我们衷心感谢Nanovic研究所欧洲研究，格林家庭荣誉课程，巴黎圣母院综合成像的基金（NDIIF），Carestream Health公司的这个项目的金融支持。支持由美国国家科学基金会BCS-1029149 MJR对兔颅开发研究。

1。动物<ol><li>现将结果报告如下，1的男性Lobund Wistar大鼠10个月的年龄，得到的Freimann的生命科学中心，Notre Dame大学（巴黎圣母院，印第安纳州，美国）。一种离体新西兰白兔（男性，年龄为8周）颅骨样品，保存在10％福尔马林，获得，圣母大学的马修Ravosa教授的实验室。 </li><li> 在活体成像，大鼠麻醉异氟烷（2.5％流量），通过鼻锥体系统的维护。动物俯卧在标准的大鼠床（M2​​M影像公司，美国俄亥俄州克里夫兰市）提供的图像的Albira站。从一个统一的CT收购的躯干四肢被定位横向。 </li><li>图像采集完成后，老鼠从鼻锥的回收笼，直到门诊。 </li><li>对于兔头骨的扫描，将试样置于在大鼠床在一个密封的塑料袋中含有甲醛。 </li></ol> 2。图像采集和重建<ol><li> 在体内和体外进行图像采集使用Albira CT系统（锐珂医疗分子成像，伍德布里奇，CT）的。该系统被设置为扫描通过执行三个圆形扫描（600每次扫描的预测），每个具有一个65毫米的视场角，然后将其缝合在一起，在重建期间，180毫米的长度的床。的X-射线源被设置为400μA和45 kVp的电压的电流，并用0.5毫米的铝过滤器变硬的光束。深约辐射剂量当量CT设置为660毫希，浅层剂量当量为1171毫西弗。这些剂量不超过10倍，低于LD50值。 </li><li>图像重建滤波反投影（FBP）算法，通过使用"标准"的参数的Albira套房5.0重构。这些收购和重建设置产生国际泳联l图像0.125毫米各向同性体素，被视为整体的动物分析和解剖结构的三维打印足够的。 </li></ol> 3。数据处理<ol><li>从CT扫描可能会印从原始数据中没有分割的骨骼特征。然而，软组织分割之前，需要处理数据的三维打印。在这里，我们将展示一个例子，与肺组织。 <ol><li>打开原始的微型正（所有的方式上的Albira的成像系统的数据格式）文件</li><li>周围画一个感兴趣体积（VOI）鼠标，使所有的外部空间被删除。 </li><li>在VOI工具"选项卡上选择"面具外选择VOI和设置屏蔽值-1000菲尔德单位（HU，radiodensity规模CT），这将有效地设置外部空间的空气的CT密度值。 </li><li>删除所有的外部空间，在"工具"选项卡上，选择"外部"，然后点击下拉箭头并选择"分割" </li><li>设定范围为-550至-200，并单击"确定"。 </li><li>如果是非常大的文件，选择"工具"选项卡中，选择"减少"，并运行程序</li><li>分析文件</li></ol></li><li>数据必须先转换成DICOM格式，使用PMOD（PMOD技术有限公司，苏黎世，瑞士）分析软件。 <ol><li>打开PMOD图像处理软件包。 </li><li>除了 ​​排在前列，选择" 查看 "选项卡。 </li><li>在右下角的工具栏上，单击向下箭头标记下的数据库负载 。 </li><li>选择微正为原料的CT数据，或肺的分段分析 。 </li><li>选择相应的文件，然后单击" 添加到收藏栏 。 </li><li>单击" 打开"。 </li><li>在右下角的工具栏中，单击向上箭头下保存 。 </li><li>在此菜单中，选择文件类型DICOM。 </li><li>命名该文件，选择" 保存"。 </li><li>关闭PMOD。 </li></ol></li><li> DICOM数据中包含的每个体素的体积密度值。为了打印此数据，它必须被处理作为一个连续的表面，而不是一个卷。 ImageJ的v1.43u将被用来进行进一步的处理，得到表面渲染。 <ol><li>打开ImageJ的图像处理软件</li><li> 选择File&gt; Import。 </li><li>选择图像序列 。 </li><li>导航到新创建的DICOM文件，其中包含并选择它。 </li><li>选择插件&gt; 3D&gt; 3D浏览器 。 </li><li>会出现两个窗口，3D查看器中会出现一个添加窗口。 </li><li>在显示在添加窗口的变化量表面 。 </li><li> 阈值的默认值更改为210。 </li><li>单击"确定"。 </li><li>在3D查看器菜单栏，选择" 文件"&gt;"导出"&gt;波前表面 。 </li><li>文件命名，然后单击" 保存 "。 </li></ol></li><li> V1.3.1和Meshlab Netfabb工作室基本4.9，两个程序，将同时删除任何多余的网格，加入断开连接的网格，维修孔，并最终网格平滑。这两个方案之间的主要区别是刀具设置提供给用户，和一些接口导航控制。他们都是三维网格编辑软件程序，和他们一起使用，提供了最简单的方法来编辑模型。 <img alt="图标1" fo:src="/files/ftp_upload/50250/50250icon1highres.jpg" src="/files/ftp_upload/50250/50250icon1.jpg" />这些行动表明，应执行中Meshlab V1.3.1 <img alt="图标2" fo:src="/files/ftp_upload/50250/50250icon2highres.jpg" src="/files/ftp_upload/50250/50250icon2.jpg" />表明，这些行动应当执行Netfabb工作室基本4.9 <ol><li>要导入的网格，进入编辑软件: <ol><li>打开Meshlab V1.3.1 <img alt="图标1" fo:src="/files/ftp_upload/50250/50250icon1highres.jpg" src="/files/ftp_upload/50250/50250icon1.jpg" /></li><li>从菜单栏中选择"文件"&gt;"新的空项目 。 </li><li> SELCT" 文件"&gt;"导入"网格 。 </li><li>选择该文件，单击" 打开 "。 </li><li>一旦该文件已加载，一个对话框将打开。保持统一重复的顶点检查，按"确定"。 <ol><li>打开Netfabb工作室基本4.9。 <img alt="图标2" fo:src="/files/ftp_upload/50250/50250icon2highres.jpg" src="/files/ftp_upload/50250/50250icon2.jpg" /></li><li>将所需的文件直接到Netfabb工作室基本画面。 </li></ol></li></ol></li><li>要删除不需要的网格表面: <ol><li>在Meshlab从菜单栏中选择"滤镜"&gt;"&gt;删除隔离件（WRT直径）的清洁和修复 。 <img alt="图标1" fo:src="/files/ftp_upload/50250/50250icon1highres.jpg" src="/files/ftp_upload/50250/50250icon1.jpg" /></li><li>删除孤立的连接组件，其直径小于指定的常数。输入，最大直径为这些组件，然后单击"Apply（应用）。 </li><li>渐渐地，最大直径增加，删除更大的一部分。在每个直径修改后单击" 应用"。 </li></ol></li><li>要加入断开件，孔必须在当前啮合切断一个新的键在所希望的位置，和网状亩的桥梁吨之间建立。 <ol><li>使用选择工具 ，第七工具从左边的菜单栏，选择的网，将切出。 <img alt="图标1" fo:src="/files/ftp_upload/50250/50250icon1highres.jpg" src="/files/ftp_upload/50250/50250icon1.jpg" /></li><li>删除选择删除面"按钮，在菜单栏从左边的第三个工具的网。 </li><li>从菜单栏中， 选择File&gt; Export网作为 。 </li><li>命名该文件，更改文件类型为STL。 </li><li>按" 保存"。 </li><li>会出现一个对话框，保存选项，然后按"确定"。 <img alt="图标2" fo:src="/files/ftp_upload/50250/50250icon2highres.jpg" src="/files/ftp_upload/50250/50250icon2.jpg" /></li><li>将这个新的文件到Netfabb。 </li><li>在顶部的菜单中，选择" 修复"，从左侧的第四个工具。 </li><li>选择" 添加三角形的工具，第十三从左边。 </li><li>点击一侧上的开口边缘（它们将是黄色），然后单击的开口边缘上的另一片上。 5-10桥梁跨越的差距。 </li><li>选择" 自动修复"按钮在右下角</li><li>突出显示默认的修复 。 </li><li>单击" 执行"。 </li><li>按Cancel（ 取消）修复后已被执行。 </li></ol>可选:一些创建的三角形可能是错误的方向。下面的步骤可以用来调整这些三角形的方向。 <ol><li>选择翻转选定的三角形按钮。 </li><li>迷失方向的三角形上任何，旋转三角形在正确的方向。 </li></ol></li><li>对维修坑和洞，整个孔和火山口必须除去桥必须建立在每一侧的网状。 <ol><li>单击" 选择工具 "。 <img alt="图标1" fo:src="/files/ftp_upload/50250/50250icon1highres.jpg" src="/files/ftp_upload/50250/50250icon1.jpg" /></li><li>选择火山口或孔来填补的。 </li><li>单击" 删除面"按钮。 </li><li>从菜单栏中， 选择File&gt; Export网作为 。 </li><li>命名该文件，更改文件类型为STL。 </li><li>按" 保存"。 </li><li>会出现一个对话框，保存选项，然后按"确定"。 </li><li>将这个新的文件到Netfabb。 <img alt="图标2" fo:src="/files/ftp_upload/50250/50250icon2highres.jpg" src="/files/ftp_upload/50250/50250icon2.jpg" /></li><li>在顶部的菜单中，选择" 修复" </st<html>荣&gt;，从左边的第四个工具。 </li><li>选择" 添加三角形的工具，第十三从左边。 </li><li>点击一侧上的开口边缘（它们将是黄色），然后单击的开口边缘上的另一片上。 5-10桥梁跨越的差距。 </li><li>选择" 自动修复"按钮在右下角</li><li>突出显示默认的修复 。 </li><li>单击" 执行"。 </li><li>按Cancel（ 取消）修复后已被执行。 </li></ol></li><li> Laplacian平滑的平滑算法在Meshlab，用于平滑的对象，同时仍然保留模型的结构完整性。 <ol><li>浏览"滤镜"&gt;"平滑&gt;的整流罩和变形&gt;拉普拉斯算子平滑 。iles/ftp_upload/50250/50250icon1.jpg"/&gt; </li><li>选择一个号码的平滑迭代次数。更多的迭代将导致在一个平滑的模型，但每个迭代缓慢降解的音量的模式，这可能会导致在断开连接件和锋利的边缘。建议使用1-5次迭代。 </li><li>按"确定"。 </li><li>从菜单栏中， 选择File&gt; Export网作为 。 </li><li>命名该文件，更改文件类型为STL。 </li><li>按" 保存"。 </li><li>会出现一个对话框，保存选项，然后按"确定"。 </li><li>将这个新的文件到Netfabb。 <img alt="图标2" fo:src="/files/ftp_upload/50250/50250icon2highres.jpg" src="/files/ftp_upload/50250/50250icon2.jpg" /></li><li>在顶部的菜单中，选择" 修复"，从左边的第四个工具。 </li><li>选择" 添加三角形工具，日从左边irteenth。 </li><li>点击一侧上的开口边缘（它们将是黄色），然后单击的开口边缘上的另一片上。 5-10桥梁跨越的差距。 </li><li>选择" 自动修复"按钮在右下角</li><li>突出显示默认的修复 。 </li><li>单击" 执行"。 </li><li>按Cancel（ 取消）修复后已被执行。 </li><li>从菜单栏中， 选择File&gt; Export网作为 。 </li><li>最终的文件命名和更改文件类型为STL。 </li><li>按" 保存"。 </li><li>会出现一个对话框，保存选项，然后按"确定"。 </li></ol></li></ol></li></ol> 4。印刷<ol><li>印刷使用的Makerbot <ol><li>打开STL文件ReplicatorG。复制G是一个沟通的Makerbot Makerbot产业计划。 </li><li>单击" 比例"从右下角菜单，选择填充生成空间 ！ </li><li>选择" 旋转"，然后单击" 平放"。 </li><li>单击中心 。 </li><li> ，细节丰富的模型，:选择填写构建平台 ，以扩展模型。 </li><li>在同一菜单中，单击"上移 "和"选择放在平台 。 </li><li>一旦已经取得了正确的方向，从顶部的菜单栏选择生成GCODE。 会出现一个窗口，打印选项。 </li><li>选择将提供灯丝打印的对象（ 左或右 ）的挤出机。 </li><li>选择使用筏/技术支持 。 </li><li>从下拉菜单中有权我们e支持材料，选择全力支持 。 </li><li>选择" 生成GCODE"。会出现一个弹出框显示进度的GCODE。 </li><li>一旦GCODE已经完成，选择生成文件使用SD卡 。 </li><li>单击" 保存"。 </li><li>拖动文件到SD卡上。 </li><li>将SD卡插入Makerbot使用SD Makerbot键盘选择" 打印 。 </li><li>在打印从SD卡中 ，选择所需的文件名 ​​。 ，Makerbot将自动开始热身打印的对象。 </li></ol></li><li> Shapeways印刷<ol><li>与Shapeways创建一个免费帐户后，STL文件可以直接上传到Shapeways网站: <a href="http://www.shapeways.com/upload/" target="_blank">http://www.shapeways.com/upload/</a> </li><li>点击上传并选择STL文件中。 </li><li>选择一个标题上传的文件。 </li><li>从下拉菜单中选择一个计量单位。 </li><li>点击上传模型 。 </li><li>现在，该文件准备通过Shapeways打印。一旦该文件被上传的Shapeways会花几分钟的时间来处理文件，以确保它实际上可以被打印。大约十分钟后，能打印模式从"我的模式"。 </li><li> "白坚固而有弹性的"选择用于打印骨骼结构，坚固而有弹性的"紫"是用于肺组织。 </li></ol></li><li> PROJET HD 3000印刷<ol><li> STL文件，也可以打印出来使用的是商业高分辨率三维打印机PROJET HD 3000（跳板工程有限责任公司，创业园，巴黎圣母院，IN，美国）等。 </li><li> STL文件被加载到专有软件的3D系统布局工作的platfORM。这需要改变的模型周围的取向，以减少使用的蜡为支撑和打印时间。该文件被保存。 </li><li>然后将该作业以电子方式发送到打印机。 </li><li>铝的平台被加载到打印机和PROJET HD 3000开始打印的对象。 </li><li>模型，然后除去从平台，并放入烘箱，在约73℃以熔化模型的支持蜡。 </li><li>该对象被删除热和擦拭以除去残留的表面蜡与Kimwipe。 </li></ol></li></ol>

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	<li>Kazmer, D. O., Speight, R. G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Polymer+Injection+Molding+Technology+for+the+Next+Millennium.">Polymer Injection Molding Technology for the Next Millennium.</a> Journal of Injection Molding Technology. 1 (2), 81-90 (1997).</li><li>Bradshaw, S., Bowyer, A., Haufe, P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+intellectual+property+implications+of+low-cost+3D+printing.">The intellectual property implications of low-cost 3D printing.</a> ScriptEd. 7 (1), 5-31 (2010).</li><li>Partridge, R., Conlisk, N., Davies, J. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=In-lab+three-dimensional+printing:+An+inexpensive+tool+for+experimentation+and+visualization+for+the+field+of+organogenesis.">In-lab three-dimensional printing: An inexpensive tool for experimentation and visualization for the field of organogenesis.</a> Organogenesis. 8 (1), 1-6 (2012).</li><li>Guillot, A., Champely, S., Batier, C., Thiriet, P., Collet, C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Relationship+between+spatial+abilities,+mental+rotation+and+functional+anatomy+learning.">Relationship between spatial abilities, mental rotation and functional anatomy learning.</a> Adv. Health Sci. Educ. Theory Pract. 12, 491-507 (2007).</li><li>Cignoni, P., Scopigno, R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Sampled+3D+models+for+CH+applications:+a+viable+and+enabling+new+medium+or+just+a+technological+exercise.">Sampled 3D models for CH applications: a viable and enabling new medium or just a technological exercise.</a> Association for Computing Machinery Journal on Computing and Cultural. 1 (1), 1 (2008).</li><li>Symes, M. D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Integrated+3D-printed+reactionware+for+chemical+synthesis+and+analysis.">Integrated 3D-printed reactionware for chemical synthesis and analysis.</a> Nature Chemistry. 4, 349-354 (2012).</li><li>Sheridan, D. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Fabricating+consent:+three-dimensional+objects+as+rhetorical+compositions.">Fabricating consent: three-dimensional objects as rhetorical compositions.</a> Computers and Composition. 27, 249-265 (2010).</li><li>Windisch, G., Salaberger, D., Rosmarin, W., Kastner, J., Exner, G. U., Haldi-Brandle, V., Anderhuber, F. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+Model+for+Clubfood+Based+on+Micro-CT.">A Model for Clubfood Based on Micro-CT.</a> Data. J. Anat. 210, 761-766 (2007).</li></ol>

W.马修Leevy是锐珂医疗分子成像的顾问。布赖恩Stamile是一个支持工程师MakerBot工业。

X射线CT数据集的生活Lobund Wistar大鼠离体新西兰白兔头骨被用来展示3D对象的可行性，从 ​​临床前生产生物数据。模型使用三种不同的来源:1）广受欢迎的Makerbot复制，2）第三方公司Shapeways公司，3）高档商业PROJET HD 3000。每个打印机能够产生满意的原则目标，增强了数据可视化的对象。 在临床前的CT数据的打印过程中，每一个的打印方法的优点和缺点的确定，并总结为最终用户。 MakerBot Replicator是一个便宜的（$ 1,750）台式解决方案，几乎世界各地的任何一个实验室访问。它可以打印多种颜色，用廉价的投入（约3.50元塑料具有润肺老鼠CT）。然而，的Makerbot是有限的分辨率，因此某些机型将适当的挤压和可视化的预期结构，都必须扩大。 Shapeways公司提供了一个优秀的颜色和材料方面的选择。该机型是高分辨率和强大的。虽然它们的价格有关的的MakerBot上每一个单位的基础上（大鼠肺CT与41.61美元）高于10倍，用户可以执行有限数量的就业机会，并避免购买一台打印机的前期费用。两个星期的交货时间从Shapeways是一个小的缺点。 PROJET HD 3000提供了优秀的模型在分辨率和力量。我们很幸运收缩的印刷我们的对象PROJET HD 3000在创新园在巴黎圣母院（约30美元的大鼠肺部CT与劳动力和原材料）。用户可能很难获得这种类型的设备，因为它们的价格在$ 80,000的范围内，它是麻烦的，以及与多种颜色的打印。由于每台仪器/制造商提供了一个不同的度量来描述的分辨率为对象印刷（细节Shapeways最低水平= 0.2毫米，最小壁厚= 0.7毫米，5的 MakerBot片厚度为0.4毫米的喷嘴= 0.2-0.3毫米，6 PROJET HD 3000 DPI = 656所述656×800，精确度为0.025-0.05毫米），各系统之间的相对决议的定性评估显示，Shapeways和的PROJET HD系统，可以打印高细节的规模，为成功地使用某些对象时，必须增大MakerBot。总的来说，所有这三种方法都是环保的，并提供了一​​个方便的方法来实现简便的生产非常详细的临床前研究X射线CT模型。 结论渐渐地，三维打印技术已经被最小化成本和复杂性变得更加方便，8，9，从字面上看任何人都可以打印高分辨率，三维物体从挖ITAL文件。这些详细的三维对象可以是有用的工具，教育工作者和研究人员的一致好评。此外，他们提供的视觉沟通的一种手段，帮助实现更清晰的认识。10例如，医学研究人员可以利用样本或病人的具体型号，以改善与他们的同事和病人的沟通和理解。11虽然在2D屏幕上的表示走过了很长的路，是绝对没有更换的视觉和感官的经验，一个真正的对象，它是能够举行，旋转，审议并移动。配对的电子数据表示模型的功能更强大，因为它允许研究人员，研究的物理对象地区的利益，并在计算机上找到这些领域进一步的定量分析模型。有了适当的数据收集，表面渲染，和stereolit​​hographic编辑，它是能够迅速地生成详细的，relati从X射线CT数据的vely低廉的机型。在这里，我们提供了一个详细的，一步一步的方法用于生产临床前小动物收集的数据与X-射线微CT的三维模型。我们收购了我们的体内和体外 CT数据集使用图像的Albira站PMOD，ImageJ的Meshlab和Netfabb的的套装软件，并进行后续处理。最后，我们提供了一系列的商业解决方案的详细说明，使三维模型打印。在每一种情况下，最终的结果是一个模型，该模型提供了一个独特的，手持式的，物理的体现，所获取的断层图像数据，通常会被限制到一个计算机屏幕。

<table border="1"><tbody><tr><td></td><td></td><td></td><td> 所需的程序 <ol><li> Albira图像获取</li><li> PMOD </li><li> ImageJ的</li><li> Meshlab </li><li> Netfabb </li><li> ReplicatorG </li></ol></td></tr></tbody></table>

3d printing of preclinical x-ray computed tomographic data sets

立体印刷允许非常详细的对象的生产过程通过被称为添加剂制造。传统的模具注射的方法来创建模型或零件有许多限制，其中最重要的是及时的，符合成本效益的方式进行高度复杂的产品中的一个难点。[1]然而，在三维印刷技术的逐步改善，导致在高端和经济工具现在可用于生产的定制机型轻便。这些打印机有能力挤出高分辨率对象有足够的细节，准确地反映在从X射线CT扫描仪的临床前体内生成的图像。有了适当的数据收集，表面渲染，和stereolit​​hographic编辑，它现在是可能的和廉价的从X射线CT数据能够迅速产生详细的骨骼和软组织结构。即使在早期阶段的发展MENT，立体印刷的吸引力都教育工作者和研究人员谁可以利用的技术，以提高可视化能力的解剖模型，3，4的真正好处此方法结果的有形经验的研究员可以有数据不能被充分转达了通过电脑屏幕上。临床前的三维数据的物理对象，是一份精确的测试对象是一个功能强大的可视化和通信工具，特别是对涉及成像研究的学生，或那些在其他领域的翻译。在这里，我们提供了一个详细的方法用于印刷塑料模型的骨骼和器官的结构，来自于X射线CT扫描利用一个Albira在PMOD ImageJ的，Meshlab，Netfabb结合的X射线CT系统，和ReplicatorG软件包的。

<img alt="图1" src="/files/ftp_upload/50250/50250fig1.jpg" /> 图1。三维打印模型大鼠X射线CT数据集的肺部和骨骼特征。对象使用一个PROJET HD 3000（左），Shapeways公司（中心）或一个:Makerbot复制（右）打印。比例尺表示2厘米。请注意，在面板C的比例尺是小于A和B，这反映Makerbot必须打印在某些情况下，为了输出足够的细节的放大的对象。 图1描绘了三个打印相同的方法在大鼠体内的CT数据集的最终产品。所有这三种型号包括裁剪后的骨骼结构和可移动的肺部独立打印和拼凑起来的。在左边的模型是PROJET HD 3000的高分辨率打印机，使用半透明的丙烯酸塑料的结果。中心处的对象使用第三方公司，Shapeways公司，它的骨骼结构，使用尼龙12的白色塑料印刷，而对呼吸系统的结构是捏造的紫色。这些前两种模式打印的实际规模，测量长度约11厘米。在右边的对象是使用MakerBot。上述骨骼构造的印刷使用天然颜色的ABS（丙烯腈 - 丁二烯 - 苯乙烯）塑料和肺与石灰绿色ABS。由于分辨率的限制的MakerBot，这种模式可以不被打印，而不会降低性能的精细结构，如肋骨规模。相反，这个模型是由几乎2X缩放使用"填充建立空间"选项，以获得所需的视觉细节，从而在21厘米的长度的一个目的。 <img alt="图2" src="/files/ftp_upload/50250/50250fig2.jpg" /> 图2。三维打印模型的离体兔SKULL的数据集。打印显示的对象使用一个PROJET HD 3000（左），:Shapeways有限公司（中心）和一个:Makerbot复制（右）。比例尺表示1厘米。 图2示出了每个体外兔颅骨CT数据集的打印方法的最终产品。在左边的模型是PROJET HD 3000的高分辨率打印机使用半透明的丙烯酸塑料制成的结果。该模型通过在中心被印在白色nylon12塑料Shapeways印刷。在右边的对象被印在白色的塑料使用MakerBot。所有这三个对象被打印到规模和测量长度约8.5厘米。 <img alt="图3" src="/files/ftp_upload/50250/50250fig3.jpg" /> 图3。 3D打印模式，一个完整的大鼠X射线CT数据集对象使用一个PROJET HD 3000（左）和:Shapeways股份有限公司（右）打印。刻度尺b芳表示1厘米。 图3示出印刷的一个完整的大鼠体内 CT数据集的两个方法的最终产品。这两个模型包含了一个完整的骨骼结构（减去尾巴）和可移动的肺部。在左边的模型是一个高分辨率的打印机，PROJET HD 3000使用半透明的丙烯酸类塑料，印刷所得到的。该模型在正确的Shapeways印刷，印刷用使用白色nylon12塑料和肺部的紫色创建的骨骼结构。这两款车型被打印到实际的规模，测量长度约19厘米。由于需要复杂的细节，不能被打印的完整骨架的MakerBot复制。 在三维印刷技术的勘探，观察到一定的优点和缺点，并概述在表1中。 <table border="1"><tbody><tr><td><stroNG&gt;印刷方法</td><td> 优点 </td><td> 缺点 </td></tr><tr><td> MakerBot </td><td>速度非常快，多种颜色可供选择，能够打印两种颜色可供选择，非常便宜</td><td>最低级别的详细信息。去除载体材料是缓慢的（在几个小时的顺序）。 </td></tr><tr><td> Shapeways </td><td>色彩选择Varity，各种材料的印刷，高层次的细节，价格相对低廉</td><td>两个星期的时间来处理和接收订单</td></tr><tr><td> PROJET HD 3000 </td><td>相对快速周转的细节，层次最高，高吞吐量，容易清除支持材料（蜡）。 </td><td>最昂贵的高达前期成本，在实际使用的只有一个颜色选项。 </td></tr></tbody></table> 表1中。对照三维打印技术，可打印CT数据集。

Watch this Scientific Journal Video about 3D Printing of Preclinical X-ray Computed Tomographic Data Sets at JoVE.com

3D Printing of Preclinical X-ray Computed Tomographic Data Sets

使用现代塑料挤出和印刷技术，现在有可能快速且廉价的透视拍摄的CT数据在实验室中产生的物理模型。断层数据的三维印刷是一个功能强大的可视化，研究和教育工具，现在可以访问的临床前成像社区。

基础X射线计算机断层数据的三维打印设置

Three-dimensional printing allows for the production of highly detailed objects through a process known as additive manufacturing.  Traditional, ...

3d-printing-of-preclinical-x-ray-computed-tomographic-data-sets

Research

JoVE Journal

Biology

40.3K 次观看.  University of Notre Dame. 使用现代塑料挤出和印刷技术，现在有可能快速且廉价的透视拍摄的CT数据在实验室中产生的物理模型。断层数据的三维印刷是一个功能强大的可视化，研究和教育工具，现在可以访问的临床前成像社区。

视频: 3D Printing of Preclinical X-ray Computed Tomographic Data Sets

Non-invasive 3D-Visualization with Sub-micron Resolution Using Synchrotron-X-ray-tomography

Little is known about the internal organization of many micro-arthropods with body sizes below 1 mm. The reasons for that are the small size and the hard cuticle which makes it difficult to use protocols of classical histology. In addition, histological sectioning destroys the sample and can therefore not be used for unique material. Hence, a non-destructive method is desirable which allows to view inside small samples without the need of sectioning.
We used synchrotron X-ray tomography at the European Synchrotron Radiation Facility (ESRF) in Grenoble (France) to non-invasively produce 3D tomographic datasets with a pixel-resolution of 0.7&micro;m. Using volume rendering software, this allows us to reconstruct the internal organization in its natural state without the artefacts produced by histological sectioning. These date can be used for quantitative morphology, landmarks, or for the visualization of animated movies to understand the structure of hidden body parts and to follow complete organ systems or tissues through the samples.

We used synchrotron X-ray tomography at the European Synchrotron Radiation Facility (ESRF) to non-invasively produce 3D tomographic datasets with a pixel-resolution of 0.7&micro;m. Using volume rendering software, this allows the reconstruction of internal structures in their natural state without the artefacts produced by histological sectioning.

非侵入性的三维可视化与亚微米分辨率使用同步加速器X射线断层扫描

Little is known about the internal organization of many micro-arthropods with body sizes below 1 mm. The reasons for that are the small size and the hard ...

科研

生物学

Microcontact Printing of Proteins for Cell Biology

The ability to pattern proteins and other biomolecules onto substrates is important for capturing the spatial complexity of the extracellular environment. Development of microcontact printing by the Whitesides group (<a href="http://gmwgroup.harvard.edu/">http://gmwgroup.harvard.edu/</a>) in the mid-1990s revolutionalized this field by making microelectronics/microfabrication techniques accessible to laboratories focused on the life sciences. Initial implementations of this method used polydimethylsiloxane (PDMS) stamps to create patterns of functionalized chemicals on material surfaces1. Since then, a range of innovative approaches have been developed to pattern other molecules, including proteins2. This video demonstrates the basic process of creating PDMS stamps and uses them to pattern proteins, as these steps are difficult to accurately express in words. We focus on patterning the extracellular matrix protein fibronectin onto glass coverslips as a specific example of patterning.
An important component of the microcontact printing process is a topological master, from which the stamps are cast; the raised and lowered regions of the master are mirrored into the stamp and define the final pattern. Typically, a master consists of a silicon wafer coated with photoresist and then patterned by photolithography, as is done here. Creation of masters containing a specific pattern requires specialized equipment, and is best approached in consultation with a fabrication center or facility. However, almost any substrate with topology can be used as a master, such as plastic diffraction gratings (see Reagents for one example), and such serendipitous masters provide readily available, simple patterns. This protocol begins at the point of having a master in hand.

Microcontact printing is used extensively to pattern proteins and other molecules on material surfaces. We demonstrate the basic steps of this process, stamping patterns of fibronectin onto glass.

微接触印刷的蛋白细胞生物学

The ability to pattern proteins and other biomolecules onto substrates is important for capturing the spatial complexity of the extracellular environment. ...

Protein Crystallization for X-ray Crystallography

Using the three-dimensional structure of biological macromolecules to infer how they function is one of the most important fields of modern biology. The availability of atomic resolution structures provides a deep and unique understanding of protein function, and helps to unravel the inner workings of the living cell. To date, 86% of the Protein Data Bank (rcsb-PDB) entries are macromolecular structures that were determined using X-ray crystallography.
To obtain crystals suitable for crystallographic studies, the macromolecule (e.g. protein, nucleic acid, protein-protein complex or protein-nucleic acid complex) must be purified to homogeneity, or as close as possible to homogeneity. The homogeneity of the preparation is a key factor in obtaining crystals that diffract to high resolution (Bergfors, 1999; McPherson, 1999).
Crystallization requires bringing the macromolecule to supersaturation. The sample should therefore be concentrated to the highest possible concentration without causing aggregation or precipitation of the macromolecule (usually 2-50 mg/ mL). Introducing the sample to precipitating agent can promote the nucleation of protein crystals in the solution, which can result in large three-dimensional crystals growing from the solution. There are two main techniques to obtain crystals: vapor diffusion and batch crystallization. In vapor diffusion, a drop containing a mixture of precipitant and protein solutions is sealed in a chamber with pure precipitant. Water vapor then diffuses out of the drop until the osmolarity of the drop and the precipitant are equal (Figure 1A). The dehydration of the drop causes a slow concentration of both protein and precipitant until equilibrium is achieved, ideally in the crystal nucleation zone of the phase diagram. The batch method relies on bringing the protein directly into the nucleation zone by mixing protein with the appropriate amount of precipitant (Figure 1B). This method is usually performed under a paraffin/mineral oil mixture to prevent the diffusion of water out of the drop.
Here we will demonstrate two kinds of experimental setup for vapor diffusion, hanging drop and sitting drop, in addition to batch crystallization under oil.

The 3-D structure of a molecule provides a unique understanding of how the molecule functions. The principal method for structure determination at near-atomic resolution is X-ray crystallography. Here, we demonstrate the current methods for obtaining three-dimensional crystals of any given macromolecule that are suitable for structure determination by X-ray crystallography.

X射线晶体学的蛋白质结晶

Using the three-dimensional structure of biological macromolecules to infer how they function is one of the most important fields of modern biology. The ...

