Studying membrane proteins in intact cells using nanoparticle labels and liquid-phase electron microscopy

Author(s):  
Niels de Jonge ◽  
Diana Peckys
Author(s):  
C.D. Humphrey ◽  
T.L. Cromeans ◽  
E.H. Cook ◽  
D.W. Bradley

There is a variety of methods available for the rapid detection and identification of viruses by electron microscopy as described in several reviews. The predominant techniques are classified as direct electron microscopy (DEM), immune electron microscopy (IEM), liquid phase immune electron microscopy (LPIEM) and solid phase immune electron microscopy (SPIEM). Each technique has inherent strengths and weaknesses. However, in recent years, the most progress for identifying viruses has been realized by the utilization of SPIEM.


Author(s):  
F. Banhart ◽  
F.O. Phillipp ◽  
R. Bergmann ◽  
E. Czech ◽  
M. Konuma ◽  
...  

Defect-free silicon layers grown on insulators (SOI) are an essential component for future three-dimensional integration of semiconductor devices. Liquid phase epitaxy (LPE) has proved to be a powerful technique to grow high quality SOI structures for devices and for basic physical research. Electron microscopy is indispensable for the development of the growth technique and reveals many interesting structural properties of these materials. Transmission and scanning electron microscopy can be applied to study growth mechanisms, structural defects, and the morphology of Si and SOI layers grown from metallic solutions of various compositions.The treatment of the Si substrates prior to the epitaxial growth described here is wet chemical etching and plasma etching with NF3 ions. At a sample temperature of 20°C the ion etched surface appeared rough (Fig. 1). Plasma etching at a sample temperature of −125°C, however, yields smooth and clean Si surfaces, and, in addition, high anisotropy (small side etching) and selectivity (low etch rate of SiO2) as shown in Fig. 2.


1996 ◽  
Vol 109 (6) ◽  
pp. 1215-1227 ◽  
Author(s):  
I. Hemery ◽  
A.M. Durand-Schneider ◽  
G. Feldmann ◽  
J.P. Vaerman ◽  
M. Maurice

In hepatocytes, newly synthesized apical plasma membrane proteins are first delivered to the basolateral surface and are supposed to reach the apical surface by transcytosis. The transcytotic pathway of apical membrane proteins and its relationship with other endosomal pathways has not been demonstrated morphologically. We compared the intracellular route of an apical plasma membrane protein, B10, with that of polymeric IgA (pIgA), which is transcytosed, transferrin (Tf) which is recycled, and asialoorosomucoid (ASOR) which is delivered to lysosomes. Ligands and anti-B10 monoclonal IgG were linked to fluorochromes or with peroxidase. The fate of each ligand was followed by confocal and electron microscopy in polarized primary monolayers of rat hepatocytes. When fluorescent anti-B10 IgG and fluorescent pIgA were simultaneously endocytosed for 15–30 minutes, they both uniformly labelled a juxtanuclear compartment. By 30–60 minutes, they reached the bile canaliculi. Tf and ASOR were also routed to the juxtanuclear area, but their fluorescence patterns were more punctate. Microtubule disruption prevented all ligands from reaching the juxtanuclear area. This area corresponded, at least partially, to the localization of the mannose 6-phosphate receptor, an endosomal marker. By electron microscopy, the juxtanuclear compartment was made up of anastomosing tubules connected to vacuoles, and was organized around the centrioles. B10 and pIgA were mainly found in the tubules, whereas ASOR was segregated inside the vacuolar elements and Tf within thinner, recycling tubules. In conclusion, transcytosis of the apical membrane protein B10 occurs inside tubules similar to those carrying pIgA, and involves passage via the pericentriolar area. In the pericentriolar area, the transcytotic tubules appear to maintain connections with other endosomal elements where sorting between recycled and degraded ligands occurs.


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