The development of field-emission scanning electron microscopy for imaging biological surfaces.
This article traces the important milestones in the development of high-resolution, field-emission, scanning electron microscopes (SEM). Such instruments are now capable of producing images of the surfaces of biological specimens that rival, in terms of resolution and contrast, those produced by conventional transmission electron microscopy (TEM). Even though one of the first instruments to produce a useful transmission electron microscope image was, in fact, an early scanning microscope, TEM reached its full potential for biological imaging almost 30 years sooner than did SEM. The main reason for this slow rate of development is the dependence of any scanning technique on source brightness. The only suitable electron source was the field-emission source, originally developed in the 1930's. Making this into a stable and reliable electron source for microscopy required many technical barriers to be overcome. An additional delay may have been caused by the great success that attended the introduction of early SEM instruments. These instruments which employed heated, tungsten hairpin cathodes, were inexpensive and reliable, but they that were also far from optimal in terms of optical performance. Their market success may have engendered the sense of inertia and complacency that further delayed the introduction of low aberrations objective lenses and field-emission sources for almost 20 years after they were first introduced to electron microscopy. In addition, the fact that these early SEMs accustomed users to operating with a much higher beam voltage than was either necessary or wise, lead many to assume that the SEM was incapable of producing high-resolution images of biological surfaces. This left them open to fascination with newer ahd slower techniques that, on balance, were less suitable than optimized SEM for most of their imaging needs. In parallel to these developments in instrumentation, major improvements were also made in the way that the specimen surface was prepared before placing it into the vacuum and radiation environment of the microscope.
Pawley
J
J
Zoology Department, University of Wisconsin-madison 53706, USA.
D002462:Cell Membrane / Q000648:ultrastructure*; D008855:Microscopy, Electron, Scanning / Q000295:instrumentation* / Q000379:methods*; D013499:Surface Properties