[Histonet] Re:Frozen Spleen w/bubbles

From:Gayle Callis

Jo Dee,

I have questions first.

What species is the spleen from?

How thick are the sections?

What temperature are you sectioning at?  i.e. knife 
temperature?  Block/tissue temperature?

How to you pick up a FS onto the slide?  Slide to section so it comes onto 
glass surface, another method?

What kind of blades are you using?

Are you sections perfectly flat under antiroll device or by brush method, 
whichever you use?

What kind of slides are you using?  Plus Charge?

Before IHC do you air dry your sections at RT or overnight before fixation 
with what????

How are you fixing your spleens?

Do you use a peroxidase blocker that has a high concentration of hydrogen 

At 03:30 PM 3/30/2005, you wrote:
>I am cryosectioning unfixed fresh frozen spleens and when they are 
>sectioned I can see a billion bubbles under the section.  Can anyone explain-
>1.  What is happening?
>2.  How can I avoid them?
>3.  Is there a fixation procedure that can help?
>The sections are falling off of the slide during staining.
>Thank you for your help,
>Jo Dee
>Jo Dee Fish
>Research Technologist III
>Gladstone Institute of Cardiovascular Disease

Gayle Callis
Research Histopathology Supervisor
Veterinary Molecular Biology
Montana State University - Bozeman
PO Box 173610
Bozeman MT 59717-3610
406 994-6367 (lab with voice mail)
406 994-4303 (FAX)

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