Evaluation of Ferritin Φ X174 and Pyocin as the Morphological Markers for the Immuno-electron Microscopy by the High Resolution Scanning Electron Microscope

Author(s):  
David Joy ◽  
James Pawley

The scanning electron microscope (SEM) builds up an image by sampling contiguous sub-volumes near the surface of the specimen. A fine electron beam selectively excites each sub-volume and then the intensity of some resulting signal is measured. The spatial resolution of images made using such a process is limited by at least three factors. Two of these determine the size of the interaction volume: the size of the electron probe and the extent to which detectable signal is excited from locations remote from the beam impact point. A third limitation emerges from the fact that the probing beam is composed of a finite number of discrete particles and therefore that the accuracy with which any detectable signal can be measured is limited by Poisson statistics applied to this number (or to the number of events actually detected if this is smaller).


2001 ◽  
Vol 7 (S2) ◽  
pp. 822-823
Author(s):  
Stephen K. Chapman

I was trained as a transmission electron microscope engineer in the mid 1960s. I took resolution tests at least once each year and calibrated all of the microscopes that I attended, it was considered a standard procedure for those maintaining an instrument. Moving into the scanning electron microscope field in the mid 1970s it was natural to carry this practice over to that instrument, but in those days this was considered to be extreme. Now, as a consultant in electron microscopy, I routinely carry out SEM resolution, magnification calibration and contamination rate tests on the instruments that I use. I train operators in the role of preventative maintenance and encourage them to know as much as possible about their instruments as this increases their ability to fault find and maintain their own instruments.Resolution - in many laboratories most tungsten hairpin instruments are set up for extended filament life rater than for high resolution.


Author(s):  
G.A.C. Jones ◽  
H. Ahmed ◽  
W.C. Nixon

A special type of post-lens single deflection coils have been developed for use in various applications of micro electron beam technology. The coil geometry has been chosen to give small values of the uncorrectable aberrations and to correct for field curvature by dynamic focusing. With this system 10,000 lines/field may be used in scanning electron microscopy leading to electrical raster shift only to search a specimen. The same type of system could be used to produce high resolution special purpose displays or for direct electron beam photographic recording.In Figure 1 the lettered grid is 3 mm in diameter and the scanned circle is 9 mm in diameter.


Author(s):  
G.G. Hembree ◽  
S.W. Jensen ◽  
J.F. Marchiando

Preliminary results are presented for the measurement of sub-micrometer dimensions of chromium-on-glass lines utilizing a specially modified high resolution scanning electron microscope. The basis of the measurement is an interferometrically positioned, piezoelectrically driven scanning stage. In addition, on-going work in theoretical modeling of the measurement process is di scussed.


Author(s):  
S. Saito ◽  
H. Todokoro ◽  
S. Nomura ◽  
T. Komoda

Field emission scanning electron microscope (FESEM) features extremely high resolution images, and offers many valuable information. But, for a specimen which gives low contrast images, lateral stripes appear in images. These stripes are resulted from signal fluctuations caused by probe current noises. In order to obtain good images without stripes, the fluctuations should be less than 1%, especially for low contrast images. For this purpose, the authors realized a noise compensator, and applied this to the FESEM.Fig. 1 shows an outline of FESEM equipped with a noise compensator. Two apertures are provided gust under the field emission gun.


Author(s):  
J. P. Langmore ◽  
N. R. Cozzarelli ◽  
A. V. Crewe

A system has been developed to allow highly specific derivatization of the thymine bases of DNA with mercurial compounds wich should be visible in the high resolution scanning electron microscope. Three problems must be completely solved before this staining system will be useful for base sequencing by electron microscopy: 1) the staining must be shown to be highly specific for one base, 2) the stained DNA must remain intact in a high vacuum on a thin support film suitable for microscopy, 3) the arrangement of heavy atoms on the DNA must be determined by the elastic scattering of electrons in the microscope without loss or large movement of heavy atoms.


Author(s):  
Mamaeva S.N. ◽  
Vinokurov R.R. ◽  
Munkhalova Ya.A. ◽  
Dyakonova D.P. ◽  
Platonova V.A. ◽  
...  

Currently, due to the intensive development of high-tech science-intensive medical and research devices, more and more attention is paid to the development of diagnostics of rare and difficult to diagnose diseases. It is known that among numerous nephropathies, hematuria may be the only symptom of kidney and urinary tract diseases, which complicates their diagnosis and treatment. In order to develop new approaches for the diagnosis of nephropathies, the authors have been studying the morphology of red blood cells in the blood and urine of children and adults using a scanning electron microscope for several years. The paper presents the results of studies of children with various kidney diseases, including IgA-nephropathy, and chronic glomerulonephritis. Scanning electron microscopy was used for the first time to detect nanoparticles on the surface of red blood cells, the size of which is comparable to the size of viruses, which became the basis for one of the authors ' assumptions, namely, the possible transport of certain types of viruses by red blood cells. Thus, some kidney diseases could be considered virus-associated. This paper presents for the first time the results of determining the glomerular filtration rate of both kidneys separately in the study of separate kidney function and of the study of urine smears obtained during catheterization of the ureters in patients with hydronephrosis of one of the kidneys by scanning electron microscopy. As in previous studies, nanoparticles were found on the surface of red blood cells, which leads to the conclusion about the possible viral nature of the disease of the considered patient. In addition, smear images obtained using a microscope showed a significant difference in the elements of the right and left kidneys urine, which did not contradict the data on the study of glomerular filtration rate. According to the authors, the capabilities of the scanning electron microscope can be applied in fundamental research of kidney diseases at the cellular and molecular levels, forming new ideas about their origin, as well as on the basis of which new methods of non-invasive diagnostics can be built.


Author(s):  
Becky Holdford

Abstract On mechanically polished cross-sections, getting a surface adequate for high-resolution imaging is sometimes beyond the analyst’s ability, due to material smearing, chipping, polishing media chemical attack, etc.. A method has been developed to enable the focused ion beam (FIB) to re-face the section block and achieve a surface that can be imaged at high resolution in the scanning electron microscope (SEM).


Author(s):  
Erik Paul ◽  
Holger Herzog ◽  
Sören Jansen ◽  
Christian Hobert ◽  
Eckhard Langer

Abstract This paper presents an effective device-level failure analysis (FA) method which uses a high-resolution low-kV Scanning Electron Microscope (SEM) in combination with an integrated state-of-the-art nanomanipulator to locate and characterize single defects in failing CMOS devices. The presented case studies utilize several FA-techniques in combination with SEM-based nanoprobing for nanometer node technologies and demonstrate how these methods are used to investigate the root cause of IC device failures. The methodology represents a highly-efficient physical failure analysis flow for 28nm and larger technology nodes.


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.


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