This article includes a list of general references, but it lacks sufficient corresponding inline citations .(January 2015) |
In situ electron microscopy is an investigatory technique where an electron microscope is used to watch a sample's response to a stimulus in real time. Due to the nature of the high-energy beam of electrons used to image a sample in an electron microscope, microscopists have long observed that specimens are routinely changed or damaged by the electron beam. Starting in the 1960s, and using transmission electron microscopes (TEMs), scientists made deliberate attempts to modify materials while the sample was in the specimen chamber, and to capture images through time of the induced damages.
Also in the 1960s, materials scientists using TEMs began to study the response of electron-transparent metal samples to irradiation by the electron beam. This was in order to understand more about metal fatigue during aviation and space flight. The experiments were performed on instruments with high accelerating voltages; the image resolution was low compared to the sub-nanometer resolution available with modern TEMs.
Improvements in electron microscopy from the 1960s onwards focused on increasing the spatial resolution. This required increased stability for the entire imaging platform, but particularly for the area around the specimen stage. Improved image-capture systems using charge-coupled device cameras and advances in specimen stages coupled with the higher resolution led to creating systems devoted to applying stimuli to samples in specialized holders, and capturing multiple frames or videos of the samples' responses.
In addition to materials samples, in situ electron microscopy is performed on biological specimens, and is used to conduct experiments involving mechanical, chemical, thermal, and electrical responses. Early experiments mostly used TEMs, because the image is captured in a single frame, whereas the scanning electron microscope must move or scan across the sample while the stimuli is being applied, altering the sample.
Early problems that limited in situ electron microscopy included mechanical vibration at all scales (from the microscope itself to the sample), and thermal and electrical interference, particularly at the specimen holder. These problems all required fast capture times. However a fast capture time creates an image with a low signal-to-noise ratio, limits the resolution of the image, and also limits the amount of time available for conducting the experiment. [1]
An electron microscope is a microscope that uses a beam of electrons as a source of illumination. They use electron optics that are analogous to the glass lenses of an optical light microscope to control the electron beam, for instance focusing them to produce magnified images or electron diffraction patterns. As the wavelength of an electron can be up to 100,000 times smaller than that of visible light, electron microscopes have a much higher resolution of about 0.1 nm, which compares to about 200 nm for light microscopes. Electron microscope may refer to:
A microscope is a laboratory instrument used to examine objects that are too small to be seen by the naked eye. Microscopy is the science of investigating small objects and structures using a microscope. Microscopic means being invisible to the eye unless aided by a microscope.
A scanning electron microscope (SEM) is a type of electron microscope that produces images of a sample by scanning the surface with a focused beam of electrons. The electrons interact with atoms in the sample, producing various signals that contain information about the surface topography and composition of the sample. The electron beam is scanned in a raster scan pattern, and the position of the beam is combined with the intensity of the detected signal to produce an image. In the most common SEM mode, secondary electrons emitted by atoms excited by the electron beam are detected using a secondary electron detector. The number of secondary electrons that can be detected, and thus the signal intensity, depends, among other things, on specimen topography. Some SEMs can achieve resolutions better than 1 nanometer.
Transmission electron microscopy (TEM) is a microscopy technique in which a beam of electrons is transmitted through a specimen to form an image. The specimen is most often an ultrathin section less than 100 nm thick or a suspension on a grid. An image is formed from the interaction of the electrons with the sample as the beam is transmitted through the specimen. The image is then magnified and focused onto an imaging device, such as a fluorescent screen, a layer of photographic film, or a detector such as a scintillator attached to a charge-coupled device or a direct electron detector.
Atomic force microscopy (AFM) or scanning force microscopy (SFM) is a very-high-resolution type of scanning probe microscopy (SPM), with demonstrated resolution on the order of fractions of a nanometer, more than 1000 times better than the optical diffraction limit.
Photoemission electron microscopy is a type of electron microscopy that utilizes local variations in electron emission to generate image contrast. The excitation is usually produced by ultraviolet light, synchrotron radiation or X-ray sources. PEEM measures the coefficient indirectly by collecting the emitted secondary electrons generated in the electron cascade that follows the creation of the primary core hole in the absorption process. PEEM is a surface sensitive technique because the emitted electrons originate from a shallow layer. In physics, this technique is referred to as PEEM, which goes together naturally with low-energy electron diffraction (LEED), and low-energy electron microscopy (LEEM). In biology, it is called photoelectron microscopy (PEM), which fits with photoelectron spectroscopy (PES), transmission electron microscopy (TEM), and scanning electron microscopy (SEM).
An X-ray microscope uses electromagnetic radiation in the X-ray band to produce magnified images of objects. Since X-rays penetrate most objects, there is no need to specially prepare them for X-ray microscopy observations.
