Re: [Histonet] Bone Histology Protocol
Trevor, May I ask first if there is a particular reason why you are wanting to decalcify? Have you ever considered resin/plastic embedding of non-decalcified bone? Also, what type/species of bone are you wanting to process? My name is Jack Ratliff and I Chair the Hard Tissue Committee for the National Society for Histotechnology and as a professional histology organization and committee focusing on bone, biomaterials and medical device implants, we offer educational solutions to help those like yourself that are in search of information. This is accomplished through the presentation of workshops at the state, regional and national levels or even by providing free reference materials like processing manuals, books for purchase at non-member or member discounts, and free access to the archives of The Journal of Histotechnology to society members. If any of this interests you please check out the NSH website (www.nsh.org) where you can navigate around to view all of what the society has to offer, become a member and even connect with the Hard Tissue Committee. One last thing I can tell you now is that there will be several bone workshops available at the NSH Symposium/Convention this August in Austin, TX. I will also be one of the speakers at this meeting giving a workshop on resin embedding techniques in support of bone, biomaterials and medical device implants. Best Regards, Jack On Feb 28, 2014, at 10:40 PM, Wait, Trevor Jordan wa...@livemail.uthscsa.edu wrote: Hello colleagues! I'm currently trying to construct a complete Bone Processing Protocol that includes fixation (10%NBF), decalcification (EDTA), Dehydration, Clearing of the decalcificant (Xylene), infiltration, and Embedding in Paraffin. I would like to look at some procedures just to get a good backbone of what a complete procedure is displayed like and I was hoping somebody could give me a website or a source where I could see some. I'm kind of new to bone histological processing so any procedures that are reliable will help! Trevor Jordan Wait University of Texas Health Science Center, San Antonio Class of 2017 MD Candidate Abilene Christian University Class of 2013 Graduate B.S. Biochemistry ___ 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] Bone Histology Protocol
Thanks so much for the reference and I will visit that site and use that site most definitely as a resource. However, the reason I am wanting to decalcify is that my researcher (my employer) has instructed me that that is the way we will be doing it lol. I have read about resin/plastic embedding and that method does seem pretty efficient to use so I'm not sure why he opted to not go the plastic resin route. I do know that we have a limited budget to work with so maybe this is the cheapest route we can afford, along with a microtome that might not be sufficient to cut through hard bone and plastic. As far as the species of bone that we are processing, that too is something I'm not exactly sure about, but is something I should probably find out because from my readings, this is an important aspect. Trevor Jordan Wait University of Texas Health Science Center, San Antonio Class of 2017 MD Candidate Abilene Christian University Class of 2013 Graduate B.S. Biochemistry From: Jack Ratliff ratliffj...@hotmail.com Sent: Saturday, March 1, 2014 3:16 AM To: Wait, Trevor Jordan Cc: histonet@lists.utsouthwestern.edu; Jack Ratliff Subject: Re: [Histonet] Bone Histology Protocol Trevor, May I ask first if there is a particular reason why you are wanting to decalcify? Have you ever considered resin/plastic embedding of non-decalcified bone? Also, what type/species of bone are you wanting to process? My name is Jack Ratliff and I Chair the Hard Tissue Committee for the National Society for Histotechnology and as a professional histology organization and committee focusing on bone, biomaterials and medical device implants, we offer educational solutions to help those like yourself that are in search of information. This is accomplished through the presentation of workshops at the state, regional and national levels or even by providing free reference materials like processing manuals, books for purchase at non-member or member discounts, and free access to the archives of The Journal of Histotechnology to society members. If any of this interests you please check out the NSH website (www.nsh.org) where you can navigate around to view all of what the society has to offer, become a member and even connect with the Hard Tissue Committee. One last thing I can tell you now is that there will be several bone workshops available at the NSH Symposium/Convention this August in Austin, TX. I will also be one of the speakers at this meeting giving a workshop on resin embedding techniques in support of bone, biomaterials and medical device implants. Best Regards, Jack On Feb 28, 2014, at 10:40 PM, Wait, Trevor Jordan wa...@livemail.uthscsa.edu wrote: Hello colleagues! I'm currently trying to construct a complete Bone Processing Protocol that includes fixation (10%NBF), decalcification (EDTA), Dehydration, Clearing of the decalcificant (Xylene), infiltration, and Embedding in Paraffin. I would like to look at some procedures just to get a good backbone of what a complete procedure is displayed like and I was hoping somebody could give me a website or a source where I could see some. I'm kind of new to bone histological processing so any procedures that are reliable will help! Trevor Jordan Wait University of Texas Health Science Center, San Antonio Class of 2017 MD Candidate Abilene Christian University Class of 2013 Graduate B.S. Biochemistry ___ 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] Bone Histology Protocol
