[Histonet] Problems in processing rat paws

Catia Lopes catialopes43 <@t> gmail.com
Fri Nov 18 11:04:57 CST 2011

Hello everyone!
I have a problem in processing rat paws to paraffin embedding.
I need to process and cut the whole paw (except fingers). So, first I need
to decalcify the paw and then process it to paraffin embedding.
My problem is that when I try to cut the paw I can not get a good section
because the tissue have a gum-like consistence (mushy).
Do you know where is my problem? Dehydration, clearing, embedding?
*Note*: I decalcify the paws in  formic acid and aluminum chloride for 24h;
Then I wash samples in phosphate buffer saline for 5 hours. After that I
did the following manual processing (applying vacuum): 2x30min in 70%
ethanol, 2x30min in 80% ethanol, 2x30min in 96% ethanol, 2x30min in
absolute ethanol, 3x15 min in Xilene, 2x15min in paraffin and a final
incubation of 30min in paraffin.
Then, I proceed to paraffin ambedding and try to cut sections of 5um of

If anyone can help me I will be grateful.
Thanks in advance,

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