immunogold labeling, EM questions

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     It's been a while since I have done any EM so I am not really up to date 
with techniques, so I have a few questions for anyone interested in 
 1.  In paraffin,  I believe the avidin/streptavidin-biotin method is 
considered more sensitive than the direct  method (labeled antibody binds to 
antigen). Is everyone using this ABC technique now for immunogold work as seems like it should be better, but does anyone have first hand 

   2.  In sectioning lowicryl, do you guys have any special methods for 
dealing with it. I have never sectioned it.  For .5 um sections do you still 
heat the section on a hot plate to spread the section like you would do with 

   3.  If I am not interested in low temperature, what would be the reason to 
choose  LR White over  Lowicryl or vise versa for immuno?

   4.  More specifically, if anyone out there is using immunogold to identify 
collagen, do you recommend using the smaller gold size of 5 or 1 nm vs the 10 

Thanks for your input. I am glad I am getting the chance to do some EM again 
:) ,


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