Help:electron microscopy of adenovirus

PANGELETTI at bmg.bhs.uab.edu PANGELETTI at bmg.bhs.uab.edu
Thu Sep 7 12:50:17 EST 1995



Hi Sophia, what you have encountered is a problem that other Adenovirus researchers such as 
T. Broker and L. Chow have seen.  The problem is that fiber is very flexible.  Ad 2 and 5 
have fairly short fiber extentions Ad 31 fiber is quite a bit longer - and you can imagine 
what this means.  Essentially, the problem is that the electron beam causes the fiber 
proteins to oscillate. So actually its a case of Eisenberg's uncertainty principle - by 
attempting to observe the protein it becomes unobservable.  So, it's rather difficult.
I think the best paper I have seen regarding Adenovirus capsid structure was:
Stewart et al. Cell 67:145-154.  I am not a microscopist, but I noticed that they used 
-168oC for capturing images.  Maybe low T would help. 

Anyway, good luck. 

PCA


I am trying to get clear micrographs of the fine fiber proteins which extend from 
adenoviruses.
I am currently using carbon-coated grids and a Philips300 EM and have found that staining 
the virus with sodium silicotungstate produces very nice definition of the virus capsid, 
however, the fibres are seldom visible.  Also, the few times I do see the fibres, they don't 
show up on the micrographs, or when they do they are very blurry.  

 I  have noticed that when taking pictures, the carbon film tends to drift.  If you have any 
suggestions on how to prevent this drift or know of any other techniques for adenovirus 
fiber staining, I would be very grateful. 

Sophia Mittas
sophia.mittas at utoronto.ca




Peter C. Angeletti
Department of Biochemistry and Molecular Genetics
University of Alabama at Birmingham 
)>>-<(()):)
Pangeletti at bmg.bhs.uab.edu








More information about the Virology mailing list