A polymer designed to optimize antigen localization in immunoelectron microscopy

Author(s):  
J.B. Olesen ◽  
C.A. Heckman ◽  
A. Lukinius

We have developed a new polymer for the purpose of retaining the maximal antigenicity of proteins in samples embedded for electron microscopy. HACH is a mixture of hydroxyhexanedial, carbohydrazide and hydrazine that polymerizes at room temperature within 24 h. Tissues embedded in the polymer were thin sectioned and the sections, mounted on formvar films, were viewed with a Zeiss IOC transmission electron microscope. The ultrastructural features of cells in HACH-embedded tissues appeared similar to those previously found in GACH-embedded tissues.The ability to preserve the antigenicity of a protein up to the polymerization stage has been tested in the following manner. Human red blood cells, from either Rh+ or Rh- donors, were first exposed to the HACH prepolymer and then rinsed and exposed to a commercially available anti-D antibody. After removal of the prepolymer, Rh+ red blood cells were clumped by anti-D antibody to the same extent as control, untreated cells (Fig. 1).

Author(s):  
A.J. Tousimis ◽  
T.R. Padden

The size, shape and surface morphology of human erythrocytes (RBC) were examined by scanning electron microscopy (SEM), of the fixed material directly and by transmission electron microscopy (TEM) of surface replicas to compare the relative merits of these two observational procedures for this type specimen.A sample of human blood was fixed in glutaraldehyde and washed in distilled water by centrifugation. The washed RBC's were spread on freshly cleaved mica and on aluminum coated microscope slides and then air dried at room temperature. The SEM specimens were rotary coated with 150Å of 60:40- gold:palladium alloy in a vacuum evaporator using a new combination spinning and tilting device. The TEM specimens were preshadowed with platinum and then rotary coated with carbon in the same device. After stripping the RBC-Pt-C composite film, the RBC's were dissolved in 2.5N HNO3 followed by 0.2N NaOH leaving the preshadowed surface replicas showing positive topography.


Author(s):  
Robert W. Weise

The role that scanning electron microscopy (SEM) is playing in descriptive helminthology is becoming more apparent in the literature. However, the majority of papers on the SEM of helminths have used conventional or modified light microscope techniques of fixation and dehydration, and not established SEM techniques in which freeze- and critical point-drying are routinely used. The present investigation was undertaken to examine the applicability of modified scanning and transmission electron microscope techniques for the preparation of certain helminths for SEM.Method I.– Live animal-parasitic nematodes were fixed in 6% phosphate buffered glutaraldehyde for 24 hr at room temperature.


Author(s):  
J. Doerschel

AbstractDislocation configurations induced by room temperature microindentations on the (001) face of GaSb (undoped and Te-doped) have been studied using high voltage transmission electron microscopy. Perfect and partial dislocations could be found in all four arms of the dislocation rosette around the indent. Microtwins and rarely single stacking faults are associated with the partials. Contrary to other binary III–V compounds, an “inverse” glide prism along the [1[unk]0]/[[unk]10] rosette arms is created and it is bounded by {111}


Author(s):  
M.A. O’Keefe ◽  
J. Taylor ◽  
D. Owen ◽  
B. Crowley ◽  
K.H. Westmacott ◽  
...  

Remote on-line electron microscopy is rapidly becoming more available as improvements continue to be developed in the software and hardware of interfaces and networks. Scanning electron microscopes have been driven remotely across both wide and local area networks. Initial implementations with transmission electron microscopes have targeted unique facilities like an advanced analytical electron microscope, a biological 3-D IVEM and a HVEM capable of in situ materials science applications. As implementations of on-line transmission electron microscopy become more widespread, it is essential that suitable standards be developed and followed. Two such standards have been proposed for a high-level protocol language for on-line access, and we have proposed a rational graphical user interface. The user interface we present here is based on experience gained with a full-function materials science application providing users of the National Center for Electron Microscopy with remote on-line access to a 1.5MeV Kratos EM-1500 in situ high-voltage transmission electron microscope via existing wide area networks. We have developed and implemented, and are continuing to refine, a set of tools, protocols, and interfaces to run the Kratos EM-1500 on-line for collaborative research. Computer tools for capturing and manipulating real-time video signals are integrated into a standardized user interface that may be used for remote access to any transmission electron microscope equipped with a suitable control computer.


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.


Alloys of Al-5% Pb and Al-5% Pb-0.5% Si (by mass) have been manufactured by rapid solidification and then examined by transmission electron microscopy. The rapidly solidified alloy microstructures consist of 5-60 nm Pb particles embedded in an Al matrix. The Pb particles have a cube-cube orientation relation with the Al matrix, and are cub-octahedral in shape, bounded by {100} Al, Pb and {111} Al, Pb facets. The equilibrium Pb particle shape and therefore the anisotropy of solid Al-solid Pb and solid Al-liquid Pb surface energies have been monitored by in situ heating in the transmission electron microscope over the temperature range between room temperature and 550°C. The ani­sotropy of solid Al-solid Pb surface energy is constant between room temperature and the Pb melting point, with a {100} Al, Pb surface energy about 14% greater than the {111} Al, Pb surface energy, in good agreement with geometric near-neighbour bond energy calculations. The {100} AI, Pb facet disappears when the Pb particles melt, and the anisotropy of solid Al-liquid Pb surface energy decreases gradually with increasing temperature above the Pb melting point, until the Pb particles become spherical at about 550°C.


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