Dark field electron microscopy

Methods BF = bright field LM of tissue impression smears, wet-mounts, stained whole mounts LM = light microscopy PH = phase microscopy DF = dark-field microscopy TEM/SEM = transmission/scanning electron microscopy of sections or of purified or semi-purified virus ELISA = enzyme-linked immunosorbent assay PAb = polyclonal antibodies MAb = monoclonal antibodies DBH = dot blot hybridization ISH = in situ hybridization PCR = polymerase chain reaction RT-PCR = reverse transcription PCR. 

In dark-field electron microscopy it is not the transmitted beam which is used to construct an image but, rather, a beam diffracted from one facet of the object under investigation. One method for doing this is to shift the aperture of the microscope so that most of the beam is blocked and only those electrons... 

There are three primary image modes that are used in conventional TEM work, bright-field microscopy, dark-field microscopy, and high-resolution electron microscopy. In practice, the three image modes differ in the way in which an objective diaphragm is used as a filter in the back focal plane. 

S. J. Pennycook. EMSA Bulletin. 19, 67, 1989. A summary of compositional imaging using a high-angle annular dark-field detector in a field emission STEM instrument published by the Electron Microscopy Society of America, Box EMSA Woods Hole, MA 02543. 

The structure of the metal particles dispersed on a silica powder support ( Aerosil 380, 70 A average silica particle diameter) has been studied by Avery and Sanders (47) using electron microscopy in both bright and dark field, to determine the extent to which the metal particles were multiply twinned or of ideal structure. Platinum, palladium, and gold were examined. These catalysts were prepared by impregnation using an aqueous solution of metal halide derivatives, were dried at 100°-150°C, and were hydrogen... 

Dark field Visualization technique for ashes produced by microincineration and fluorescence microscopy useful for low-contrast subjects Electron systems imaging EM shadowing Detection, localization, and quantitation of light elements Structural information from ordered arrays of macromolecules... 

Transmission electron microscopy (TEM) resembles optical microscopy, except that electromagnetic instead of optical lenses are used to focus an electron beam on the sample. Two modes are available in TEM, a bright-freld mode where the intensity of the transmitted beam provides a two-dimensional image of the density or thickness of the sample, and a dark-field mode where the electron diffraction pattern is recorded. A combination of topographic and crystallographic information, including particle size distributions, can be obtained in this way [32],... 

Dark-field electron microscopy (in which the image is formed from the scattered beam), when combined with improved techniques of sample handling and preparation and minimal radiation exposure, can lead to images of sufficiently undamaged DNA at a resolution of 10 A (116). Figure 45 shows such an image in which the two-dimensional projection of the helix is clearly visible on the undamaged part of the molecule. 

High-Angular Annular Dark-Field Scanning Transmission Electron Microscopy... 

O.L. Krivanek, M.F. Chisholm, V. Nicolosi, T.J. Pennycook, G.). Corbin, N. Dellby, et al., Atom-by-atom structural and chemical analysis by annular dark-field electron microscopy, Nature, 464 (2010) 571-574. 

Figure 5-13 Electron micrograph of a DNA molecule (from a bacterial virus bacteriophage T7) undergoing replication. The viral DNA is a long ( 14 pm) duplex rod containing about 40,000 base pairs. In this view of a replicating molecule an internal "eye" in which DNA has been duplicated is present. The DNA synthesis was initiated at a special site (origin) about 17% of the total length from one end of the duplex. The DNA was stained with uranyl acetate and viewed by dark field electron microscopy. Micrograph courtesy J. Wolfson and D. Dressier.

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