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The electron microscope is a type of microscope that uses electrons to create an image of the target. It has much higher magnification and resolving power than a normal light microscope, up to two million times, allowing it to see smaller objects and details.
The first electron microscope prototype was built in 1931 by the German engineers Ernst Ruska and Max Knoll [1]. It was based on the ideas and discoveries of French physicist Louis de Broglie. Although it was primitive and not fit for practical use, the instrument was still capable of magnifying objects by four hundred times.
Reinhold Rudenberg, the research director of Siemens, had patented the electron microscope in 1931, although Siemens was doing no research on electron microscopes at that time. In 1937 Siemens began developing the electron microscope, funding Ruska and Bodo von Borries to develop the instrument. Siemens also employed Ruska's brother Helmut to work on applications, particularly with biological materials. [2][3]
Siemens produced the first commercial TEM in 1939, but the first practical electron microscope was built at the University of Toronto in 1938, by Eli Franklin Burton and students Cecil Hall, James Hillier and Albert Prebus.[4]
Although modern electron microscopes can magnify objects up to two million times, they are still based upon Ruska's prototype and his correlation between wavelength and resolution. The electron microscope is an integral part of many laboratories. Researchers use it to examine biological materials (such as microorganisms and cells), a variety of large molecules, medical biopsy samples, metals and crystalline structures, and the characteristics of various surfaces.
Major manufacturers include:
The original form of electron microscopy, Transmission electron microscopy (TEM) involves a high voltage electron beam emitted by a cathode and formed by magnetic lenses. The electron beam that has been partially transmitted through the very thin (and so semitransparent for electrons) specimen carries information about the inner structure of the specimen. The spatial variation in this information (the "image") is then magnified by a series of magnetic lenses until it is recorded by hitting a fluorescent screen, photographic plate, or light sensitive sensor such as a CCD (charge-coupled device) camera. The image detected by the CCD may be displayed in real time on a monitor or computer.
Resolution of the high-resolution TEM (HRTEM) is limited by spherical aberration and chromatic aberration, but a new generation of aberration correctors have been able to overcome spherical aberration. Software correction of spherical aberration has allowed the production of images with sufficient resolution to show carbon atoms in diamond separated by only 0.89 ångström (89 picometers) and atoms in silicon at 0.78 ångström (78 picometers) at magnifications of 50 million times (see DOE/BES). The ability to determine the positions of atoms within materials has made the HRTEM an indispensable tool for nano-technologies research and development in many fields, including heterogeneous catalysis and the development of semiconductor devices for electronics and photonics.
Transmission electron microscopes produce two-dimensional images.
Unlike the TEM, where electrons are detected by beam transmission, the Scanning Electron Microscope (SEM)[5] produces images by detecting secondary electrons which are emitted from the surface due to excitation by the primary electron beam. In the SEM, the electron beam is rastered across the sample, with detectors building up an image by mapping the detected signals with beam position.
Generally, the TEM resolution is about an order of magnitude better than the SEM resolution, however, because the SEM image relies on surface processes rather than transmission it is able to image bulk samples and has a much greater depth of view, and so can produce images that are a good representation of the 3D structure of the sample.
In addition there is a Reflection Electron Microscope (REM). Like TEM, this technique involves electron beams incident on a surface, but instead of using the transmission (TEM) or secondary electrons (SEM), the reflected beam is detected. This technique is typically coupled with Reflection High Energy Electron Diffraction and Reflection high-energy loss spectrum (RHELS). Another variation is Spin-Polarized Low-Energy Electron Microscopy (SPLEEM), which is used for looking at the microstructure of magnetic domains [6].
main article: Scanning Transmission Electron Microscopy STEM
Materials to be viewed under an electron microscope may require processing to produce a suitable sample. The technique required varies depending on the specimen and the analysis required:
Electron microscopes are expensive to buy and maintain. They are dynamic rather than static in their operation: requiring extremely stable high-voltage supplies, extremely stable currents to each electromagnetic coil/lens, continuously-pumped high-/ultra-high-vacuum systems, and a cooling water supply circulation through the lenses and pumps. As they are very sensitive to vibration and external magnetic fields, microscopes aimed at achieving high resolutions must be housed in buildings (sometimes underground) with special services. Newer generations of TEM operating at lower voltages (around 5 kV) do not have stringent voltage supply, lens coil current, cooling water or vibration isolation requirements and as such are much less expensive to buy and far easier to install and maintain.
The samples have to be viewed in vacuum, as the molecules that make up air would scatter the electrons. Recent advances have allowed hydrated samples to be imaged using an environmental scanning electron microscope.
Scanning electron microscopes usually image conductive or semi-conductive materials best. Non-conductive materials can be imaged by an environmental scanning electron microscope. A common preparation technique is to coat the sample with a several-nanometer layer of conductive material, such as gold, from a sputtering machine; however this process has the potential to disturb delicate samples.
The samples have to be prepared in many ways to give proper detail, which may result in artifacts purely the result of treatment. This gives the problem of distinguishing artifacts from material, particularly in biological samples. Scientists maintain that the results from various preparation techniques have been compared, and as there is no reason that they should all produce similar artifacts, it is therefore reasonable to believe that electron microscopy features correlate with living cells. In addition, higher-resolution work has been directly compared to results from X-ray crystallography, providing independent confirmation of the validity of this technique. Recent work performed on unfixated, vitrified specimens has also been performed, further confirming the validity of this technique.