作者感谢牙科技师尼古拉Lanfranconi为他们与改善扫描软件的持续支持生产主参考模型和Alicona公司。

1，试样制备/基地<ol><li>适用围绕标本平底。将试样在扫描表。定向的牙合面的水平面（ 图1）。 </li></ol> 2，软件分析<ol><li>启动该程序，然后在实验室测量模块（ 图2） </li><li>放置标本的扫描表的中心位置。 </li><li>选择正确的倍率镜头。对于大的对象，如满弓的扫描，使用5倍的目标。 </li><li>通过使用三维鼠标，直到试样的表面被显示在实时取景窗口（ 图3）上移动的扫描光学器件。 </li><li>使用传感器控制来设定曝光和对比度，以达到最佳的扫描参数。用于金属表面，用400和800微秒至0.3和0.8之间的对比度之间的曝光（ 图4）。 </li><li>检查图像质量通过检查按钮"显示图像质量的实时预览"。 </li><li>在部分"测量控制"设置正确参数。测量类型是"3D数据集"; ImageField的类型是"常规的ImageField"（ 图5）。 </li><li>单击"新建的ImageField"。 </li><li>定义扫描体积。该软件需要测量体积的边界坐标。 <ol><li>将试样的最高和最低的扫描级别，然后单击"添加位置"在这两点。在"Z系列"值显示扫描容积的实际高度（ 图6）。 </li><li>将试样转移到扫描体积的XY boundings。点击"添加位置"来定义X轴和Y轴的长度。在扫描体积的尺寸显示在"信息"选项卡，并应该超过试样的尺寸由1厘米的X轴和Y轴。 </li></ol></li><li>勾选"点"数。该软件能够扫描亿面点合一"的ImageField"扫描。实际数目点超过此限制。 "抽取"的测量点的数量由点大小的"横向缩减像素采样"。 <ol><li>点击"高级设置"（ 如图7）。 </li><li>将"横向缩减像素采样"滑块到右边，直到"点"人数减少至低于100米（ 图7）。下采样减少了表面的点的横向分辨率，从而产生被扫描物体的更大的像素大小。为了确保最佳的扫描结果，垂直分辨率增加。 </li></ol></li><li>点击"开始测试"。这将启动"预览模式"。该软件执行预扫描与所选择的X-和Y-轴尺寸。 </li><li>预扫描完成后，选择感兴趣的区域。这有助于减少文件大小和扫描时间（ 图8）。 </li><li>选择预扫描的所有部分以外的标本和两个度量字段周围的样品，将含有扁平基座（ 图9）。 </li><li>点击"开始"按钮开始扫描。 </li><li>控制扫描中使用鼠标在"图像浏览器"，然后按鼠标左键（ 图10）。 </li><li>关闭窗口，点击"查看伪彩色只"。 </li><li>点击"设置"和"伪着色"，然后选择"重复"。 </li><li>设置"最大。"重视到0.2，然后点击"应用范围"。 </li><li>控制的可重复性，它应该等于具有相同的倾斜和材料领域。尤其是，周围的样品基体应显示均匀的可重复性（ 图11）。 </li><li>单击"数据库"和扫描保存到相应的文件夹。 </li><li>扫描导出到不同的文件格式，如果需要的话。点击"F​​ile/Export/3D数据作为/ ...."该协议可以在该点暂停，稍后继续。 </li></ol> 3。差异分析<ol><li>使用"3D编辑器"来切去的基础。这个区域将不被用于区别分析。 </li><li>比较和分析两次扫描，启动"差测量"软件（ 图12）。 </li><li>选择第二个模型进行比较。 </li><li>点击"自动粗糙对齐"执行第一的车型匹配。 </li><li>点击"手动调整"和对准模型旋转和移动，直到它们在相同的方向（ 图13）。 </li><li>点击"自动对齐"和"应用"开始最合适算法的最优模型匹配。 </li><li>点击"差异"来查看两个匹配模型的差异图。 </li><li>选择一种合适的颜色范围以显示偏差（ 图14）。除视觉偏差图案作为屏幕截图进行分析。 </li></ol>点击"统计"显示的不同的统计值。选择1微米级的尺寸和直方图数据保存到一个文本文件进行统计对比（ 图15）。

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作者宣称，他们有没有竞争的经济利益或其他利益冲突。

精度是在牙科医学的一个基本要求。参考扫描器能够扫描小型和大型物体的高正确度与精密度的。用最佳的扫描方法中，即使个别形态详细牙齿表面可以被扫描以高的分辨率和可重复性。随着扫描仪的不同放大倍数下，宏观和微观形态结构可以被收购。它可以扫描各种模型材料。 参考扫描仪的精度评估包括不同型号的取向和相同的模型方位揭示扫描和软件滤波算法12重复扫描的扫描。以从患者的印象衍生的不锈钢全弓模型进行了精度测量，以验证该参考扫描器。本程序，在扫描过程本身的偏差进行定量和显示的值正确度与精密度不扫描形态的先验知识参考扫描器的。与此相反，坐标测量机，或CMM测量设备，也可用于参考测量值，不能扫描，因为tipball的大小的详细的牙齿结构。有了这样的测量设备，只有几面点和特殊的几何形状可以捕获15。 作为一个优势，扫描软件，能够显示每幅扫描表面点的可重复性。低可重复性是指表面点的较低的扫描精度。这不仅会影响当地的精度，而且整个扫描对象的全局精度为单一图像扫描后缝合在一起。有了这个功能，扫描参数的优化选择，可以确保最佳的扫描效果。此外，该扫描仪是可调的，以一个巨大的各种扫描材料。粗糙的表面可以被扫描与普通光的反射和调整对比度和亮度。表面具有高反射性能， 如抛光的金属可以与偏振滤光器进行扫描，以避免不规则的反射，得到一个干净的测量。对于具有陡峭的倾斜对象，环形灯最好照亮这些地区与这个额外的光源。 牙齿表面的特产是形态。与陡峭的斜坡，以口头和颊侧标本结束。这些地区以低可重复性导致较低的精度，拼接大对象在一起，就像全牙弓扫描时尤其如此。以确保充分的牙弓的扫描精度高，有必要环绕​​该模型具有平坦基座。基材必须具有相同的扫描参数作为模型材料。 与该扫描方法，它有可能用相同的评价方法进行比较常规和数字曝光，并给予一个目录ECT比较。表面与差分图像的三维比较有助于确定每印象的方法的具体错误，并可以用来改进的印象的通过开发一个最佳印象程序的质量。迄今为止，牙科扫描仪的验证只被使用的小测试对象11描述。 在（90-10）/ 2个百分点，用于描述两个表面的平均偏差。这个值描述了80％的从基准模型​​中的测试对象的表面的最大距离。表面的最高和最低的10％不采取因型号12的利润率的影响，不同的扫描尺寸考虑在内。 参考扫描器的限制是扫描时间。重要结构和湿样品不​​能直接扫描用此方法。另一个限制是固定的扫描方向。与单一扫描，削弱canno吨被扫描。从不同方向的扫描可以被匹配到扩展模型。像无任何填料含量的树脂材料的非结构材料不能以高精确度，因为它们的低反射特性的扫描。这些材料需要的表面涂层， 例如金溅射。 目前的结果表明，该参考扫描器评价印象的准确性有很大的好处。三维对比在许多研究3,6,7使用的简单的二维测量距离相比提供了更多的信息。不同的印象的方法和变形的原点的局部效应可以被评估，尤其是与数字印象的方法13。填充材料的磨损稳定性可以用真正的牙齿形态无几何限制进行评估。将来的利息字段将使用牙齿咬合，牙齿修复的配合，以及粗糙度measurem的精度计算机辅助设计/计算机辅助制造（CAD / CAM）铣削材料后已废除。

