RE: [Histonet] Undecalcified sample in paraffin and plastic media
Thank you for your information. Since in our lab we have never used MMA and also no vacuum I decided to ask histological service to embed the sample.I will section it and stain them with Von kossa/Alcian blue by myself. The sample is being processed now and i will see if the images of sample embeded in MMA would work better than the paraffin one for my project. The technician told me its hard to obtain a good section embedded in MMA compared to paraffin. Would you give me some tips or protocol for MMA sectioning? Do i need to use adhesive to place the MMA section on the slide? In general does the staining time for paraffin sample work for MMA sample? Thank you, Rui Date: Wed, 2 Oct 2013 11:46:22 -0500 Subject: Re: [Histonet] Undecalcified sample in paraffin and plastic media From: ratliffj...@gmail.com To: ru...@hotmail.com Rui, Did you need any additional assistance? Please let me know if there is anything I can do to be of assistance to you. Best Regards, Jack Jack L Ratliff, Owner/HistologistRatliff Histology Consultants, LLC389 Nichol Mill LaneFranklin, TN 37067 (615) 236-4901 (o)(615) 236-4962 (f)(317) 281-1975 (c) ratliffjack@gmail.comjratl...@ratliffhistology.com jratl...@ratliffhistology.com (coming soon) On Mon, Sep 30, 2013 at 7:49 AM, Jack Ratliff ratliffj...@hotmail.com wrote: Rui, You will definitely want to consider using plastic media like methyl methacrylate (MMA). It will cause less shrinkage in the tissue during polymerization, you can still cut at a range of 4-12 microns using a rotary microtome and tungsten-carbide knife, any mineralization present in the tissue will infiltrate and polymerize well allowing for enhanced stabilization of tissue and section morphology throughout microtomy, and you can even deplastify the sections with certain MMA formulations to increase staining options. Please let me know if you do wish to continue with plastic media as I have helped many labs to get started with and/or to refine their current capabilities with MMA. Additionally, I would like to point out that I Chair the Hard Tissue Committee (HTC) for the National Society for Histotechnology (NSH). Membership with the NSH has several benefits that could also help you to move forward with your project at your own pace. For example, as a member you will have access to all archived publications of the Journal of Histotechnology (JOH). With this access to the JOH via Manny Publishing, the HTC has created a reference document that collates all relevant publications (1970's to present) that pertain to bone, biomaterials, medical device implants, resin histology, etc., so that one can easily locate and obtain publication information relevant to their niche specific needs. Rest assured that I will be happy to help you either way you choose to move forward. Best Regards, Jack On Sep 23, 2013, at 9:19 PM, Rui TAHARA ru...@hotmail.com wrote: I have undecalcified paraffin embed samples that were sectioned at 10 micron that I want to stain with Von kossa. Because samples are embryonic quail heads (ossification starts to happen) and still soft enough to section with standard rotary microtome with tungsten knife in paraffin. My intention is to 3D reconstruct anatomies based on histological sections. Because of this, I am wondering if I should actually use plastic media rather than paraffin to keep the section shape as consistent as possible. Does plastic embed material actually preserve the consistent shape among sections better than paraffin embed sample? No winkle etc..? Is there any other advantage that I actually should use the plastic media than paraffin for what I want to do? I know downside of plastic media is that in general plastic embedding process are lengthy and plastic embedding material are expensive than the paraffin ones, and are mainly use for bone to support the hard material for sectioning. When I sectioned some ossified samples, beak start to fall off from section and the section show the lines from the possibly scratched knife. Is this indication of paraffin media that does not provide enough strength for sectioning? I thought it may possibly the poor infiltration. In our lab nobody has processed the plastic embedding and sectioning (we have only standard microtome, no vaccum machine. Can I section plastic embed sample with the standard microtome at 10 micron?) so I would like to have any input before actually making a plastic embed sample. Any suggestions would be appreciated. Rui TAHARA Biology Department McGill University ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ___ Histonet mailing list Histonet
Re: [Histonet] Undecalcified sample in paraffin and plastic media
