Increasing Access to Tomographic Resources: Web-based Telemicroscopy and Database

2001 ◽  
Vol 7 (S2) ◽  
pp. 92-93
Author(s):  
M. E. Martone ◽  
S. Peltier ◽  
S. Lamont ◽  
A. Gupta ◽  
B. Ludaescher ◽  
...  

The application of electron tomography to cell biology has led to important insights into the 3D fine structure of subcellular processes and organelles. Tomography has been particularly useful for studying relatively large, multi-component structures such as the Golgi apparatus, mitochondria and synaptic complexes. When combined with very powerful high voltage electron microscopes, tomography has also provided high resolution quantitative views of extended structures such as neuronal dendrites in very thick sections (4 μm) at electron microscopic resolution. The utility of tomography is twofold: first, it provides 3D examination of subcellular structure without the need for serial section analysis; second, because the computed slices through the tomographic volumes can be much thinner than is possible to produce by physical sectioning, it reveals structural detail in the range of 5-30 nm that tends to be obscured in conventional thin sections. Tomographic analysis has forced re-assessment of long-standing views of organelles such as mitochondria and the Golgi apparatus and as the technique advances, additional insights are likely forthcoming.Electron tomography is an expensive technique, both in terms of the instruments used and the computational resources required. The three major high voltage electron microscope resources in the United States, San Diego, Boulder and Albany, all are actively engaged in tomographic research and offer this important technology to the scientific community at large.

Author(s):  
A.S. Lossinsky ◽  
M.J. Song

Previous studies have suggested the usefulness of high-voltage electron microscopy (HVEM) for investigating blood-bram barrier (BBB) injury and the mechanism of inflammatory-cell (IC) attachment. These studies indicated that, in evaluating standard conventional thin sections, one might miss cellular attachment sites of ICs in their process of attaching to the luminal endothelial cell (EC) surface of cerebral blood vessels. Our current studies in animals subjected to autoimmune disease suggest that HVEM may be useful in localizing precise receptor sites involved in early IC attachment.Experimental autoimmune encephalomyelitis (EAE) was induced in mice and rats according to standard procedures. Tissue samples from cerebellum, thalamus or spinal cords were embedded in plastic following vascular perfusion with buffered aldehyde. Thick (0.5-0.7 μm) sections were cut on glass knives and collected on Formvar-coated slot grids stained with uranylacetate and lead citrate and examined with the AEI EM7 1.2 MV HVEM in Albany, NY at 1000 kV.


Author(s):  
William H. Massover

Stereoscopic examination of thick sections of fixed and embedded biological tissues by high voltage electron microscopy has been shown to allow direct visualization of three-dimensional fine structure. The present report will consider the occurrence of some new technical problems in specimen preparation and Image interpretation that are not common during lower voltage studies of thin sections.Thick Sectioning and Tissue Coloration - Epon sections of 0.5 μm or more that are cut with glass knives do not have a uniform thickness as Judged by their interference colors; these colors change with time during their flotation on the knife bath, and again when drying onto the specimen support. Quoted thicknesses thus must be considered only as rough estimates unless measured in specific regions by other methods. Chloroform vapors do not always result in good spreading of thick sections; however, they will spread spontaneously to large degrees after resting on the flotation bath for several minutes. Ribbons of thick sections have been almost impossible to obtain.


2003 ◽  
Vol 9 (S03) ◽  
pp. 384-385
Author(s):  
Eileen T. O'Toole ◽  
Kent L. McDonald ◽  
Jana Mäntler ◽  
J. Richard McIntosh ◽  
Anthony A. Hyman ◽  
...  

1997 ◽  
Vol 3 (S2) ◽  
pp. 231-232
Author(s):  
M. E. Martone ◽  
V. M. Edelman ◽  
A. Thor ◽  
S. J. Young ◽  
S. P. Lamont ◽  
...  

Early electron microscopic studies documented that significant changes in the membrane systems of cardiac cells occur in both ischemic and non-ischemic heart failure. These studies relied on analysis of two-dimensional sections and although quantitative changes were observed, the overall organization of the tranverse tubules (T-tubules) and the sarcoplasmic reticulum could not be assessed. In a 3-dimensional study using high voltage electron microscopy (EM) of the T-tubules in spontaneously hypertensive rats, Nakamura and Hama (1991) observed that concomitant with an increase in surface area, the T-tubule system becomes progressively more disorganized and exhibits structural irregularities such as increased numbers of longitudinal tubules, numerous short dead end branches and complex tubular aggregates. These authors suggested that this disorganization may interfere with synchronous contraction over the entire cell.In the present study, we examined the 3-dimensional organization of T-tubules in the left ventricle of explanted human hearts using confocal microscopy and EM tomography.


Sign in / Sign up

Export Citation Format

Share Document