精度是在口腔医学领域的许多的重大利益。更换牙体硬组织需要一个确切的装配假肢，以确保适当的功能，防止进一步破坏剩余的牙体组织1,2。固定义齿和全假体是精确拟合在像备牙或种植体3支撑结构尤为重要。这就是为什么一个高度精确的再现是必要的，尤其是在牙齿印模和牙科实验室的工作流的领域。然而，牙科治疗等领域也受益于一个真实和精确测量的结果，以确认治疗成功和评估新的治疗策略， 如软，硬组织填充，侵蚀和磨损监控，牙周治疗和正畸治疗4,5。在许多领域，目前的验证过程是直线距离的测量用游标卡尺或显微镜6,7。这些甲基ODS仅限于几个测量点和测试区域的三维（3D）的变化的信息有限。较新的测量方法包括在测试对象8,9的整个表面上的光或放射线捕捉。这里，在整个表面或体积的测定和显示在计算机屏幕上的3D对象。线性测量是可能的，以及模型从不同的扫描时间叠印。与此叠加，在每一个扫描点的表面变化的评估是可能的。这使得能够监控特定区域或所有三个坐标轴显示变形的可能性。另外，体积的变化可以测量10。限制点与这些新的方法是在扫描仪的精确度，用于捕获被检测物。没有参考扫描器的精度范围内的变化可分为测试对象或扫描误差的变化。扫描精度通常是由马努给出的值造商来自扫描小，校准对象11。扫描大型物体像一个牙弓时，这个最小的扫描误差是不同的。精度是由正确度和精确度。真实性是被扫描物体的从它的实际几何形状的偏差。精度是重复扫描（ISO 5725-1）之间的偏差。在这项研究中，一种新的光学参考扫描器的基础上，重点变异扫描技术，介绍了从单一的牙齿标本扫描到全拱模型精度最高。此参考扫描器被用作一些研究基地，从常规和数字技术12-14和对有关牙科咬合和牙科材料的磨损实际项目比较牙齿印模的精度。本研究的目的是为了提供参考扫描器的精度和基本信息的一些可能性，在牙科领域的研究使用该设备。

<table border="0" cellpadding="0" cellspacing="0" width="669">
	<tbody>
		<tr>
			<td>Reference model</td>
			<td></td>
			<td></td>
			<td>individual non-precious metal model, derived from a patient impression</td>
		</tr>
		<tr>
			<td>Araldit repair</td>
			<td>Huntsmen Advanced Material, Basel, Switzerland</td>
			<td></td>
			<td>used for making the base of the reference model</td>
		</tr>
		<tr>
			<td>CamBase</td>
			<td>Dentona, Dortmund, Germany</td>
			<td></td>
			<td>Type IV dental ston for pouring conventional impressions</td>
		</tr>
		<tr>
			<td>Identium</td>
			<td>Kettenbach, Eschenburg, Germany</td>
			<td></td>
			<td>Vinylsiloxanether impression material for conventional impression</td>
		</tr>
		<tr>
			<td>inEOS model holder</td>
			<td>Sirona Dental Systems, Bensheim, Germany</td>
			<td></td>
			<td>used for fixing stone models at the reference scanner</td>
		</tr>
		<tr>
			<td>Accutrans</td>
			<td>Coltene Whaledent, Altst&#228;tten, Switzerland</td>
			<td></td>
			<td>used for making the base of thestone models</td>
		</tr>
		<tr>
			<td>President putty</td>
			<td>Coltene Whaledent, Altst&#228;tten, Switzerland</td>
			<td></td>
			<td>mix with accutrans for betterstability of the base</td>
		</tr>
		<tr>
			<td>Alicona Infinite Focus</td>
			<td>Alicona Imaging, Graz, Austria</td>
			<td></td>
			<td>Reference scanner&#160;</td>
		</tr>
	</tbody>
</table>

accuracy in dental medicine, a new way to measure trueness and precision

参考扫描仪是用于牙科医学验证了很多程序。其主要兴趣是验证方法的印象，因为他们作为牙齿修复基地。许多参考扫描仪目前的限制是缺乏准确的扫描大型物体，如全牙弓，或在有限的可能性，以评估具体的牙齿表面。一种新的参考扫描器的基础上，重点变异扫描技术，是与问候最高的局部和全身的准确性进行评估。一个具体的扫描协议进行了测试，从牙齿印模扫描原始牙齿表面。此外，不同型号的材料进行了核实。结果表明:该参考扫描器的高扫描精度为5.3±1.1微米为正确度的平均偏差和1.6±0.6微米的精密在箱子满拱扫描。目前的牙齿印模的方法表现出更高的偏差（正确度:20.4±2.2微米，精度:12.5±2.5微米）笔韩参考扫描器内部扫描精度。像单齿面较小的对象可以与一个更高的精度进行扫描，使系统能够评估腐蚀性和磨蚀牙齿表面的损失。参考扫描器可用于测量差异进行了大量的牙科研究领域。在不同的放大倍数水平结合了高的局部和全身的精度，可用于评估的单齿或修复体达到充分拱的变化的变化。

