[Histonet] Re: Regarding cartilage and bone staining

[Elizabeth Chlipala]

I'm not sure the size of the mouse head, but first of all the samples needs to be fixed properly I would let it stay in fixative for at least 48 hours.  Then the sample would need to be decaled and that could take days depending upon the type of decal you choose to use.  If you are processing the heads whole your processing cycle (depending upon the size of the samples) could range from 2 to 4 hours per station similar to something like this.  I like to use three absolutes and 3 xylenes on bone samples and find that this works well.  If the samples are over processed you can just cut back on the times.

Liz

70% 2 hours
80% 2 hours
95% 2 hours
100% 2 hours
100% 2 hours
100% 2 hours
Xylene 2 hours
Xylene 2 hours
Xylene 2 hours
Paraffin 2 hours
Paraffin 2 hours
Paraffin 2 hours
Paraffin 2 hours


Elizabeth A. Chlipala, BS, HTL(ASCP)QIHC
Manager
Premier Laboratory, LLC
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Boulder, CO 80308-1592
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-----Original Message-----
From: histonet-bounces <@t> lists.utsouthwestern.edu [mailto:histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of Kraniofacial Biology
Sent: Wednesday, October 12, 2011 1:12 PM
To: histonet <@t> lists.utsouthwestern.edu
Subject: [Histonet] Re: Regarding cartilage and bone staining

Hello
I would like to ask how many time do you put the mice head in xylene after
series of Ethanol wash in the way of making paraffin block. I also did all
the steps as well, but for embryonic day 17-18, even after observing
clearing in xylene and moving to the several times in Gurr/paraplast I find
the head to be soft and the initial part of head from which we start to cut
is white and when I removed the flecks of wax and touched it was still soft.
I am not getting what to do.
Pathik

On Tue, Oct 11, 2011 at 11:01 PM, Kraniofacial Biology <
kraniofacialbiology <@t> gmail.com> wrote:

> Hello Histonetters
> I am new to this site. I am a clinician and very fresh to research. I was
> doing whole mount cartilage and bone staining of whole head of mice and was
> surprised to see whole mouse head being used for experiment. I was wondering
> if there is any other staining method which can be used in place of the
> whole mount cartilage and bone staining that would stain the sections on a
> slide and works on cartilage and bone. This would also save the whole mouse
> to be used and many slides can also be used for other experiments.
> Regards
> Pathik
>
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