Re: [Histonet] mice legs
No, I tend to use my own for work!sorry couldn'r resist midweek madness...;-) On Tue, Sep 15, 2009 at 3:42 PM, Shaw, Sharon shs...@wpi.edu wrote: Good Morning Histo World, I would like to know if anyone is working with mice legs, I have a PI that I work with that wants to process the whole leg, the problem is I need to decal it first and is wondering if the decal will break down the tissue, I think it would he doesn't think so. And if anyone has do this would it be possible to share your protocol with me from decal to processing. Thanks, Sharon- WPI ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet -- Louise Renton Bone Research Unit University of the Witwatersrand Johannesburg South Africa There are nights when the wolves are silent and only the moon howls. George Carlin No trees were killed in the sending of this message. However, many electrons were terribly inconvenienced. ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] mice legs
Good Morning Histo World, I would like to know if anyone is working with mice legs, I have a PI that I work with that wants to process the whole leg, the problem is I need to decal it first and is wondering if the decal will break down the tissue, I think it would he doesn't think so. And if anyone has do this would it be possible to share your protocol with me from decal to processing. Thanks, Sharon- WPI ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
Re: [Histonet] mice legs
Hi Sharon, I frequently get whole mouse legs submitted to me and decalcification doesn't break down the tissue. I use Cal-Rite as a decalcifier but any formic acid decal would be sufficient. Since formic acid is pretty forgiving, I decal the legs for 48-72 hours on a rotator. I change the solution after 24-48 hours and then place them back in decal for 24 hours. I have left bones in decal over the weekend and they have been fine. I have found that if you are focusing on the long bones, this is sufficient but if you are interested in seeing the ankles and feet, you should remove them from the leg and decal them longer. When processing, I use a longer program. Here is the program that I use: 70%1 hr 80%1 hr 95%2 hr 95%2.5 hr 100% 2.5hr 100% 3hr 100% 3hr xylene 1hr20min xylene 1hr20min xylene 1hr20min paraffin 1hr paraffin 1hr paraffin 1.5 hr paraffin 1.5 hr When processing, only process the bone samples with this program. If you batch other tissues in with the bones, the tissue will be over processed. Let me know if you have any more questions. Derek Derek Papalegis HT (ASCP) Senior Histology Technologist Division of Laboratory Animal Medicine Tufts University Shaw, Sharon wrote: Good Morning Histo World, I would like to know if anyone is working with mice legs, I have a PI that I work with that wants to process the whole leg, the problem is I need to decal it first and is wondering if the decal will break down the tissue, I think it would he doesn't think so. And if anyone has do this would it be possible to share your protocol with me from decal to processing. Thanks, Sharon- WPI ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
Re: [Histonet] mice legs
I did a lot of IHC work with whole mouse legs where we had to pay close attention to the tibia, as well as the joint.When the mice were necropsied, I had the prosectors remove all the soft tissue and foot without disrupting the joint. They could do this easily with fresh tissue as opposed to after fixation. The leg bones were fixed for no more than 48 hours, and decalcified in home-brewed 5% formic acid overnight on a shaker table. I then trimmed them to expose the joint and bone marrow prior to processing on a routine program, about 45 minutes per station. Perfect ever single time. IHC was beautiful. Hope this helps. Jackie O' From: Shaw, Sharon shs...@wpi.edu To: histonet@lists.utsouthwestern.edu histonet@lists.utsouthwestern.edu Date: 09/15/2009 08:46 AM Subject: [Histonet] mice legs Sent by: histonet-boun...@lists.utsouthwestern.edu Good Morning Histo World, I would like to know if anyone is working with mice legs, I have a PI that I work with that wants to process the whole leg, the problem is I need to decal it first and is wondering if the decal will break down the tissue, I think it would he doesn't think so. And if anyone has do this would it be possible to share your protocol with me from decal to processing. Thanks, Sharon- WPI ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
