in-situ SEM-observation

[CC]

In article <2mcdpu$1j4 at mserv1.dl.ac.uk>, "Stefan Vetter" 
<Stefan.Vetter at vu-wien.ac.at> wrote:

> Germany). As is generally know, observing organic matter in the SEM requires 
> coating the objekt with a conductive film, mostly gold. Breaking such material, 
> however, automatically exposes uncoated surfaces to the electron beam and leads to 
> strong electric charging even at a low acceleration voltage (1-3 KV) making 
> observation and documentation impossible.
> 
> Has anybody got an idea how we could solve this problem? We would be very grateful 
> for every tip we get!
> 
> Michael Schlag

Hello--
       What you need to do is somehow make all of your material conductive,
not just the surface features.  I can think of two options for this:  
        1)  View the material in a frozen hydrated state, so that the water
will conduct as you are breaking the material.  A problem I can see with
this is that frozen material may not behave the same as unfrozen material. 

        2)  Treat your material in some way to make it conductive.  A few
possibilities are a short fixation followed by postfixation treatment with
osmium tetroxide and thiocarbohydrazide (OTO technique) and/or uranyl
acetate.  You may need to critical point dry or freeze-dry the specimens
afterwards.   We have succesfully viewed uncoated Tetrahymena (lots of
cilia) treated this way at 2.5 kV at upwards of 50kX in our Hitachi 4500.
                                                -- CC