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Electron Microscopy

The electron microscope is a very powerful microscope which can see things that normal microscopes cannot. There are 2 types of electron microscope: the transmission electron microscope and the scanning electron microscope.

The sample must be in a vacuum so that no air bubbles are on the produced image and also because the electrons are absorbed by the molecules in the air, this means that the electron microscope cannot be used to look at living cells. The tissue is soaked in alcohol to dehydrate it and must be stained in order to improve the contrast on the image produced called an electronmicrograph. To see the objects you need to use radiation with a shorter wavelength, this is done by getting metal very hot so that some of its electrons gain enough energy to escape from their orbits. The escaped electrons behave like electromagnetic radiation but have a very short wavelength. Electrons are negatively charged the microscope uses electromagnets to focus the image on to a fluorescent screen. Increasing the magnification in electron microscopy will result in an increase in the amount of visible detail. So the resolving power of the microscope depends on the wavelength of the electromagnetic radiation. The Transmission Electron Microscope The transmission electron microscope is used to look inside cells and allows us to see separate structures, particles which are as close together as 2 nanometres (the resolution). Increasing the magnification in electron microscopy will result in an increase in the amount of visible detail Electrons must be able to pass through the specimen and this means that part of the specimen appears bright but other parts absorb the electrons so they are electron-dense areas and therefore appear dark on the image. A problem with the microscope is that the cells must be cut into very thin sections using a knife made of glass. The cut tissue has to be about the thickness of the film around a soap bubble. Another problem is that the treatment of the tissue may introduce artefacts which may appear on the finished image but are not part of the natural specimen. This means that it is not always easy to be sure that what we see on a electronmicrograph really exists in that form. The microscope can be used at low as well as high magnifications and an image is formed from electrons emitted or reflected from the surface of a complete specimen. The Scanning Electron Microscope The scanning electron microscope gives a three dimensional effect and shows the surface of the specimen in detail. The scanning electron microscope scans electron beams to and fro across the surface of a complete specimen creating a 3-D image. The

The scanning electron microscope does not have such a high resolving power as the transmission electron microscope (20 nanometres for the scanning microscope compared to 2 nanometres for the transmission microscope) but this is still better than a light microscope.specimen does not have to be cut into thin sections like it needs to be with the transmission microscope but the surface is coated with a thin film of gold. . Apart from the specimens not needing to be as thin the scanning microscope has the same limitations as the transmission electron microscope such as the specimen needing to be in a vacuum and the problems with artefacts.