High-Resolution Scanning Electron Microscopy for the Imaging of Nuclear Pore Complexes and Ran-Mediated Transport

Author(s):  
Lihi Shaulov ◽  
Boris Fichtman ◽  
Amnon Harel
1977 ◽  
Vol 72 (1) ◽  
pp. 118-132 ◽  
Author(s):  
R H Kirschner ◽  
M Rusli ◽  
T E Martin

We have used high resolution scanning electron microscopy (SEM) to study the nuclear envelope components of isolated mouse liver nuclei. The surfaces of intact nuclei are covered by closely packed ribosomes which are distinguishable by SEM from nuclear pore complexes. After removal of nuclear membranes with the nonionic detergent Triton X-100, the pore complexes remain attached to an underlying, peripheral nuclear lamina, as described by others. The surface of this dense lamina is composed of particulate granules, 75-150 A in diameter, which are contiguous over the entire periphery. We did not observe the pore-to-pore fibril network suggested by other investigators, but such a structure might be the framework upon which the dense lamina is formed. Morphometric analysis of pores and pore complexes shows their size, structure, and density to be similar to that of other mammalian cells. In addition, several types of pore complex-associated structures, not previously reported by other electron microscope (EM) techniques, are observed by SEM. Our studies suggest that the major role of the dense lamina is associated with the distribution, stability, and perhaps, biogenesis of nuclear pore complexes. Treatment of isolated nuclei with a combination of Triton X-100 and sodium deoxycholate removes membranes, dense lamina, and nuclear pore complexes. The resulting "chromatin nuclei" retain their integrity despite the absence of any limiting peripheral structures.


1992 ◽  
Vol 119 (6) ◽  
pp. 1429-1440 ◽  
Author(s):  
M W Goldberg ◽  
T D Allen

The nuclear envelope (NE) of amphibian oocytes can be readily isolated in relatively structurally intact and pure form and has been used extensively for structural studies. Using high resolution scanning electron microscopy (HRSEM), both surfaces of the NE can be visualized in detail. Here, we demonstrate the use of HRSEM to obtain high resolution information of NE structure, confirming previous data and providing some new information. NEs, manually isolated from Triturus cristatus oocytes, have been mounted on conductive silicon chips, fixed, critical point dried and coated with a thin, continuous film of chromium or tantalum and viewed at relatively high accelerating voltage in a field emission scanning electron microscope with the sample within the objective lens. Both nucleoplasmic and cytoplasmic surfaces of the nuclear pore complexes (NPC) have been visualized, revealing the cytoplasmic coaxial ring, associated particles, central plug/transporter and spokes. The nucleoplasmic face is dominated by the previously described basketlike structure attached to the nucleoplasmic coaxial ring. In Triturus, a novel, highly regular flat sheet of fibers, termed the NE lattice (NEL) has been observed attached to the distal ring of the NPC basket. The NEL appears to be distinct from the nuclear lamina. Evidence for the NEL is also presented in thin TEM sections from Triturus oocytes and GVs and in spread NEs from Xenopus. A model is presented for NEL structure and its interaction with the NPCs is discussed.


Scanning ◽  
1997 ◽  
Vol 19 (6) ◽  
pp. 403-410 ◽  
Author(s):  
T. D. Allen ◽  
G. R. Bennion ◽  
S. A. Rutherford ◽  
S. Reipert ◽  
A. Ramalho ◽  
...  

Author(s):  
D. Johnson ◽  
P. Moriearty

Since several species of Schistosoma, or blood fluke, parasitize man, these trematodes have been subjected to extensive study. Light microscopy and conventional electron microscopy have yielded much information about the morphology of the various stages; however, scanning electron microscopy has been little utilized for this purpose. As the figures demonstrate, scanning microscopy is particularly helpful in studying at high resolution characteristics of surface structure, which are important in determining host-parasite relationships.


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).


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