Ferrer, Lara Melissa V.

3 ChE A

Date submitted: Aug. 15, 2011

TRANSMISSION ELECTRON MICROSCOPE (TEM)

I.

INTRODUCTION Transmission electron microscopy (TEM) is a microscopy technique whereby a

beam of electrons is transmitted through an ultra thin specimen, interacting with the specimen as it passes through. An image is formed from the interaction of the electrons transmitted through the specimen; the image is magnified and focused onto an imaging device, such as a fluorescent screen, on a layer of photographic film, or to be detected by a sensor such as a CCD camera. TEMs are capable of imaging at a significantly higher resolution than light microscopes, owing to the small de Broglie wavelength of electrons. This enables the instrument's user to examine fine detaileven as small as a single column of atoms, which is tens of thousands times smaller than the smallest resolvable object in a light microscope. TEM forms a major analysis method in a range of scientific fields, in both physical and biological sciences. TEMs find application in cancer research, virology, materials science as well as pollution, nanotechnology, and semiconductor research. At smaller magnifications TEM image contrast is due to absorption of electrons in the material, due to the thickness and composition of the material. At higher magnifications complex wave interactions modulate the intensity of the image, requiring expert analysis of observed images. Alternate modes of use allow for the TEM to observe modulations in chemical identity, crystal orientation, electronic structure and sample induced electron phase shift as well as the regular absorption based imaging. The first TEM was built by Max Knoll and Ernst Ruska in 1931, with this group developing the first TEM with resolving power greater than that of light in 1933 and the first commercial TEM in 1939.[1] TEMs provided a means to go beyond the magnification and resolution limits of light microscopes, allowing for magnification of up to 100,000x and resolutions in the nanometer range.[2] It has been used in all areas of biological and biomedical investigations because of its ability to view the finest cell structures. It is also used as a

diagnostic tool in hospital pathology labs. For the crystallographer, metallurgist or semiconductor research scientist, current high voltage/high resolution TEMs, utilizing 200 keV to 1 MeV, have permitted the routine imaging of atoms, allowing materials researchers to monitor and design materials with custom-tailored properties. With the addition of energy dispersive X-ray analysis (EDXA) or energy loss spectrometry (EELS), the TEM can also be used as an elemental analysis tool, capable of identifying the elements in areas less than 0.5m in diameter.[3]

II.
A.

PRINCIPLES & CONCEPTS

How a Transmission Electron Microscope Works

An old-fashioned slide projector works by projecting light through a film layer. As the light passes through the film, it interacts with the film and specific areas of the film lets light pass through unobstructed, other areas absorb light and doesn't let it pass through, and still some absorb part of the light and lets only a fraction get through. The light that does go through is hits the lenses found on the other side and the resulting image is projected onto a screen. The TEM works similarly. In the case of the TEM, though, a beam of electrons are focused on a single, pinpoint spot or element on the sample being studied. The electrons interact with the sample and only those that go past unobstructed hit the phosphor screen on the other side. At this point, the electrons are converted to light and an image is formed. The dark areas of the image correspond to areas on the specimen where fewer electrons were able to pass through (either absorbed or scattered upon impact); the lighter areas are where more electrons did pass through, although the varying amounts of electrons in these areas enable the user to see structures and gradients. The 'lenses' in a TEM are not the same as lenses in a conventional microscope; these are actually EM devices that can 'focus' the electron beam to the desired wavelength or size. In much the same way as a light microscope, however, the amount of power used to generate electrons allows for higher magnification or better resolutions.

B.

Principles of Operation

The transmission electron microscope uses a high energy electron beam transmitted through a very thin sample to image and analyze the microstructure of materials with atomic scale resolution. The electrons are focused with electromagnetic lenses and the image is observed on a fluorescent screen, or recorded on film or digital camera. The electrons are accelerated at several hundred kV, giving wavelengths much smaller than that of light: 200kV electrons have a wavelength of 0.025. However, whereas the resolution of the optical microscope is limited by the wavelength of light, that of the electron microscope is limited by aberrations inherent in electromagnetic lenses, to about 1-2 . Because even for very thin samples one is looking through many atoms, one does not usually see individual atoms. Rather the high resolution imaging mode of the microscope images the crystal lattice of a material as an interference pattern between the transmitted and diffracted beams. This allows one to observe planar and line defects, grain boundaries, interfaces, etc. with atomic scale resolution. The brightfield/darkfield imaging modes of the microscope, which operate at intermediate magnification, combined with electron diffraction, are also invaluable for giving information about the morphology, crystal phases, and defects in a material. Finally the microscope is equipped with a special imaging lens allowing for the observation of micromagnetic domain structures in a field-free environment. The TEM is also capable of forming a focused electron probe, as small as 20 , which can be positioned on very fine features in the sample for microdiffraction information or analysis of x-rays for compositional information. The latter is the same signal as that used for EMPA ( Electron Probe Xray Microanalyzer) and SEM (Scanning Electron Microscope) composition analysis, where the resolution is on the order of one micron due to beam spreading in the bulk sample. The spatial resolution for this compositional analysis in TEM is much higher, on the order of the probe size, because the sample is so thin. Conversely the signal is much smaller and therefore less quantitative. The high brightness field-emission gun improves the sensitivity and resolution of x-ray compositional analysis over that available with more traditional thermionic sources.

C.