I'm terribly sorry, when you asked about species...my inclination was that you were curious which type of bone we were using (femur, humerus, tibia, etc. and cortical/cancellous) but we will be using human bone for this research project that has been donated from various orthopedic surgeries. Trevor Jordan Wait University of Texas Health Science Center, San Antonio Class of 2017 MD Candidate Abilene Christian University Class of 2013 Graduate B.S. Biochemistry From: Jack Ratliff ratliffj...@hotmail.com Sent: Saturday, March 1, 2014 3:16 AM To: Wait, Trevor Jordan Cc: histonet@lists.utsouthwestern.edu; Jack Ratliff Subject: Re: [Histonet] Bone Histology Protocol Trevor, May I ask first if there is a particular reason why you are wanting to decalcify? Have you ever considered resin/plastic embedding of non-decalcified bone? Also, what type/species of bone are you wanting to process? My name is Jack Ratliff and I Chair the Hard Tissue Committee for the National Society for Histotechnology and as a professional histology organization and committee focusing on bone, biomaterials and medical device implants, we offer educational solutions to help those like yourself that are in search of information. This is accomplished through the presentation of workshops at the state, regional and national levels or even by providing free reference materials like processing manuals, books for purchase at non-member or member discounts, and free access to the archives of The Journal of Histotechnology to society members. If any of this interests you please check out the NSH website (www.nsh.org) where you can navigate around to view all of what the society has to offer, become a member and even connect with the Hard Tissue Committee. One last thing I can tell you now is that there will be several bone workshops available at the NSH Symposium/Convention this August in Austin, TX. I will also be one of the speakers at this meeting giving a workshop on resin embedding techniques in support of bone, biomaterials and medical device implants. Best Regards, Jack On Feb 28, 2014, at 10:40 PM, Wait, Trevor Jordan wa...@livemail.uthscsa.edu wrote: Hello colleagues! I'm currently trying to construct a complete Bone Processing Protocol that includes fixation (10%NBF), decalcification (EDTA), Dehydration, Clearing of the decalcificant (Xylene), infiltration, and Embedding in Paraffin. I would like to look at some procedures just to get a good backbone of what a complete procedure is displayed like and I was hoping somebody could give me a website or a source where I could see some. I'm kind of new to bone histological processing so any procedures that are reliable will help! Trevor Jordan Wait University of Texas Health Science Center, San Antonio Class of 2017 MD Candidate Abilene Christian University Class of 2013 Graduate B.S. Biochemistry ___ 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
[Histonet] RE: Bone histotechniques
Thank you so much for the materials, they were definitely helpful for the EDTA and I very much appreciate that! Yes I have contacted my Researcher, unfortunately it is the weekend whenever I started to research for this protocol so my answers may not come til Monday morning. However, I do know that our species is human bones that have been donated from various orthopedic surgeries. As far as the exact type of bone (tibia, femur, etc.) I do not know the answer for they are just shavings or scraps that have been conserved from the surgeries that would otherwise be exposed of. I will also obtain that information as well. Trevor Jordan Wait University of Texas Health Science Center, San Antonio Class of 2017 MD Candidate Abilene Christian University Class of 2013 Graduate B.S. Biochemistry From: gayle callis gayle.cal...@bresnan.net Sent: Saturday, March 1, 2014 10:13 AM To: Wait, Trevor Jordan Subject: Bone histotechniques Trevor, Please say what kind of bone you are working with, including species, femur, tibia, etc. Attached is an excellent EDTA decalcification that was developed years ago by a bone expert/professor, and is published. Once you give more bone details, then I can expound on how to do this. You will probably get many replies. Also, are you using automated tissue processing or have to do this by hand? What are you to do for staining? Immunohistochemistry? Other staining only? There are a lot of variables here that affect how you fix, etc for the bone you are working with. Gayle Callis HTL/HT/MT(ASCP) ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet