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Dharmapadmi Pradnya Kasilani - Resume 2 Nanomedicine 3 Ta 2022
Dharmapadmi Pradnya Kasilani - Resume 2 Nanomedicine 3 Ta 2022
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History of SEM
• In 1933, Electron Microscopy was invented by Ernst Ruska, a German
physicist. He was a
• Awarded the Nobel Prize for physics for his invention in 1986
• In 1938, First SEM debuted by Manfred Von Ardenne
• In 1965, Cambridge Scientific Instrument (UK) and JOEL (Japan) first
commercialized SEM individually
Introduction of SEM
The light microscope used photons while the electron microscope used
electrons. Photons are substituted with electrons and glass lenses are
substituted with electromagnetic and electrostatic lenses. The size of the light
microscope is more than 100nm (the size of a Mycoplasma) and less than 1mm
(the size of a frog egg). The size of an electron microscope is more than 0.1nm
(the size of atoms) and less than 100µm (the size of eukaryotic cells). Electron
microscopes are scientific instruments that use a beam of highly energetic
electrons to examine objects on a very fine scale.
Applications of SEM
• Topography is used to observe the surface features of the sample
• Morphology is used to observe the shape and size of the sample
• Composition is used to observe the elements that the sample is composed
of and their relative amount (%) – purity
• Crystallographic info is used to observe how atoms are arranged in the
sample
The SEM image shows a more detailed picture of the object than the
compound microscope image and TEM image. The TEM image shows a more
detailed image of the object than the Compound microscope image.
Instrumentation
The parts of SEM include:
• Electron cannon
• Electron gun consisting of a cathode and anode
• The condenser lens controls the number of electrons traveling down the
column
• The objective lens focuses the beam into a spot on the sample
• The deflection coil helps to deflect the electron beam
• SED attracts the secondary electrons
• Additional sensors detect backscattered electrons and x-rays
• Vacuum pumps system
• Operation panel with focus, alignment, and magnification tools and a
joystick for positioning
• Screen for menu and image display
• Cryo-unit to prepare frozen material before insertion in the observation
chamber in Cryo-SEM mode
Formation of an image
High-energy electrons shoot and the outgoing electron/x-rays analyze.
SEM changes the magnification of an image by reducing the size of the area
scanned by the scan coils. This form of image processing is only in gray. This
image can be colorized through the use of feature-detection software. This is
usually for aesthetic effects and adding a realistic effect.
Resolution
• Best resolution that can be obtained size of the electron spot on the surface
of the sample
➢ The introduction of FEG has dramatically improved the resolution
of SEMs
• The volume from which the signal electrons are formed defines the
resolution
➢ SE image has a higher resolution than a BSE image
• Scanning speed :
➢ A weak signal requires slow speed to improve the signal-to-noise
ratio
➢ When doing a slow scan drift in the electron beam can affect the
accuracy of the analysis
Sample preparation
1. Coating specimen
A coating specimen is used to increase the conductivity of the specimen
and to prevent the high voltage charge on the specimen. All metals are
conductive and require no preparation before being used. Meanwhile, non-
metals need to be made conductive by using a device called a "sputter
coater". These non-metals will coat with thin layers i.e. 20nm – 30nm of
conductive metal such as gold, gold-palladium alloy, platinum, osmium,
iridium, tungsten, chromium, and graphite. Appropriate size sample that
fits in the specimen chamber. Mounted rigidly on a specimen holder in
specimen stub. For imaging in the SEM, specimens must be electrically
conductive and electrically grounded.
2. Cleaning the surface of the specimen
Cleaning the specimen surface is very important because the surface
contains a lot of unwanted deposits, such as dust, mud, soil, etc.
3. Dehydrating the specimen
Dehydration of the specimen using water must be removed, air drying
causes collapse and shrinkage, this is usually achieved by replacing the
water in the cell with an organic solvent such as ethanol or acetone,
dehydration is carried out with a graded sequence of ethanol or acetone.
4. Drying the specimen
Dry the specimen by ensuring that the specimen is completely dry.
Otherwise, the sample will be destroyed.
5. Mounting the specimen
Furthermore, the way to place the specimen is that the specimen must be
mounted on a holder, rigidly mounted on specimen holder called a
specimen stub. For dry specimens, it is attached to the specimen stub
using an adhesive such as epoxy resin or electrically conductive double-
sided adhesive tape.
SEM Advantage
• Gives detailed 3D and topographical imaging and versatile information
garnered from different detectors.
• Works very fast.
• Modern SEMs allow for the generation of data in digital form.
• Most SEM samples require minimal preparation actions.
SEM Disadvantage
• SEMs are expensive and large.
• Special training is required to operate an SEM.
• The preparation of samples can result in artifacts.
• Limited to solid samples.
• Carry a small risk of radiation exposure associated with the electrons that
scatter from beneath the sample surface.