[Histonet] help ! paraffin section
Rene J Buesa
rjbuesa <@t> yahoo.com
Wed Aug 8 09:48:32 CDT 2012
What you describe is a typical example of poor paraffin infiltration = the paraffin has not infiltrated the tissue and when you prepare the final block it will consist of 2 different components; the tissue and the paraffin. That is why you end with a "good paraffin section" without the tissue.
Poor paraffin infiltration is always caused by an improper sequence while tissue processing. Either the fixation is incomplete
OR the dehydration is incomplete and there is water in the tissue when you go to the "clearing" stage
OR the clearing stage is incomplete and the tissue still has alcohol (immiscible with paraffin) when the tissue goes to the paraffin
OR the paraffin infiltration is too short.
The problem resides in your processing protocol and there is nothing you can do about that at the end.
Try to check your processing protocol to eliminate the problem.
If this is happening "all of the sudden" while you used to have good results previously, then you either have changed reagents or the reagents are not in a good condition.
From: Megha Kumar <meghak <@t> g.clemson.edu>
To: histonet <@t> lists.utsouthwestern.edu
Sent: Tuesday, August 7, 2012 11:45 PM
Subject: [Histonet] help ! paraffin section
I am trying to section adult mouse intestine and skin using paraffin
embedding. However, when i section, the tissue is torn although the rest of
the paraffin looks perfect. Please suggest why this is happening. Also,
sometimes the skin sections fall off the slides when I perform in situ
hybridization. Any ideas how to prevent this?
Please help! i am a beginner in histology and dont' know what to do!
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