A scanning transmission electron microscope (STEM) is a type of transmission electron microscope (TEM). Pronunciation is [stɛm] or [ɛsti:i:ɛm]. As with a conventional transmission electron microscope (CTEM), images are formed by electrons passing through a sufficiently thin specimen. However, unlike CTEM, in STEM the electron beam is focused to a fine spot which is then scanned over the sample in a raster illumination system constructed so that the sample is illuminated at each point with the beam parallel to the optical axis. The rastering of the beam across the sample makes STEM suitable for analytical techniques such as Z-contrast annular dark-field imaging, and spectroscopic mapping by energy dispersive X-ray (EDX) spectroscopy, or electron energy loss spectroscopy (EELS). These signals can be obtained simultaneously, allowing direct correlation of images and spectroscopic data.
Transmission electron cryomicroscopy (CryoTEM), commonly known as cryo-EM, is a form of cryogenic electron microscopy, more specifically a type of transmission electron microscopy (TEM) where the sample is studied at cryogenic temperatures. Cryo-EM, specifically 3-dimensional electron microscopy (3DEM), is gaining popularity in structural biology.
Metallography is the study of the physical structure and components of metals, by using microscopy.
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. FIB can also be incorporated in a system with both electron and ion beam columns, allowing the same feature to be investigated using either of the beams. FIB should not be confused with using a beam of focused ions for direct write lithography. These are generally quite different systems where the material is modified by other mechanisms.
High-resolution transmission electron microscopy is an imaging mode of specialized transmission electron microscopes that allows for direct imaging of the atomic structure of samples. It is a powerful tool to study properties of materials on the atomic scale, such as semiconductors, metals, nanoparticles and sp2-bonded carbon. While this term is often also used to refer to high resolution scanning transmission electron microscopy, mostly in high angle annular dark field mode, this article describes mainly the imaging of an object by recording the two-dimensional spatial wave amplitude distribution in the image plane, similar to a "classic" light microscope. For disambiguation, the technique is also often referred to as phase contrast transmission electron microscopy, although this term is less appropriate. At present, the highest point resolution realised in high resolution transmission electron microscopy is around 0.5 ångströms (0.050 nm). At these small scales, individual atoms of a crystal and defects can be resolved. For 3-dimensional crystals, it is necessary to combine several views, taken from different angles, into a 3D map. This technique is called electron tomography.
A scanning helium ion microscope is an imaging technology based on a scanning helium ion beam. Similar to other focused ion beam techniques, it allows to combine milling and cutting of samples with their observation at sub-nanometer resolution.
Electron-beam-induced deposition (EBID) is a process of decomposing gaseous molecules by an electron beam leading to deposition of non-volatile fragments onto a nearby substrate. The electron beam is usually provided by a scanning electron microscope, which results in high spatial accuracy and the possibility to produce free-standing, three-dimensional structures.
The environmental scanning electron microscope (ESEM) is a scanning electron microscope (SEM) that allows for the option of collecting electron micrographs of specimens that are wet, uncoated, or both by allowing for a gaseous environment in the specimen chamber. Although there were earlier successes at viewing wet specimens in internal chambers in modified SEMs, the ESEM with its specialized electron detectors and its differential pumping systems, to allow for the transfer of the electron beam from the high vacuum in the gun area to the high pressure attainable in its specimen chamber, make it a complete and unique instrument designed for the purpose of imaging specimens in their natural state. The instrument was designed originally by Gerasimos Danilatos while working at the University of New South Wales.
Alex K. Zettl is an American experimental physicist, educator, and inventor.
A Low-voltage electron microscope (LVEM) is an electron microscope which operates at accelerating voltages of a few kiloelectronvolts (keV) or less. Traditional electron microscopes use accelerating voltages in the range of 10-1000 keV.
Scanning transmission X-ray microscopy (STXM) is a type of X-ray microscopy in which a zone plate focuses an X-ray beam onto a small spot, a sample is scanned in the focal plane of the zone plate and the transmitted X-ray intensity is recorded as a function of the sample position. A stroboscopic scheme is used where the excitation is the pump and the synchrotron X-ray flashes are the probe. X-ray microscopes work by exposing a film or charged coupled device detector to detect X-rays that pass through the specimen. The image formed is of a thin section of specimen. Newer X-ray microscopes use X-ray absorption spectroscopy to heterogeneous materials at high spatial resolution. The essence of the technique is a combination of spectromicroscopy, imaging with spectral sensitivity, and microspectroscopy, recording spectra from very small spots.
Liquid-phase electron microscopy refers to a class of methods for imaging specimens in liquid with nanometer spatial resolution using electron microscopy. LP-EM overcomes the key limitation of electron microscopy: since the electron optics requires a high vacuum, the sample must be stable in a vacuum environment. Many types of specimens relevant to biology, materials science, chemistry, geology, and physics, however, change their properties when placed in a vacuum.
Transmission Kikuchi Diffraction (TKD), also sometimes called transmission-electron backscatter diffraction (t-EBSD), is a method for orientation mapping at the nanoscale. It’s used for analysing the microstructures of thin transmission electron microscopy (TEM) specimens in the scanning electron microscope (SEM). This technique has been widely utilised in the characterization of nano-crystalline materials, including oxides, superconductors, and metallic alloys.