Restrictions on Samples

Sample preparation for TEM generally requires more time and experience than for most other characterization techniques. A TEM specimen must be approximately 1000 or less in thickness in the area of interest. The entire specimen must fit into a 3mm diameter cup and be less than about 100 microns in thickness. A thin, disc shaped sample with a hole in the middle, the edges of the hole being thin enough for TEM viewing, is typical. The initial disk is usually formed by cutting and grinding from bulk or thin film/substrate material, and the final thinning done by ion milling. Other specimen preparation possibilities include direct deposition onto a TEM-thin substrate (Si3N4, carbon); direct dispersion of powders on such a substrate; grinding and polishing using special devices (t-tool, tripod); chemical etching and electropolishing; lithographic patterning of walls and pillars for cross-section viewing; and focused ion beam (FIB) sectioning for site specific samples. Artifacts are common in TEM samples, due both to the thinning process and to changing the form of the original material. For example surface oxide films may be introduced during ion milling and the strain state of a thin film may change if the substrate is removed. Most artifacts can either be minimized by appropriate preparation techniques or be systematically identified and separated from real information. D. Training and Service

TEM training is available on an as-needed basis. Basic training for inexperienced TEM users requires a minimum of three four-hour sessions which can be done in groups of two or three. Additional training in specific TEM techniques is available on an asneeded basis following basic training. TEM users with prior experience will be trained at the level required. An accredited TEM laboratory course is periodically offered through the Materials Science and Engineering Department. Training in TEM specimen preparation is also available. Specimen prep for TEM requires more time and effort than for most other characterization techniques since a TEM specimen must be approximately 1000 or less in thickness in the area of interest and the entire specimen must fit into a shallow 3mm diameter cup. There are a variety of

techniques and instruments available for thinning materials to these specifications. Please refer to the TEM specimen preparation page for more details. TEM service is available for projects where extensive instrument training is not practical. However, users must generally make or provide their own TEM-ready samples.

III.

PARTS AND LABEL

1. High tension cable 2.

Electron emitter

3. Stepper motors for centering the

electron beam

4. Condenser 5. Aperture controls 6. Specimen holder 7. Objective lens 8. Projector lens 9. Optical binoculars 10. Fluorescent screen 11. Vacuum pump leads 12. Goniometer13. Vacuum and magnification control 14. Focusing control

IV.

SCHEMATIC DIAGRAM OF OPERATION

In the next page is a schematic diagram of a Transmission Electron Microscope. On the equivalent diagram for your particular make of microscope, find the most important lens, the objective. Work from there, find the specimen, the condenser lenses (somewhere above the objective), the diffraction and projector lenses (somewhere below

the objective). Find as many double deflection coils as you can: between the gun and condensers, the condensers and the specimen, and the diffraction lens and, possibly, within the projector lenses. Find all the aperture mechanisms. Below are the variables of a TEM:

The setting or current through each lens (about 7 variables) The setting of each double-deflection coil and their rocking points for both shift and tilt: for three sets, thats 4 settings for each x and y: a total of 24 settings. At least three sets of stigmators: another 6 variables. The physical height of the specimen, the physical aperture settings, and some physical alignments we havent worried about: another 10 or so variables.

V.

APPLICATIONS

The transmission electron microscope is used to characterize the microstructure of materials with very high spatial resolution. Information about the morphology, crystal structure and defects, crystal phases and composition, and magnetic microstructure can be obtained by a combination of electron-optical imaging (sub-ngstrom in the Titan, 2.5 point resolution in the Tecnai), electron diffraction, and small probe capabilities. Further, the Titan provides significant in situ capabilities, allowing for the investigation of how material structure can evolve due to different environmental factors. The trade-off for this diverse range of structural information and high resolution is the challenge of producing very thin samples for electron transmission. The TEM has its primary uses in metallurgy (or the study of metals and minerals) and the biological sciences, especially in the study of cells at the molecular level. TEMs have been particularly useful in metallurgy, especially in terms of developing images of crystals and metals at the molecular level allowing scientists to study their structure, interactions and identify flaws. The downside to the TEM lies in the specimens that can be studied these have to be 'sliced' very, very thinly to ensure that they are 'electron transparent'; they must also be placed in a vacuum. As such, preparation of specimens is often very time consuming and requires expert handling. This leads to concerns that specimens prepared for TEM study will be inadvertently damaged in the process. This raises the question of whether the specimen is as pure as can be expected. Finally, there are also concerns that the bombardment of electrons may damage the specimen under scrutiny especially if these are biological samples.

VI.

REFERENCES

1. "Configuration for the enlarged imaging of objects by electron beams". May 30, 1931. 2. Broglie, L. (1928). "La nouvelle dynamique des quanta".lectrons et Photons: Rapports et Discussions du Cinquime Conseil de Physique. Solvay. 3. Hawkes, P. (Ed.) (1985). The beginnings of Electron Microscopy. Academic Press. 4. Crewe, Albert V; Isaacson, M. and Johnson, D. (1969). "A Simple Scanning Electron Microscope". Rev. Sci. Inst. 40: 241 246.Bibcode 1969RScI...40..241C. doi:10.1063/1.1683910. 5. Crewe, Albert V; Wall, J. and Langmore, J. (1970). "Visibility of a single atom". Science 168 (3937): 8075. PMID 17731040. 6. Egerton, R (2005). Physical principles of electron microscopy. Springer. ISBN 0387258000. 1338 1340. Bibcode1970Sci...168.1338C. doi:10.1126/science.168.3937.1338.ISSN 0036-