Scanning transmission electron microscopy
A scanning transmission electron microscope (STEM) is a type of transmission electron microscope (TEM). Pronunciation is [stem] or [esti:i:em]. As with any transmission illumination scheme, the electrons pass through a sufficiently thin specimen. However, STEM is distinguished from conventional transmission electron microscopes (CTEM) by focusing the electron beam into a narrow spot which is scanned over the sample in a raster.
The rastering of the beam across the sample makes these microscopes suitable for analysis techniques such as mapping by energy dispersive X-ray (EDX) spectroscopy, electron energy loss spectroscopy (EELS) and annular dark-field imaging (ADF). These signals can be obtained simultaneously, allowing direct correlation of image and quantitative data.
By using a STEM and a high-angle detector, it is possible to form atomic resolution images where the contrast is directly related to the atomic number (z-contrast image). The directly interpretable z-contrast image makes STEM imaging with a high-angle detector appealing. This is in contrast to the conventional high resolution electron microscopy technique, which uses phase-contrast, and therefore produces results which need interpretation by simulation.
Usually a STEM is a conventional transmission electron microscope equipped with additional scanning coils, detectors and needed circuitry; however, dedicated STEMs are also manufactured.
In 1925, Louis de Broglie first theorized the wave-like properties of an electron, with a wavelength substantially smaller than visible light. This would allow the use of electrons to image objects much smaller than the previous diffraction limit set by visible light. The first STEM was built in 1938 by Baron Manfred von Ardenne, working in Berlin for Siemens. However, at the time the results were inferior to those of transmission electron microscopy, and von Ardenne only spent two years working on the problem. The microscope was destroyed in an air raid in 1944, and von Ardenne did not return to his work after WWII.
The technique was not developed further until the 1970s, when Albert Crewe at the University of Chicago developed the field emission gun and added a high quality objective lens to create a modern STEM. He demonstrated the ability to image atoms using an annular dark field detector. Crewe and coworkers at the University of Chicago developed the cold field emission electron source and built a STEM able to visualize single heavy atoms on thin carbon substrates.
Aberration-corrected STEM was demonstrated with 1.9 angstrom resolution in 1997 and soon after in 2000 with roughly 1.36 angstrom resolution. Aberration-corrected STEM provided the added resolution and beam current critical to the implementation of atomic resolution chemical mapping with spectroscopic techniques.
The addition of an aberration corrector to electron microscopes enables electron probes with sub-ångström diameters to be used. This has made it possible to identify individual atoms columns with unprecedented clarity.
High resolution scanning transmission electron microscopes require exceptionally stable room environments. In order to obtain atomic resolution imaging the room must have a limited amount of room vibration, temperature fluctuations, electromagnetic waves, and acoustic waves.
The first application of this method to the imaging of biological molecules was demonstrated in 1971. The motivation for STEM imaging of biological samples is particularly to make use of dark-field microscopy, where the STEM is more efficient than a conventional TEM, allowing high contrast imaging of biological samples without requiring staining. The method has been widely used to solve a number of structural problems in molecular biology.
Low-voltage electron microscope
A low-voltage electron microscope (LVEM) is operated at relatively low electron accelerating voltage between 5-25 kV. Some of these can be a combination of SEM, TEM and STEM in a single compact instrument. Low voltage increases image contrast which is especially important for biological specimens. This increase in contrast significantly reduces, or even eliminates the need to stain. Resolutions of a few nm are possible in TEM, SEM and STEM modes. The low energy of the electron beam means that permanent magnets can be used as lenses and thus a miniature column that does not require cooling can be used.
Electron energy loss spectroscopy
Electron energy loss spectroscopy (EELS) as a STEM measurement technique made possible with the addition of an electron spectrometer. The high-energy convergent electron beam in STEM provides local information of the sample, even down to atomic dimensions. With the addition of EELS, elemental identification is possible and even additional capabilities of determining electronic structure or chemical bonding of atomic columns. The low-angle inelastically scattered electrons used in EELS compliments the high-angle scattered electrons in ADF images by allowing both signals to be acquired simultaneously. EELS is a technique popular to STEM microscopists.
- Electron beam induced deposition
- Electron diffraction
- Electron energy loss spectroscopy (EELS)
- Electron microscope
- Energy filtered transmission electron microscopy (EFTEM)
- High-resolution transmission electron microscopy (HRTEM)
- Low-voltage electron microscopy (LVEM)
- Scanning confocal electron microscopy
- Scanning electron microscope (SEM)
- Transmission Electron Aberration-corrected Microscope
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- von Ardenne, M (1938). "Das Elektronen-Rastermikroskop. Praktische Ausführung". Z Tech Phys 19: 407–416.
- D. McMullan, SEM 1928 - 1965
- Crewe, Albert V; Isaacson, M.; Johnson, D. (1969). "A Simple Scanning Electron Microscope". Rev. Sci. Inst. 40 (2): 241–246. Bibcode:1969RScI...40..241C. doi:10.1063/1.1683910.
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|last1=in Authors list (help)
- Dellby, Niklas; O.L. Krivanek; P.D. Nellist; P.E. Batson; A.R. Lupini (2001). "Progress in aberration-corrected scanning transmission electron microscopy". J. Electron Microsc. 50: 177.
- Wall, J.S., 1971 "A high resolution scanning electron microscope for the study of single biological molecules" PhD thesis, University of Chicago
- Wall JS, Hainfeld JF (1986). "Mass mapping with the scanning transmission electron microscope". Annu Rev Biophys Biophys Chem 15: 355–76. doi:10.1146/annurev.bb.15.060186.002035. PMID 3521658.
- Hainfeld JF, Wall JS (1988). "High resolution electron microscopy for structure and mapping". Basic Life Sci 46: 131–47. PMID 3066333.
- Wall JS, Simon MN (2001). "Scanning transmission electron microscopy of DNA-protein complexes". Methods Mol Biol 148: 589–601. doi:10.1385/1-59259-208-2:589. ISBN 1-59259-208-2. PMID 11357616.
- Nebesářová1, Jana; Vancová, Marie (2007). "How to Observe Small Biological Objects in Low-Voltage Electron Microscope". Microscopy and Microanalysis 13 (3): 248–249. doi:10.1017/S143192760708124X (inactive 2015-02-01).
- Drummy, Lawrence, F.; Yang, Junyan; Martin, David C. (2004). "Low-voltage electron microscopy of polymer and organic molecular thin films". Ultramicroscopy 99 (4): 247–256. doi:10.1016/j.ultramic.2004.01.011. PMID 15149719.
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