The Scanning Electron Microscope (SEM) produces images by probing the specimen with a focused electron beam that is scanned across a rectangular area of the specimen (raster scanning).
There are two families of electron guns:
Field emission gun (FEG) is used to produce an electron beam that is smaller in diameter, more coherent and up to three orders of magnitude greater current density or brightness.
|Emission Current (µA)
Energy of electrons is depending of Voltage: 1 Kev to 50KeV
Current (A): Number of electrons /unit of time
1 amp = 1 coulomb/sec 1 coulomb ~ 6 x1018 electrons
Example if the current measured at sample is around 10-9A to 10-12 A then the number of electrons is around 6X106 to 6X109 electrons/sec.
ESEM is a variety of SEM called environmental scanning electron microscope. It can produce images of sufficient quality and resolution with the samples being wet or contained in low vacuum or gas. This greatly facilitates imaging biological samples that are unstable in the high vacuum of conventional electron microscopes. The major disadvantage of transmission electron microscope is the need for extremely thin sections of the specimens, typically about 100 nanometers. Biological specimens are typically required to be chemically fixed, dehydrated and embedded in a polymer resin to stabilize them sufficiently to allow ultrathin sectioning. Sections of biological specimens, organic polymers and similar materials may require special treatment with heavy atom labels in order to achieve the required image contrast.
ESEM is especially useful for non-metallic, uncoated and biological materials. The presence of gas, mainly Argon, around a sample permits to work with pressure greater than 500 Pa compared to conventional SEM requirements samples under vacuum about 10-3 to 10-4 Pa. This vacuum level creates the possibility to operate on non-conductive samples without any preparation or hydrated specimens without charging.
In a Transmission Electron Microscope (TEM), the electron beam is accelerated by an anode typically at +100 keV (40 to 400 keV) with respect to the cathode, focused by electrostatic and electromagnetic lenses, and transmitted through the specimen that is in part transparent to electrons and in part scatters them out of the beam. When it emerges from the specimen, the electron beam carries information about the structure of the specimen that is magnified by the objective lens system of the microscope.
The spatial variation in this information (the “image”) may be viewed by projecting the magnified electron image onto a fluorescent viewing screen coated with a phosphor or scintillator material such as zinc sulfide. Alternatively, the image can be photographically recorded by exposing a photographic film or plate directly to the electron beam, or a high-resolution phosphor may be coupled by means of a lens optical system or a fiber optic light-guide to the sensor of a digital camera. The image detected by the digital camera may be displayed on a monitor or computer.
A transmission electron microscope can achieve better than 50 pm resolution and magnifications of up to about 10,000,000x whereas most light microscopes are limited by diffraction to about 200 nm resolution and useful magnifications below 2000x. Generally, the image resolution of an SEM is at least an order of magnitude poorer than that of a TEM. However, because the SEM image relies on surface processes rather than transmission, it is able to image bulk samples up to many centimeters in size and (depending on instrument design and settings) has a great depth of field, and so can produce images that are good representations of the three dimensional shape of the sample.
The Scanning Transmission Electron Microscope (STEM) rasters a focused incident probe across a specimen that (as with the TEM) has been thinned to facilitate detection of electrons scattered through the specimen. The high resolution of the TEM is thus possible in STEM. The focusing action (and aberrations) occurs before the electrons hit the specimen in the STEM, but afterward in the TEM.
Focused ion beam, also known as FIB, is a technique used particularly in the semiconductor industry, materials science and increasingly in the biological field for site-specific analysis, deposition, and ablation of materials. A FIB setup is a scientific instrument that resembles a scanning electron microscope (SEM). However, while the SEM uses a focused beam of electrons to image the sample in the chamber, a FIB setup uses a focused beam of ions instead. Unlike an electron microscope, FIB is inherently destructive to the specimen.
When the high-energy gallium ions strike the sample, they will sputter atoms from the surface. Gallium atoms will also be implanted into the top few nanometers of the surface, and the surface will be made amorphous. A FIB-SEM consists in a system with both electron and ion beam columns, allowing the same feature to be investigated using either of the beams. A FIB-SEM system uses a beam of Ga+ ion to mill into the surface to locate a feature or defect of interest. The integrated SEM then uses a focused beam of electrons to image the sample in the chamber.