Rui, You will definitely want to consider using plastic media like methyl methacrylate (MMA). It will cause less shrinkage in the tissue during polymerization, you can still cut at a range of 4-12 microns using a rotary microtome and tungsten-carbide knife, any mineralization present in the tissue will infiltrate and polymerize well allowing for enhanced stabilization of tissue and section morphology throughout microtomy, and you can even deplastify the sections with certain MMA formulations to increase staining options. Please let me know if you do wish to continue with plastic media as I have helped many labs to get started with and/or to refine their current capabilities with MMA. Additionally, I would like to point out that I Chair the Hard Tissue Committee (HTC) for the National Society for Histotechnology (NSH). Membership with the NSH has several benefits that could also help you to move forward with your project at your own pace. For example, as a member you will have access to all archived publications of the Journal of Histotechnology (JOH). With this access to the JOH via Manny Publishing, the HTC has created a reference document that collates all relevant publications (1970's to present) that pertain to bone, biomaterials, medical device implants, resin histology, etc., so that one can easily locate and obtain publication information relevant to their niche specific needs. Rest assured that I will be happy to help you either way you choose to move forward. Best Regards, Jack On Sep 23, 2013, at 9:19 PM, Rui TAHARA ru...@hotmail.com wrote: I have undecalcified paraffin embed samples that were sectioned at 10 micron that I want to stain with Von kossa. Because samples are embryonic quail heads (ossification starts to happen) and still soft enough to section with standard rotary microtome with tungsten knife in paraffin. My intention is to 3D reconstruct anatomies based on histological sections. Because of this, I am wondering if I should actually use plastic media rather than paraffin to keep the section shape as consistent as possible. Does plastic embed material actually preserve the consistent shape among sections better than paraffin embed sample? No winkle etc..? Is there any other advantage that I actually should use the plastic media than paraffin for what I want to do? I know downside of plastic media is that in general plastic embedding process are lengthy and plastic embedding material are expensive than the paraffin ones, and are mainly use for bone to support the hard material for sectioning. When I sectioned some ossified samples, beak start to fall off from section and the section show the lines from the possibly scratched knife. Is this indication of paraffin media that does not provide enough strength for sectioning? I thought it may possibly the poor infiltration. In our lab nobody has processed the plastic embedding and sectioning (we have only standard microtome, no vaccum machine. Can I section plastic embed sample with the standard microtome at 10 micron?) so I would like to have any input before actually making a plastic embed sample. Any suggestions would be appreciated. Rui TAHARA Biology Department McGill University ___ 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] Undecalcified sample in paraffin and plastic media
I have undecalcified paraffin embed samples that were sectioned at 10 micron that I want to stain with Von kossa. Because samples are embryonic quail heads (ossification starts to happen) and still soft enough to section with standard rotary microtome with tungsten knife in paraffin. My intention is to 3D reconstruct anatomies based on histological sections. Because of this, I am wondering if I should actually use plastic media rather than paraffin to keep the section shape as consistent as possible. Does plastic embed material actually preserve the consistent shape among sections better than paraffin embed sample? No winkle etc..? Is there any other advantage that I actually should use the plastic media than paraffin for what I want to do? I know downside of plastic media is that in general plastic embedding process are lengthy and plastic embedding material are expensive than the paraffin ones, and are mainly use for bone to support the hard material for sectioning. When I sectioned some ossified samples, beak start to fall off from section and the section show the lines from the possibly scratched knife. Is this indication of paraffin media that does not provide enough strength for sectioning? I thought it may possibly the poor infiltration. In our lab nobody has processed the plastic embedding and sectioning (we have only standard microtome, no vaccum machine. Can I section plastic embed sample with the standard microtome at 10 micron?) so I would like to have any input before actually making a plastic embed sample. Any suggestions would be appreciated. Rui TAHARA Biology Department McGill University ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
