Name: undecalcified bone preparation for histology, histomorphometry and fluorochrome analysis
Uploaded: 2010-01-08T17:00:00
Description: Watch this Scientific Journal Video about Undecalcified Bone Preparation for Histology, Histomorphometry and Fluorochrome Analysis at JoVE.com

<p class="jove_content">作者要感谢她的专长苏康奈尔女士的协助下，在树脂嵌入和她的实验室技术女士千金墓群。作者要感谢他们的意见弗兰克Kandziora教授和博士玛丽 - 安妮Polboth。 </p>

<ol><li>该组织是放置在一个密封的容器中含有10％磷酸<sup>的</sup>福尔马林溶液3。确保尽量减少暴露在光线下，如果要执行，作为荧光光敏感荧光分析。 </li><li>建立将需要组织的方向。 </li><li>修剪大小的标本，在正确的方向，用一个带锯。确保安全措施得到遵守。试样，然后将放在不透明的容器，其中的量应该是10倍左右大小的标本，以达到充分固定。试样应保持在福尔马林溶液中的一到两个星期取决于其大小总?...

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	<li>Kerr, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> <em style='font-style: italic'>Atlas of Functional Histology</em>. , 164-168 (2000).</li><li>Parfitt, A. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Bone+histomorphometry:+standardization+of+nomenclature,+symbols,+and+units.+Report+of+the+ASBMR+Histomorphometry+Nomenclature+Committee.">Bone histomorphometry: standardization of nomenclature, symbols, and units. Report of the ASBMR Histomorphometry Nomenclature Committee.</a> <em style='font-style: italic'>J Bone Miner Res</em>. <b>2</b>, 595-610 (1987).</li><li>An, Y. H., Martin, K. l. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> <em style='font-style: italic'>Handbook of histology methods for bone and cartilage</em>. , (2003).</li><li>Bancroft, J. D., Gamble, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> <em style='font-style: italic'>Theory and Practice of Histological Techniques</em>. , 352-360 (2008).</li></ol>

<u>溶剂：</u> <ul><li>酸，10％的正常缓冲福尔马林，乙醇，二甲苯和丁醇<br />所有组织学级，无硫。 <br /> Grale科学Pty有限公司<br /> http://www.grale.com.au </li></ul> <u>模具：</u> <ul><li>剥离，塑料模具<br /> Grale科学</li></ul> <u>幻灯片：</u> <ul><li> SUPERFROST PLUS的显微镜幻灯片<br />博士伦科学Pty有限公司<br /> <a href="http://www.lomb.com.au/">http://www.lomb.com.au</a> </li></ul> <u>树脂：</u> <ul><li> GlycolMethacrylate Technovit 7100 <br />贺利氏古莎<br /> <a href="http://www.kulzer-technik.de/168/Products/Histology/Technovit_7100.htm">http://www.kulzer-technik.de/168/Products/Histology/Technovit_7100.htm</a> </li><li>背座树脂Technovit 3040 HISTO块<br />贺利氏古莎</li></ul> <u>渍</u> <br /> <span>（Grale科学试剂）</span> <p> Goldner三色染色</li><p style="margin-left:40px;"> 1。 Weigert氏铁hemotoxylin（DeGroat） </p><table style="margin-left:40px;" border="0"><tr><td colspan="2" style="font-weight:bold;"> A）Hemotoxylin解决方案</td></tr><tr><td> Hemotoxylin </td><td> 1克</td></tr><tr><td>无水酒精</td><td> 100毫升</td></tr><tr><td colspan="2"  style="font-weight:bold;"> B）铁溶液</td></tr><tr><td> 30％三氯化铁水溶液</td><td> 4ML </td></tr><tr><td>盐酸（浓） </td><td> 1ML </td></tr><tr><td>蒸馏水</td><td> 95ml置于</td></tr></table><p style="margin-left:40px;" >过滤和添加到同等体积的hemotoxylin立即前染色使用。 </p><p style="margin-left:40px;" > 2。丽春红，品红azophloxin股票解决方案</p><table style="margin-left:40px;" border="0"><tr><td colspan="2"  style="font-weight:bold;"> A）丽春红 - 二甲基苯胺的解决方案</td></tr><tr><td>丽春红 - 二甲基苯胺</td><td> 0.75克</td></tr><tr><td>酸fuchsion </td><td> 0.25克</td></tr><tr><td>醋酸</td><td> 1ML </td></tr><tr><td colspan="2">混合，并添加蒸馏水（100毫升） </td></tr><tr><td colspan="2"  style="font-weight:bold;"> B）azophloxin解决方案</td></tr><tr><td> Azophloxin </td><td> 0.5克</td></tr><tr><td>乙酸</td><td> 0.6mL </td></tr><tr><td colspan="2">混合，并添加蒸馏水（100毫升） </td></tr><tr><td colspan="2"  style="font-weight:bold;"> c）最后的工作染色解决方案</td></tr><tr><td>丽春红，品红溶液</td><td> 5 - 10毫升</td></tr><tr><td> Azophloxin </td><td> 2ML </td></tr><tr><td> 0.2％的醋酸溶液</td><td> 88mL </td></tr></table><p style="margin-left:40px;"> 3。浅绿色解决方案</p><table style="margin-left:40px;" border="0"><tr><td>浅绿色</td><td> 1克</td></tr><tr><td>乙酸</td><td> 1ML </td></tr><tr><td colspan="2">混合，并添加蒸馏水（500毫升） </td></tr></table><p style="margin-left:40px;"> 4。 Phophomolybdic酸性橙G解决方案</p><table style="margin-left:40px;" border="0"><tr><td>磷钼酸</td><td> 3克</td></tr><tr><td>橙G </td><td> 2克</td></tr><tr><td colspan="2">溶解在500ml的蒸馏水，加​​冰糖麝香草酚。 </td></tr></table><p>冯科萨染色</p><table style="margin-left:40px;" border="0"><tr><td> 1％硝酸银水溶液</td></tr><tr><td> 2.5％硫代硫酸钠</td></tr><tr><td> 1％番红O </td></tr></table><p>阿尔辛蓝染色</p><table style="margin-left:40px;" border="0"><tr><td>阿尔新蓝8GX </td><td> 1克</td></tr><tr><td> 3％醋酸溶液</td><td> 100毫升</td></tr></table><p> Hemotoxylin和曙红染色</p><table style="margin-left:40px;" border="0"><tr><td colspan="2"  style="font-weight:bold;"> A）哈里斯的hemotoxylin </td></tr><tr><td> Hemotoxylin </td><td> 2.5克</td></tr><tr><td>无水酒精</td><td> 25ML </td></tr><tr><td>明矾</td><td> 50克</td></tr><tr><td>蒸馏水</td><td> 500毫升</td></tr><tr><td>氧化汞</td><td> 1.25克或</td></tr><tr><td>碘酸钠</td><td> 0.5克</td></tr><tr><td>冰醋酸</td><td> 20ML </td></tr><tr><td colspan="2"  style="font-weight:bold;"> B）曙红溶液</td></tr><tr><td colspan="2">使用商业化生产的曙红。 </td></tr></table><p>所有染色的染料粉末Gurr染料粉末。 <br />所有试剂均为分析纯，而不是实验室试剂</p><p> <u>荧光</u> </p><table border="1" cellspacing="0" cellpadding="0"><tr><td><br />制造商</td><td>荧光</td><td>目录编号</td></tr><tr><td> Sigma - Aldrich公司，悉尼</td><td>钙黄绿素绿</td><td> C0875 </td></tr><tr><td> Sigma - Aldrich公司，悉尼</td><td>茜素络合剂</td><td> A3882 </td></tr><tr><td>英特威澳大利亚，维多利亚</td><td>土霉素<br /> （现成的抗菌药物） </td><td>商标名称; Engemycin 100 </td></tr></table>

