

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.