Simultaneous Study of Both the Surface and Interior Structure of Biological Specimens in a Scanning Electron Microscope

Author(s):  
P. S. D. Lin ◽  
A. V. Crewe

In contrast to optical and transmission electron microscopes, which probe the interior, the scanning electron microscope usually only reveals the exterior structure of the specimen to the biologist. This is due both to the collection of secondary electrons as signal and to the practice of coating the specimen with various heavy, noble metals.Taking advantage of their penetrating power, one can use backscattered electrons to study the interior structure of the biological specimen in a scanning electron microscope. Here a block of specimen is first stained by means similar to those practiced in transmission electron microscopy. Then a coating of carbon is applied to supress the undesirable specimen charge-up. Although the resolution in this mode will be degraded by electron-atom scattering events, this approach promises to yield more histochemical information than studying merely the topography of the specimen.

1997 ◽  
Vol 3 (S2) ◽  
pp. 1243-1244 ◽  
Author(s):  
Raynald Gauvin ◽  
Steve Yue

The observation of microstructural features smaller than 300 nm is generally performed using Transmission Electron Microscopy (TEM) because conventional Scanning Electron Microscopes (SEM) do not have the resolution to image such small phases. Since the early 1990’s, a new generation of microscopes is now available on the market. These are the Field Emission Gun Scanning Electron Microscope with a virtual secondary electron detector. The field emission gun gives a higher brightness than those obtained using conventional electron filaments allowing enough electrons to be collected to operate the microscope with incident electron energy, E0, below 5 keV with probe diameter smaller than 5 nm. At 1 keV, the electron range is 60 nm in aluminum and 10 nm in iron (computed using the CASINO program). Since the electron beam diameter is smaller than 5 nm at 1 keV, the resolution of these microscopes becomes closer to that of TEM.


Author(s):  
M. Kersker ◽  
C. Nielsen ◽  
H. Otsuji ◽  
T. Miyokawa ◽  
S. Nakagawa

Historically, ultra high spatial resolution electron microscopy has belonged to the transmission electron microscope. Today, however, ultra high resolution scanning electron microscopes are beginning to challenge the transmission microscope for the highest resolution.To accomplish high resolution surface imaging, not only is high resolution required. It is also necessary that the integrity of the specimen be preserved, i.e., that morphological changes to the specimen during observation are prevented. The two major artifacts introduced during observation are contamination and beam damage, both created by the small, high current-density probes necessary for high signal generation in the scanning instrument. The JSM-890 Ultra High Resolution Scanning Microscope provides the highest resolution probe attainable in a dedicated scanning electron microscope and its design also accounts for the problematical artifacts described above.Extensive experience with scanning transmission electron microscopes lead to the design considerations of the ultra high resolution JSM- 890.


Author(s):  
K. Shibatomi ◽  
T. Yamanoto ◽  
H. Koike

In the observation of a thick specimen by means of a transmission electron microscope, the intensity of electrons passing through the objective lens aperture is greatly reduced. So that the image is almost invisible. In addition to this fact, it have been reported that a chromatic aberration causes the deterioration of the image contrast rather than that of the resolution. The scanning electron microscope is, however, capable of electrically amplifying the signal of the decreasing intensity, and also free from a chromatic aberration so that the deterioration of the image contrast due to the aberration can be prevented. The electrical improvement of the image quality can be carried out by using the fascionating features of the SEM, that is, the amplification of a weak in-put signal forming the image and the descriminating action of the heigh level signal of the background. This paper reports some of the experimental results about the thickness dependence of the observability and quality of the image in the case of the transmission SEM.


Author(s):  
S. Takashima ◽  
H. Hashimoto ◽  
S. Kimoto

The resolution of a conventional transmission electron microscope (TEM) deteriorates as the specimen thickness increases, because chromatic aberration of the objective lens is caused by the energy loss of electrons). In the case of a scanning electron microscope (SEM), chromatic aberration does not exist as the restrictive factor for the resolution of the transmitted electron image, for the SEM has no imageforming lens. It is not sure, however, that the equal resolution to the probe diameter can be obtained in the case of a thick specimen. To study the relation between the specimen thickness and the resolution of the trans-mitted electron image obtained by the SEM, the following experiment was carried out.