Do's and Don'ts of Cryo-electron Microscopy: A Primer on Sample Preparation and High Quality Data Collection for Macromolecular 3D Reconstruction

Cryo-electron microscopy (cryoEM) entails flash-freezing a thin layer of sample on a support, and then visualizing the sample in its frozen hydrated state by transmission electron microscopy (TEM). This can be achieved with very low quantity of protein and in the buffer of choice, without the use of any stain, which is very useful to determine structure-function correlations of macromolecules. When combined with single-particle image processing, the technique has found widespread usefulness for 3D structural determination of purified macromolecules. 
The protocol presented here explains how to perform cryoEM and examines the causes of most commonly encountered problems for rational troubleshooting; following all these steps should lead to acquisition of high quality cryoEM images. The technique requires access to the electron microscope instrument and to a vitrification device. Knowledge of the 3D reconstruction concepts and software is also needed for computerized image processing. Importantly, high quality results depend on finding the right purification conditions leading to a uniform population of structurally intact macromolecules. 
The ability of cryoEM to visualize macromolecules combined with the versatility of single particle image processing has proven very successful for structural determination of large proteins and macromolecular machines in their near-native state, identification of their multiple components by 3D difference mapping, and creation of pseudo-atomic structures by docking of x-ray structures. The relentless development of cryoEM instrumentation and image processing techniques for the last 30 years has resulted in the possibility to generate de novo 3D reconstructions at atomic resolution level.

This paper describes the cryo-electron microscopy methodology, which is used to obtain high quality microscopic images of macromolecules in their near-native state. The method yields images suitable for further computerized processing using the single particle approach, devised to generate the 3D reconstruction of a macromolecule.

该做什么和不该做什么低温电子显微镜的:入门样品制备和高质量的数据采集为大分子三维重建

Cryo-electron microscopy (cryoEM) entails flash-freezing a thin layer of sample on a support, and then visualizing the sample in its frozen hydrated state ...

SpOT the Correct Tissue Every Time in Multi-tissue Blocks

Multi-tissue paraffin blocks provide high throughput analysis with increased efficiency, experimental uniformity, and reduced time and cost. Tissue microarrays make up the majority of multi-tissue paraffin blocks, but increasingly, researchers are using non-arrayed blocks containing larger tissues from multiple individuals which can provide many of the advantages of tissue microarrays without substantial investment in planning and equipment. A critical component of any multi-tissue analysis is the orientation method used to identify each individual tissue. Although methods exist to maintain proper orientation and identification of tissues in multi-tissue blocks, most are not well-suited to non-arrayed blocks, may consume valuable space within an array and/or are difficult to produce in the standard histology laboratory. The Specimen Orientation Tag (SpOT) is a simple, low cost orientation tool that is clearly visible in paraffin blocks and all tissue sections for reliable specimen identification in arrayed and non-arrayed layouts. The SpOT provides advantages over existing orientation methods for non-arrayed blocks as it does not require any direct modification to the tissue and allows for flexibility in the arrangement of tissue pieces.

The purpose of the Specimen Orientation Tag (SpOT) is to function as an orientation tool to aid in individual tissue identification in multi-tissue paraffin blocks. These protocols demonstrate how it is constructed easily from common, low-cost histology materials and serves as a reliable visual marker in paraffin blocks and sections.

现货正确的组织多组织块每一次

Multi-tissue paraffin blocks provide high throughput analysis with increased efficiency, experimental uniformity, and reduced time and cost. Tissue ...

Production of Human Norovirus Protruding Domains in E. coli for X-ray Crystallography

The norovirus capsid is composed of a single major structural protein, termed VP1. VP1 is subdivided into a shell (S) domain and a protruding (P) domain. The S domain forms a contiguous scaffold around the viral RNA, whereas the P domain forms viral spikes on the S domain and contains determinants for antigenicity and host-cell interactions. The P domain binds carbohydrate structures, i.e., histo-blood group antigens, which are thought to be important for norovirus infections. In this protocol, we describe a method for producing high quality norovirus P domains in high yields. These proteins can then be used for X-ray crystallography and ELISA in order to study antigenicity and host-cell interactions.
The P domain is firstly cloned into an expression vector and then expressed in bacteria. The protein is purified using three steps that involve immobilized metal-ion affinity chromatography and size exclusion chromatography. In principle, it is possible to clone, express, purify, and crystallize proteins in less than four weeks, which makes this protocol a rapid system for analyzing newly emerging norovirus strains.

Production of Human Norovirus Protruding Domains in E. coli for X-ray Crystallography

Here, we describe a method to express and purify high quality norovirus protruding (P) domains in E. coli for use in X-ray crystallography studies. This method can be applied to other calicivirus P domains, as well as non-structural proteins, i.e., viral protein genome-linked (VPg), protease, and RNA dependent RNA polymerase (RdRp).

人类诺沃克病毒凸，域的生产<em&gt;电子。大肠杆菌</em&gt; X射线晶体学

The norovirus capsid is composed of a single major structural protein, termed VP1. VP1 is subdivided into a shell (S) domain and a protruding (P) domain. ...

High-throughput Measurement of Gut Transit Time Using Larval Zebrafish

Zebrafish are used as alternative model organisms for drug safety testing. The gastrointestinal (GI) tract of zebrafish has genetic, neuronal, and pharmacological similarities to that of the mammals. GI intolerance during clinical testing of drug candidates is common and may pose a serious threat to human health. Testing for GI toxicity in preclinical mammalian models can be expensive in terms of time, test compound, and labor. The high-throughput method presented here may be used to predict GI safety issues. Compared to mammalian models, this method allows for more expedient assessment of test compound effects on GI transit while using low quantities of compound. In this method, larval zebrafish (7 days post fertilization) are fed food containing a fluorescent label. After feeding, each larval fish is placed into a well of a 96-conical-bottom-well plate and dosed with test compound (dissolved in water) or the vehicle. As gut transit occurs, fecal matter accumulates on the bottom of the wells, and the rate at which this happens is monitored by measuring fluorescence from the bottom of the well repeatedly over time using a plate spectrophotometer. The fluorescence from larvae in a given treatment group are averaged and these values are graphed along with standard error, for each measurement time, yielding a curve representing average transit of food over time. Effects on gut transit time are identified by comparing the area under the curve for each treatment group to that of the vehicle-treated group. This method detected changes in zebrafish GI transit time induced by drugs with known clinical GI effects; it can be employed to interrogate dozens of treatments for GI effects per day. As such, safer compounds can be quickly prioritized for mammalian testing, which expedites discovery and proffers 3Rs advancement.

