Electron Microscope
Submitted By:
Umair Faheem, 2003-ag-32 [Link]. (Hons.) Agri. Entomology.
Submitted to:
Dr. Shafqat Saeed, Assistant Professor, Agri. Entomology.
Electron Microscopy
What are Electron Microscopes?
Electron Microscopes are scientific instruments that use a beam of highly energetic electrons to examine objects on a very fine scale. This examination can yield the following information: Topography The surface features of an object or "how it looks", its texture; direct relation between these features and materials properties (hardness, reflectivity...etc.) Morphology The shape and size of the particles making up the object; direct relation between these structures and materials properties (ductility, strength, reactivity...etc.) Composition The elements and compounds that the object is composed of and the relative amounts of them; direct relationship between composition and materials properties (melting point, reactivity, hardness...etc.) Crystallographic Information How the atoms are arranged in the object; direct relation between these arrangements and materials properties (conductivity, electrical properties, strength...etc.)
Where did Electron Microscopes Come From?
Electron Microscopes were developed due to the limitations of Light Microscopes which are limited by the physics of light to 500x or 1000x magnification and a resolution of 0.2 micrometers. In the early 1930's this theoretical limit had been reached and there was a scientific desire to see the fine details not of the interior possible structures of organic Light cells (nucleus, mitochondria...etc.). This required 10,000x plus magnification which was just using Microscopes. The Transmission Electron Microscope (TEM) was the first type of Electron Microscope to be developed and is patterned exactly on the Light Transmission Microscope except that a focused beam of electrons is used instead of light to "see through" the specimen. It was developed by Max Knoll and Ernst Ruska in Germany in 1931. The first Scanning Electron Microscope (SEM) debuted in 1942 with the first commercial instruments around 1965. Its late development was due to the electronics involved in "scanning" the beam of electrons across the sample. An excellent article was just published in Scanning detailing the history of SEMs and I would encourage those interested to read it.
How do Electron Microscopes Work?
Electron Microscopes (EMs) function exactly as their optical counterparts except that they use a focused beam of electrons instead of light to "image" the specimen and gain information as to its structure and composition.
The basic steps involved in all EMs:
1. A stream of electrons is formed (by the Electron Source) and accelerated toward the specimen using a positive electrical potential 2. This stream is confined and focused using metal apertures and magnetic lenses into a thin, focused, monochromatic beam. 3. This beam is focused onto the sample using a magnetic lens 4. Interactions occur inside the irradiated sample, affecting the electron beam
These interactions and effects are detected and transformed into an image The above steps are carried out in all EMs regardless of type. A more specific treatment of the workings of two different types of EMs are described in more detail:
Transmission Electron Microscope Scanning Electron Microscope
Transmission Electron Microscope (TEM)
TEM works much like a slide projector. A projector shines a beam of light through (transmits) the slide, as the light passes through it is affected by the structures and objects on the slide. These effects result in only certain parts of the light beam being transmitted through certain parts of the slide. This transmitted beam is then projected onto the viewing screen, forming an enlarged image of the slide.
TEMs work the same way except that they shine a beam of electrons (like the light) through the specimen(like the slide). Whatever part is transmitted is projected onto a phosphor screen for the user to see. A more technical explanation of a typical TEMs workings is as follows (refer to the diagram below): 1. The "Virtual Source" at the top represents the electron gun, producing a stream of monochromatic electrons. 2. This stream is focused to a small, thin, coherent beam by the use of condenser lenses 1 and 2. The first lens (usually controlled by the "spot size knob") largely determines the "spot size"; the general size range of the final spot that strikes the sample. The second lens(usually controlled by the "intensity or brightness knob" actually changes the size of the spot on the sample; changing it from a wide dispersed spot to a pinpoint beam. 3. The beam is restricted by the condenser aperture (usually user selectable), knocking out high angle electrons (those far from the optic axis, the dotted line down the center) 4. The beam strikes the specimen and parts of it are transmitted 5. This transmitted portion is focused by the objective lens into an image 6. Optional Objective and Selected Area metal apertures can restrict the beam; the Objective aperture enhancing contrast by blocking out high-angle diffracted electrons, the Selected Area aperture enabling the user to examine the periodic diffraction of electrons by ordered arrangements of atoms in the sample 7. The image is passed down the column through the intermediate and projector lenses, being enlarged all the way 8. The image strikes the phosphor image screen and light is generated, allowing the user to see the image. The darker areas of the image represent those areas of the sample that fewer electrons were transmitted through (they are thicker or denser). The lighter areas of the image represent those areas of the sample that more electrons were transmitted through (they are thinner or less dense)
Scanning Electron Microscope (SEM)
A detailed explanation of how a typical SEM functions follows (refer to the diagram below):
1. The "Virtual Source" at the top represents the electron gun, producing a stream of monochromatic electrons. 2. The stream is condensed by the first condenser lens (usually controlled by the "coarse probe current knob"). This lens is used to both form the beam and limit the amount of current in the beam. It works in conjunction with the condenser aperture to eliminate the high-angle electrons from the beam 3. The beam is then constricted by the condenser aperture (usually not user selectable), eliminating some high-angle electrons 4. The second condenser lens forms the electrons into a thin, tight, coherent beam and is usually controlled by the "fine probe current knob" 5. A user selectable objective aperture further eliminates high-angle electrons from the beam 6. A set of coils then "scan" or "sweep" the beam in a grid fashion (like a television), dwelling on points for a period of time determined by the scan speed (usually in the microsecond range) 7. The final lens, the Objective, focuses the scanning beam onto the part of the specimen desired. 8. When the beam strikes the sample (and dwells for a few microseconds) interactions occur inside the sample and are detected with various instruments
9. Before the beam moves to its next dwell point these instruments count the number of interactions and display a pixel on a CRT whose intensity is determined by this number (the more reactions the brighter the pixel). 10. This process is repeated until the grid scan is finished and then repeated, the entire pattern can be scanned 30 times per second.
Precautions:
1. For TEMs the object must be finely sliced so the electrons may pass through them 2. For SEMs no slicing of the object is required but the object must be dehydrated so the electrons does not deflect with water molecules and also the tube containing object should be vacuumed so that the electrons does not deflect with air molecules.