When high-energy electrons reach the sample, several electronic and X-ray signals are generated. These include: Some scanning electron microscopes can reach a resolution of less than 1 nm. An account of the early days of scanning electron microscopy was presented by McMullan.   Although Max Knoll took a photograph with an object field width of 50 mm, which shows channel contrast using an electron beam scanner, it was Manfred von Ardenne who invented a high-resolution microscope in 1937 by scanning a very small grid with an enlarged and finely focused electron beam. Ardenne applied electron beam scanning to surpass the resolution of the transmission electron microscope (TEM) and mitigate the significant chromatic aberration problems inherent in real-world imaging at the TEM. He also discussed the different detection modes, the possibilities and theory of SEM, as well as the construction of the first high-resolution SEM.  Further work was reported by the Zworykin group, followed by the Cambridge groups in the 1950s and early 1960s. led to the commercialization of the Cambridge Scientific Instrument Company`s first commercial instrument under the name “stereoscan” in 1965. which was delivered to DuPont. Since SEM uses vacuum conditions and electrons to form an image, special preparations must be made on the sample.
All water must be removed from the samples, as the water would evaporate under vacuum. All metals are conductive and require no preparation before use. All nonmetals shall be made conductive by covering the sample with a thin layer of conductive material. This is done using a device called a “sputter coater”. Scanning electron microscopy works by scanning a sample with electron beams. An electron gun fires these beams, which then accelerate the scanning electron microscope column. Taking advantage of the SEM`s large field of view, the Everhart-Thornley detector (ETD) acquired a low-magnification image with a residence time of 30 μs, covering a field of view of several square millimeters (Fig. 17.9A).
Based on this reference image, a specific region was selected for focusing fluorescence and electron microscopes. To do this, the sample was translated to place the selected region in the center of the field of view and focusing procedures were performed. When electrons in the microscope interact with a sample, it can create different types of other electrons, photons, and irradiation. Hairpin tungsten cannon that generates a high-diameter electron beam to form high-resolution images. The two types of electrons essential for imaging are backscattered electrons (BSE) and secondary electrons (ES). The capacitor lens controls the size of the beam and determines the number of electrons in the beam. The size of the beam determines the resolution of the image. The wavelength is therefore a limiting factor in the resolution of optical microscopes.
Electron microscopes overcome this, as the shorter wavelengths of electrons allow for better resolution. The position of the electron beam on the sample is controlled by scanning coils located above the lens of the objective. These coils allow the beam to be scanned over the surface of the sample. This beam grid or scan is used to collect information about an area defined on the sample. As a result of the electron-sample interaction, a series of signals are generated. These signals are then detected by appropriate detectors. The SEM is not a camera and the detector does not continuously form images like a CCD or film. Unlike an optical system, the resolution is not limited by the diffraction limit, the fineness of the lenses or mirrors, or the resolution of the detector array. The focusing optics can be large and coarse, and the SE detector is the size of a fist and easily detects current. Instead, the spatial resolution of SEM depends on the size of the electron point, which in turn depends on both the wavelength of the electrons and the electron-optical system that produces the raster beam.
Resolution is also limited by the size of the interaction volume, the volume of sample material interacting with the electron beam. The size of the point and the volume of interaction are both important in relation to the distances between atoms, so the resolution of the SEM is not high enough to image individual atoms, as is possible with a transmission electron microscope (TEM). However, SEM has offsetting advantages, including the ability to image a relatively large area of the sample; the ability to image bulk solids (not just thin films or sheets); and the variety of analytical modes available to measure sample composition and properties. Depending on the instrument, the resolution can be between less than 1 nm and 20 nm. Since 2009, the world`s highest resolution conventional SEM (≤30 kV) has been capable of achieving a point resolution of 0.4 nm with a secondary electron detector.  The electron beam, which generally has an energy between 0.2 keV and 40 keV, is focused by one or two capacitor lenses at a point about 0.4 nm to 5 nm in diameter. The beam passes through pairs of scanning coils or pairs of deflector plates in the electron column, usually in the final lens, which deflects the beam into the x and y axes so that it sweeps like a raster over a rectangular area of the sample surface. If the SEM is equipped with a cold stage for cryomicroscopy, cryofixation can be used and low-temperature scanning electron microscopy can be performed on cryogenically fixed samples.
 Cryofixed samples can be cryobroken under vacuum in a special device to expose the internal structure, coated with sputtering and transferred to the SEM cryotable while still frozen.  Low temperature scanning electron microscopy (LT-SEM) is also applicable to imaging temperature-sensitive materials such as ice and greases.  SEM stands for scanning electron microscope. SEM is a microscope that uses electrons instead of light to form an image. Since their development in the early 1950s, scanning electron microscopes have developed new areas of research in the medical and physical sciences. SEM allowed researchers to study a wider variety of samples. SEM and ESEM are particularly suitable for detecting the surface and texture characteristics of large particulate units (approximately >1 μm) and for rough estimation of particle size distribution. Qualitative 3D morphological information can be easily extracted from the SEM, which operates in secondary electron mode or backscattered electron mode, and the extent of aggregation between particles (assuming it is not an artifact generated by overloaded sample strains) can be easily documented with a resolution of up to ∼50 nm. Due to its ease of use, SEM should be selected for routine analysis of samples, especially atmospheric and soil particles. Preliminary qualitative surveys should help the operator to focus either on general trends (e.g.
the main classes of particle types, sizes or associations) or on significant specimens of the sample (e.g. characteristic aggregation between two particle types or prevalence of a narrow size class for a given feature type). The adaptability of scanning electron microscopes makes them ideal for a wide range of scientific, research, industrial and commercial applications. The electron source generates electrons at the end of the microscope column. The dry sample is usually mounted on a sample strip with an adhesive such as epoxy resin or electrically conductive double-sided tape and coated with gold or a gold/palladium alloy before examination under a microscope. Samples can be cut (with a microtome) if information about the internal ultrastructure of the organism needs to be exposed for imaging. The spray coating uses an electric field and argon gas. The sample is placed in a small chamber located under vacuum. Argon gas and an electric field cause an electron to withdraw from the argon, causing the atoms to charge positively. The argon ions are then attracted to a negatively charged gold leaf. Argon ions cause gold atoms to fall off the surface of the gold leaf. These gold atoms fall and settle on the surface of the sample, creating a thin coating of gold.
Scanning electron microscopes (REM) use an electron beam to image samples with resolution up to the nanometer. The electrons are emitted by a filament and collimated into a beam in the electron source. The beam is then focused on the surface of the sample through a series of lenses in the electron column. How does an electronic lens work? And what kind of lenses are there? How are lenses combined in an electron column? In this blog, we will answer these questions and give a general overview of the functional principle of an electron column. The scanning electron microscope (SEM) is a microscope that uses electrons instead of light to form an image. There are many advantages to using SEM instead of an optical microscope. The electron beams scan the sample in a grid and scan the surface in lines from side to side, up and down. The scanning electron microscope (SEM) is one of the most widely used instrumental methods for the investigation and analysis of micro- and nanoparticle imaging characterization of solid objects.
One of the reasons why SEM is preferred for particle size analysis is its resolution of 10 nm, or 100 Å. Advanced versions of these instruments can achieve a resolution of about 2.5 nm (25 Å) (Goldstein, 2012). This instrument can also be used in conjunction with other related energy-dispersive X-ray microanalysis techniques (EDX, EDS, EDAX) to determine the composition or orientation of individual crystals or features.