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Scanning electron microscopy is providing increasingly definitive solutions to criminal problems since its commercial debut in 1965. This has been due to the ability of the scanning electron microscope (SEM) to simultaneously produce several electron probe‐induced signals from the specimen, which generate readily interpretable images of surface topography and material composition. The successful applications of these signals are determined by sample preparation and instrumental parameters. No other microbeam technology combines high resolution (2–5 nm) of the topographic (secondary) electrons with large depth of field for three‐dimensional viewing. The SEM is indeed ideal for stereomicroscopy. The versatility of the SEM stems from its additional capability to process each specimen signal by various contrast enhancement methods, such as line scanning, deflection modulation (DM), area mapping, etc. These methods allow an intuitive, stylistic, and synthetic analysis of the image and are ideal for quality control analysis. Digital SEMs have pioneered in automated image processing and unattended search and analysis of particulates. The combined SEM and energy‐dispersive X‐ray microanalysis (EDX) is the most definitive technique in testing for gunshot residue (GSR) particles, collected by the glue‐lift technique. In the analyses of other trace evidence, such as hair and fibers, in physical matching, and in nondestructive elemental analysis of physical evidence, the SEM/EDX is the most efficient of all microbeam technologies. From firearms, bullet wounds and human bones, to plants, pollen and fungi, the list of criminal evidence examined by SEM/EDX is endless. However, the SEM/EDX is not ideal for quantitative analysis of elements present as traces (<1% w/w). The SEM lacks the three‐dimensional sectioning abilities of scanning beam confocal microscopes. The imaging capabilities of the SEM surpass these limitations. The SEM is, therefore, a major tool in forensic research and investigation.
Scanning electron microscopy is providing increasingly definitive solutions to criminal problems since its commercial debut in 1965. This has been due to the ability of the scanning electron microscope (SEM) to simultaneously produce several electron probe‐induced signals from the specimen, which generate readily interpretable images of surface topography and material composition. The successful applications of these signals are determined by sample preparation and instrumental parameters. No other microbeam technology combines high resolution (2–5 nm) of the topographic (secondary) electrons with large depth of field for three‐dimensional viewing. The SEM is indeed ideal for stereomicroscopy. The versatility of the SEM stems from its additional capability to process each specimen signal by various contrast enhancement methods, such as line scanning, deflection modulation (DM), area mapping, etc. These methods allow an intuitive, stylistic, and synthetic analysis of the image and are ideal for quality control analysis. Digital SEMs have pioneered in automated image processing and unattended search and analysis of particulates. The combined SEM and energy‐dispersive X‐ray microanalysis (EDX) is the most definitive technique in testing for gunshot residue (GSR) particles, collected by the glue‐lift technique. In the analyses of other trace evidence, such as hair and fibers, in physical matching, and in nondestructive elemental analysis of physical evidence, the SEM/EDX is the most efficient of all microbeam technologies. From firearms, bullet wounds and human bones, to plants, pollen and fungi, the list of criminal evidence examined by SEM/EDX is endless. However, the SEM/EDX is not ideal for quantitative analysis of elements present as traces (<1% w/w). The SEM lacks the three‐dimensional sectioning abilities of scanning beam confocal microscopes. The imaging capabilities of the SEM surpass these limitations. The SEM is, therefore, a major tool in forensic research and investigation.
Cathodoluminescence (CL) is the light that is emitted from a material irradiated by an electron beam. The present study was undertaken to show the applicability to biological studies of a scanning electron microscope (SEM) equipped with a high-sensitive cathodoluminescence detection system. For this purpose, we injected inorganic fluorescent powders (P43) suspended in phosphate buffered saline into rat blood circulation, fixed the animals with glutaraldehyde within a day, and observed the hepatic tissues with a SEM. Our instrument enabled the simultaneous collection of both secondary electron (SE) and CL images of these tissues. Backscattered electron (BSE) images of the same portion were also able to be obtained with this microscope. SE and BSE images clearly showed the three-dimensional structure of the hepatic tissues including hepatocytes, Kupffer cells, Ito cells, and sinusoidal epithelial cells, while CL images visualized cathodoluminescence signals emitted from P43 as bright spots. We observed non-coated tissues under a low-vacuum condition and metal-coated tissues under a high-vacuum condition, and found that the high-vacuum observation of metal-coated tissues provided high quality CL images of P43 in the Kupffer cells. The superimposition of the CL images onto the corresponding SE or BSE images revealed that bright spots in the CL images were produced by the fluorescent powders uptaken by Kupffer cells. These findings indicate that the detection of CL as well as SE or BSE signals by SEM all provide us with useful information on the distribution of fluorescent tracers in tissues and cells in three-dimensional images.
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