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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.
This book presents a wide variety of immuno-gold techniques for use
in virus diagnosis and research. Protocols are presented for
state-of-the-art techniques, including in situ hybridization,
freeze substitution, and the utilization of ultra-small probes and
replicas for use by virologists and electron microscopists
identifying and studying viruses, their components, and replication
in cells. The procedures are described by eminent scientists and
are pertinent to both experienced researchers and newcomers to this
field who are interested in the localization of low antigenic mass
structures.
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