[Histonet] Protocol of MMA Plastic section TRAP staining

Jack Ratliff ratliffjack <@t> hotmail.com
Wed Oct 22 16:48:51 CDT 2014


Hello Dorothy! I will forward you my protocol just as soon as I can get to my computer.

On a side note, I no longer Chair the Hard Tissue Committee for the National Society for Histotechnology (NSH) as I have now been elected to the NSH Board of Directors as the Region III Director for the Southeast region. With that said, Sarah Mack is now the new Hard Tissue Committee Chairperson and Kim Simmons is the Education Development Manager for the NSH. I tell you this because Sarah is located in your part of the country and is currently the Histology Core Manager at University of Rochester Medical Center, Center for Musculosketeal Research. She works with bone and will be an good resource for you. I have copied her on this message. Kim Simmons will be helping to organize bone related educational materials and events for the NSH in the next year so she will be another good reference for you to follow. I have copied her to this message as well. I will still be a part of the Hard Tissue Committee in a supporting role. I just wanted you to know your additional options for information related to the histology of bone, biomaterials and medical device implants.

Best Regards,

Jack Ratliff






> On Oct 22, 2014, at 3:20 PM, Dorothy Hu <abtdhu <@t> gmail.com> wrote:
> 
> Dear Histoneters,
> 
> Can someone email me your TRAP protocol for MMA section? I have one, but
> staining is not consistent. No matter I stain for how long, ~ several hours
> sometimes. There are always variations. I sincerely hope I can get
> different protocol form you and make change. I remember saw other TRAP
> protocol which is much longer than mine.
> 
> The protocol I used was:
> 
> 1. Dissolve 12mg Naphthol AS-MX phosphate (Sigma, N-5000) in 0.5ml N,N
> methylformamide (Sigma,D8654).
> 2. Mix above into 50 ml Sodium tartrate with 0.1M sodium acetate buffer at
> pH5.0.
> 3. Mix 30mg Fast Red Violet LB salt (Sigma, F3381) with above solution and
> filter.
> 
> Stain slides @37oC for at least one hour. The solution can be use for one
> month when stored in 4oC.
> Continue to do counterstain and mount slides.
> Thank you.
> 
> Dorothy Hu
> MGH Endocrine histocore
> Email: dhu5 <@t> partners.org
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