The goal of this protocol is to measure the transit time of fluorescently labeled food through the gut of larval zebrafish in a high throughput fashion.

利用斑马鱼对肠道转运时间进行高通量测量

Zebrafish are used as alternative model organisms for drug safety testing. The gastrointestinal (GI) tract of zebrafish has genetic, neuronal, and ...

Micron-scale Phenotyping Techniques of Maize Vascular Bundles Based on X-ray Microcomputed Tomography

It is necessary to accurately quantify the anatomical structures of maize materials based on high-throughput image analysis techniques. Here, we provide a 'sample preparation protocol' for maize materials (i.e., stem, leaf, and root) suitable for ordinary microcomputed tomography (micro-CT) scanning. Based on high-resolution CT images of maize stem, leaf, and root, we describe two protocols for the phenotypic analysis of vascular bundles: (1) based on the CT image of maize stem and leaf, we developed a specific image analysis pipeline to automatically extract 31 and 33 phenotypic traits of vascular bundles; (2) based on the CT image series of maize root, we set up an image processing scheme for the three-dimensional (3-D) segmentation of metaxylem vessels, and extracted two-dimensional (2-D) and 3-D phenotypic traits, such as volume, surface area of metaxylem vessels, etc. Compared with traditional manual measurement of vascular bundles of maize materials, the proposed protocols significantly improve the efficiency and accuracy of micron-scale phenotypic quantification.

We provide a novel method to improve the X-ray absorption contrast of maize tissue suitable for ordinary microcomputed tomography scanning. Based on CT images, we introduce a set of image-processing workflows for different maize materials to effectively extract microscopic phenotypes of vascular bundles of maize.

基于 X 射线 Microcomputed 层析成像的玉米血管束微米尺度分型技术研究

It is necessary to accurately quantify the anatomical structures of maize materials based on high-throughput image analysis techniques. Here, we provide a ...

Dosimetry for Cell Irradiation using Orthovoltage (40-300 kV) X-Ray Facilities

The importance of dosimetry protocols and standards for radiobiological studies is self-evident. Several protocols have been proposed for dose determination using low energy X-ray facilities, but depending on the irradiation configurations, samples, materials or beam quality, it is sometimes difficult to know which protocol is the most appropriate to employ. We, therefore, propose a dosimetry protocol for cell irradiations using low energy X-ray facility. The aim of this method is to perform the dose estimation at the level of the cell monolayer to make it as close as possible to real cell irradiation conditions. The different steps of the protocol are as follows: determination of the irradiation parameters (high voltage, intensity, cell container etc.), determination of the beam quality index (high voltage-half value layer couple), dose rate measurement with ionization chamber calibrated in air kerma conditions, quantification of the attenuation and scattering of the cell culture medium with EBT3 radiochromic films, and determination of the dose rate at the cellular level. This methodology must be performed for each new cell irradiation configuration as the modification of only one parameter can strongly impact the real dose deposition at the level of the cell monolayer, particularly involving low energy X-rays.

Dosimetry for Cell Irradiation using Orthovoltage 40-300 kV X-Ray Facilities

This document describes a new dosimetry protocol for cell irradiations using low energy X-ray equipment. Measurements are performed in conditions simulating real cell irradiation conditions as much as possible.

使用矫形器（40-300 kV）X射线设施进行细胞辐照的测定

The importance of dosimetry protocols and standards for radiobiological studies is self-evident. Several protocols have been proposed for dose ...

A 3D Cartographic Description of the Cell by Cryo Soft X-ray Tomography

Imaging techniques are fundamental in order to understand cell organization and machinery in biological research and the related fields. Among these techniques, cryo soft X-ray tomography (SXT) allows imaging whole cryo-preserved cells in the water window X-ray energy range (284-543 eV), in which carbon structures have intrinsically higher absorption than water, allowing the 3D reconstruction of the linear absorption coefficient of the material contained in each voxel. Quantitative structural information at the level of whole cells up to 10 &#181;m thick is then achievable this way, with high throughput and spatial resolution down to 25-30 nm half-pitch. Cryo-SXT has proven itself relevant to current biomedical research, providing 3D information on cellular infection processes (virus, bacteria, or parasites), morphological changes due to diseases (such as recessive genetic diseases) and helping us understand drug action at the cellular level, or locating specific structures in the 3D cellular environment. In addition, by taking advantage of the tunable wavelength at synchrotron facilities, spectro-microscopy or its 3D counterpart, spectro-tomography, can also be used to image and quantify specific elements in the cell, such as calcium in biomineralization processes. Cryo-SXT provides complementary information to other biological imaging techniques such as electron microscopy, X-ray fluorescence or visible light fluorescence, and is generally used as a partner method for 2D or 3D correlative imaging at cryogenic conditions in order to link function, location, and morphology.

Here, a protocol describing the sample preparation and data collection steps required in cryo soft X-ray tomography (SXT) to image the ultrastructure of whole cryo-preserved cells at a resolution of 25 nm half pitch, is presented.

通过冷冻软X射线断层扫描对细胞进行3D制图描述

Imaging techniques are fundamental in order to understand cell organization and machinery in biological research and the related fields. Among these ...