Ad fiber EMs

[PANGELETTI at bmg.bhs.uab.edu]

Hi, since it seemed like a few people were interested in quality EMs of Ad fiber, I went to 
talk to Tom Broker, here at UAB, who has 30 + years of experience in this topic.  I asked 
him about what problems there were in EM of fiber.  

He said: "fiber is very difficult to see.  The main problem is that it is very fragile and 
flexible and tends to break off during sample preparation or imaging." 
He said: "The other problem is that it is very thin and therfore does not stain well with 
uranyl acetate."
He also said that: "Yes, it has been done, but the early work was not of high quality."
I asked him about Cryo-EM (-168oC), and he replied: "It's amazing what an improvement the 
new technology is."  He said that this would definitely help. 
He said: "One improvement might be to use an anti-fiber fAb bound to fiber and try to 
visualize the complex."

So, there you have it.  

		The question that the original poster asked was: what are good techniques to try to 
visualize Ad fiber by EM?  There are many reasons why one would want to do such an 
experiment.  One of which is to improve upon work done earlier.    

**The other point is that observing current methods and technology regarding a problem like 
this maximizes your chances of getting the material published.

I still think that the Stewart, P. et al. (1991) Cell paper is of top notch quality. 
(actually, this paper has the historical references as well)

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