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Technichal question about frozen sections

Technichal question about frozen sections - Protocols and Methods Forum

Technichal question about frozen sections - Post Any Protocol, Method, Technique, Procedure or Tips / Troubleshooting for any Molecular Biology Technique.


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  #1  
Old 11-17-2005, 01:39 PM
Azy
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Default Technichal question about frozen sections



Hello all

I am a PhD. student at university of ottawa (canada), studying
molecular and cellular medicine. I have a technichal question about
frozen sectioning. In our lab we do immunohistochemistry as a routine
experimental approach and our tissue samples are skin samples. However,

not all the antibodies work on paraffin-embedded sections. Therefore, I

do need to prepare frozen sections of the skin. I have a problem
preparing my frozen blocks which severly reduces the quality of my cut
sections. I use cryomatrix as the embedding medium which is a good
material. To hold the tiny piece of dissected skin as flat and unfolded

as possible, I use a tiny piece of paper towel which functions as a
support for it (attached to the back of the skin), and I embed the
whole thing in the matrix. As the paper towel can not be firmly
attached to the tissue, at the time of sectioning it comes off and
badly damages my tissue. However, if I do not use paper towel, it would

be very difficult to handle the tiny piece of skin which can easily
fold. I know that the skin generally is a more difficult tissue to
handle for frozen sectioning. But, I'm wondering if any of you has
worked with the skin and has any idea about preparing the frozen blocks

or any other approach to help me getting good sections out of it. I
will appreciate your help in advance.
Thanks, Azy

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Old 11-19-2005, 11:23 AM
Sharp Tool
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Default Technichal question about frozen sections

> Hello all
problem

I would just spend the extra time in handling the tissue with care.
If you have alot of tissue to process then I would definately just find
antibodies that work with paraffin wax. However, I don't believe paraffin
wax to be a problem when you can just heat the embedded tissue to melt away
the
wax in and around the tissue allowing the antibodies to bind.

Sharp Tool



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