Author(s):  
Shaopeng Hu ◽  
Jianhua Wang ◽  
Zhen Li ◽  
Huei Chen ◽  
Fei Cu ◽  
...  

Gastritis from returning bile is a common disease, but the reason for the disease is not clear. As the pathologic ultrastructure research progresses, it has drawn attention to the ultrastructural change of cells in gastric mucosa by clinical workers. We observed gastric mucosa tissues of 15 patients suffering from gastritis with a transmission electron microscope (TEM) and a scanning electron microscope (SEM). It is the first report in China that fungus exists in the lamina propria of gastric mucosa tissue. The result is as follows.The gastric mucosa tissues of 15 patients suffering from gastritis were acquired by stomachoscopy. Both TEM and SEM specimens were prepared by the usual methods. Under the TEM, the epithelial surface became higher and larger. Mitochondria of the cells were swollen and cristae were disrupted. There were vacuoles in the cells. The nucleus showed disorder, heterochromatin became darker, and nucleolae could be observed.


Author(s):  
Gao Fengming

Transmission electron microscope(TEM) and scanning electron microscope(SEM) were widely used in experimental tumor studies. They are useful for evaluation of cellular transformation in vitro, classification of histological types of tumors and treating effect of tumors. We have obtained some results as follows:1. Studies on the malignant transformation of mammalian cells in vitro. Syrian golden hamster embryo cells(SGHEC) were transformed in vitro by ThO2 and/or ore dust. In a few days after dust added into medium, some dust crystals were phagocytized. Two weeks later, malignant transformation took place. These cells were of different size, nuclear pleomorphism, numerous ribosomes, increasing of microvilli on cell surface with various length and thickness, and blebs and ruffles(Figs. 1,2). Myelomonocytic leukemic transformation of mouse embryo cells(MEC) was induced in vitro by 3H-TdR. Transformed cells were become round from fusiform. The number of mitochondria and endoplasmic reticulum was reduced, ribosomes and nucleoli increased, shape of nuclei irregular, microvilli increased, and blebs and ruffles appeared(Fig. 3).


Author(s):  
Edward Coyne

Abstract This paper describes the problems encountered and solutions found to the practical objective of developing an imaging technique that would produce a more detailed analysis of IC material structures then a scanning electron microscope. To find a solution to this objective the theoretical idea of converting a standard SEM to produce a STEM image was developed. This solution would enable high magnification, material contrasting, detailed cross sectional analysis of integrated circuits with an ordinary SEM. This would provide a practical and cost effective alternative to Transmission Electron Microscopy (TEM), where the higher TEM accelerating voltages would ultimately yield a more detailed cross sectional image. An additional advantage, developed subsequent to STEM imaging was the use of EDX analysis to perform high-resolution element identification of IC cross sections. High-resolution element identification when used in conjunction with high-resolution STEM images provides an analysis technique that exceeds the capabilities of conventional SEM imaging.


2008 ◽  
Vol 16 (4) ◽  
pp. 62-63
Author(s):  
V.M. Dusevich ◽  
J.H. Purk ◽  
J.D. Eick

Coloring pictures is an educational exercise, which is fun, and helps develop important skills. Coloring SEM micrographs is especially suitable for electron microscopists. Color micrographs are not just great looking on a lab wall; they inspire both microscopists and students to exercise digital picture manipulation. Many microscopists enjoyed looking at the beautiful color micrographs by D. Scharf, but were frustrated to learn they needed a very particular scanning electron microscope equipped with multiple secondary electron detectors in order to color their own pictures. Fortunately, there are other ways to color SEM micrographs. Most SEMs are equipped with at least two detectors, for secondary and backscattered electrons.


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