undecalcified bone preparation for histology, histomorphometry and fluorochrome analysis

<p class="jove_content">脱钙骨组织学显示骨微架构。它同时显示矿化骨，它提供了对骨代谢或骨形成和骨吸收的重要信息和细胞成分。这具有巨大的重要性在各种不同的临床和研究中的应用。它会产生美丽的图像技术，如荧光评估和组织形态计量<sup>学2</sup> 1和允许。荧光分析是一种钙结合的荧光染料注射在特定的时间点，在那个特定的时间允许的矿化量的量化技术。组织形态计量学是在微观层面的骨定量过程。 </p><p class="jove_content">执行脱钙骨组织学是在技术上具有挑战性，特别是大尺寸标本。它要求在技术标准石蜡嵌入组织学的变化。该视频演示了质量好的部分生产过程和演示的技术难点和方法，克服这些。实现方式类似于其他软组织的密度和骨的通透性较低，需要相当长的固定和处理时间，但是由于经常服用几个星期，试样制备，固定和处理。嵌入使用类似或等于硬度和密度，如甲基丙烯酸甲酯树脂骨支撑介质，但不像石蜡渗透和嵌入，这是一个不可逆转的步骤。切片可以通过研磨产生较厚的部分，这是最适用于如荧光分析的研究。这是最好的实现上macrotome使用的钻石刀片。另外，较薄，可产生光镜，这是一个非常锋利的刀片使用大锤切片。爬犁切片提供了额外的的大，硬模块所需的强度和稳定性。树脂包埋切片可染各种各样的污渍，这是证明。 </p>

Watch this Scientific Journal Video about Undecalcified Bone Preparation for Histology, Histomorphometry and Fluorochrome Analysis at JoVE.com

Undecalcified Bone Preparation for Histology, Histomorphometry and Fluorochrome Analysis

<p class="jove_content">脱钙骨组织学各种临床和研究中的应用提供了重要信息。这在技术上是具有挑战性，特别是大尺寸的标本。该视频演示了质量好的部分生产过程和演示的技术难点和方法，克服这些。</p>

脱钙骨组织学，形态计量学和荧光染料分析制备

Undecalcified bone histology demonstrates the micro-architecture of bone. It shows both the mineralised and cellular components of bone, which provides ...

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