Hi Karl,
        It sounds like you are cutting to cold. Try raising the 
temperature to -10 to -15. The tissues you mention contain a lot of water 
and the ice crystals that form  get harder at the  colder temperatures. 
The softer crystals (warmer) are broken  by your cutting edge when it 
passes through the tissue. If the crystals are to cold, they are scraped 
out of the tissue surface leaving  little craters. Try testing this by 
cutting at 3 to 4 microns. If this "to cold" condition exists. the tissue 
will crumble rather than produce a slice. If you have further questions, 
please call me.
Best regards,
Don 

Don Birgerson
Leica Microsystems
Technical Assistance Center 
Don.Birgerson@Leica-Microsystems.Com
1-800-248-0123  ext 5918 (7:00 to 4:00 CT)



Karl  
Sent by: histonet-bounces@lists.utsouthwestern.edu
11/30/2005 05:07 AM

To
histonet@lists.utsouthwestern.edu
cc

Subject
[Histonet] cryosection bubbles






Dear Histonetters,

When making cryosections of mouse tissues (brain and liver, which is all i
section these days) i get bubble formation between the slide and the
section.

The bubbles are 0.2 -1.0 mm in diameter, and sufficiently large to contact
the blade holder when picking up additional sections on the same slide.

There are no bubbles visible in the OCT, so it seems clear the bubbles are
introduced at the moment the slide is contacting the sections. I get these
bubbles regardless of section thickness (10ums - 25ums).

I'm very satisfied with the sections, but which i could eliminate these
bubbles! Any tips or aspects to pay attention to in this regard are 
greatly
appreciated.

many thanks in advance,

Karl
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