Biological Low-Voltage Scanning Electron Microscopy

Field-emission, low-voltage scanning electron microscopy (LVSEM) is a field that has grown tremendously in recent years because is offers the optimal method for viewing complex surfaces at high resolution and in three dimensions. However, even though the instrumentation required to get good results at low beam voltage has become increasingly available, there has been a lag in its application to biological specimens. What seemed to be missing was volume that combined both the theory and practice of using this equipment in an optimal manner with a thorough treatment of biological specimen preparation.

Biological Low-Voltage Scanning Electron Microscopy is the first book to address both of these aspects of biological LVSEM. After providing a thorough description of the unique advantages and the operating constraints related to operating a scanning electron microscope at low beam voltage, the remainder of book focuses on the the best way to image all types of plant and animal cells and covers specimens that range from macromolecules to the surfaces revealed by de-embedding resin-embedded samples. Advanced specimen preparation techniques such as cryo-LVSEM, and immuno-gold-LVSEM are fully covered, as is x-ray microanalysis at low beam voltage and live-time stereo imaging. The preparative protocols provided represent the distilled essence of the experience of a group of world-renowned authors who have, for many decades, been instrumental in developing and applying new approaches to LVSEM to support their own biological research.

Résumé

Major improvements in instrumentation and specimen preparation have brought SEM to the fore as a biological imaging technique. In FESEM, a field-emission cathode placed in the electron gun of a scanning electron microscope provides narrower probing beams and high electron energy. The result is improved spatial resolution and minimized sample charging and damage. Images produced are less destroyed and have a spatial resolution down to 1.5 nm, three to six times better than conventional SEM.

Although this imaging technique has undergone tremendous developments, it is still poorly represented in the literature, limited to journal articles and chapter s in books. This comprehensive volume is dedicated to the theory and practical applications of FESEM in biological samples. It provides a comprehensive explanation of instrumentation, applications, and protocols, and is intended to teach the reader how to operate such microscopes to obtain the best quality images.

Contenu

The Early Development of the Scanning Electron Microscope.- LVSEM for Biology.- The Aberration-Corrected SEM.- Noise and Its Effects on the Low-Voltage SEM.- High-Resolution, Low Voltage, Field-Emission Scanning Electron Microscopy (HRLVFESEM) Applications for Cell Biology and Specimen Preparation Protocols.- Molecular Labeling for Correlative Microscopy: LM, LVSEM, TEM, EF-TEM and HVEM.- Low kV and Video-Rate, Beam-Tilt Stereo for Viewing Live-Time Experiments in the SEM.- Cryo-SEM of Chemically Fixed Animal Cells.- High-Resolution and Low-Voltage SEM of Plant Cells.- High-Resolution Cryoscanning Electron Microscopy of Biological Samples.- Developments in Instrumentation for Microanalysis in Low-Voltage Scanning Electron Microscopy.