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.
The addition of an aberration corrector to electron microscopes enables electron probes with sub-angstrom 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 (LVEM)
The low-voltage electron microscope (LVEM) is a combination of SEM, TEM and STEM in one instrument, which operated at relatively low electron accelerating voltage of 5 kV. 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. Sectioned samples generally need to be thinner than they would be for conventional STEM (20–70 nm). Resolutions of a few nm are possible in TEM, SEM and STEM modes.
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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