图16A示出了石膏样品的扫描。为了验证最佳的扫描参数，控制重复性的材料。试样的部分进行扫描具有不同的对比度和亮度设置，在每次扫描后的可重复性进行检查以找到最佳的扫描设置。金属表面显示出与对比低1.0和石膏或树脂材料具有更高的对比度达到最佳效果。 图16B示出了测试的扫描之后没有最佳的扫描参数的可重复性。的倾斜尖表面显示出低的可重复性。 图16C示出了同样的表面扫描具有最佳对比度和亮度。 图17A示出了牙弓的扫描。在陡峭的斜坡在颊和口腔方面透露了一个低重复性（ 图17B）。这种扫描的真实性被显示在图17C。平底，周围的型号，significantly增加扫描精度，包括在扫描（ 图17D-17F）的时候。 图18显示了研究协议来检查扫描器的精度和常规的印象方法的准确性分析与vinylsiloxanether材料。 表1显示了正确度与表2中的主模型的精度从不同的xyz轴方向和从常规的印象法扫描。从不同方向扫描验证参考扫描器精度牙科精度这一特定需求。它揭示了扫描软件的任何滤镜效果。结果表明，在整个牙弓非常低偏差。的差值图像可以用于可视化的扫描过程的特定局限性。与陡峭的斜坡状的门齿的腭方面领域存在，因为较低的更高的局部偏差表面的扫描质量（ 图19）。 不同造成的印象的方法的偏差均小于基准扫描系统的内部精度显著更高。既正确度与精密度均显著低于比与参考扫描仪的传统印象的方法。的差值图像显示的印象的变形带有正负的变形特别是对牙弓（ 图20A）的前端。数字印象表现出不同种偏差图案，具有较高的偏差特别面向远端的齿（ 图20B） 牙科材料的磨损可以以相同的方式进行分析，前和咀嚼仿真后叠加在牙齿表面。 图21显示了前和后的咀嚼型模拟及其叠加试样。当地面亏损为六sible中的差分图像。高度的平均垂直损失和量的变化可以用差异分析软件进行测量。 <img alt="图1" fo:content-width="5in" src="/files/ftp_upload/51374/51374fig1highres.jpg" width="500" /> 图1:试样用平底放在参考扫描仪的XY工作台 <a href="https://www.jove.com/files/ftp_upload/51374/51374fig1highres.jpg" target="_blank">点击这里查看大图。</a> <img alt="图2" fo:content-width="5in" src="/files/ftp_upload/51374/51374fig2highres.jpg" width="500" /> 图2:启动"中频实验室测量模块。" <a href="https://www.jove.com/files/ftp_upload/51374/51374fig2highres.jpg" target="_blank">点击此处查看大图。</a> 图3:调整z轴的基地。 <a href="https://www.jove.com/files/ftp_upload/51374/51374fig3highres.jpg" target="_blank">点击这里查看大图。</a> <img alt="图4" fo:content-width="5in" src="/files/ftp_upload/51374/51374fig4highres.jpg" width="500" /> 图4:"对比度"调整"曝光"，并 <a href="https://www.jove.com/files/ftp_upload/51374/51374fig4highres.jpg" target="_blank">点击这里查看大图。</a> <img alt="图5" fo:content-width="5in" src="/files/ftp_upload/51374/51374fig5highres.jpg" width="500" /> 图5:定义扫描参数。 </st<html>荣&gt; <a href="https://www.jove.com/files/ftp_upload/51374/51374fig5highres.jpg" target="_blank">点击这里查看大图。</a> <img alt="图6" fo:content-width="5in" src="/files/ftp_upload/51374/51374fig6highres.jpg" width="500" /> 图6:定义扫描体积。 <a href="https://www.jove.com/files/ftp_upload/51374/51374fig6highres.jpg" target="_blank">点击这里查看大图。</a> <img alt="图7" fo:content-width="5in" src="/files/ftp_upload/51374/51374fig7highres.jpg" width="500" /> 图7:抽取点的大小。 <a href="https://www.jove.com/files/ftp_upload/51374/51374fig7highres.jpg" target="_blank">点击这里查看大图。</a> s/ftp_upload/51374/51374fig8highres.jpg"宽度="500"/&gt; 图8:试样的预扫 <a href="https://www.jove.com/files/ftp_upload/51374/51374fig8highres.jpg" target="_blank">点击这里查看大图。</a> <img alt="图9" fo:content-width="5in" src="/files/ftp_upload/51374/51374fig9highres.jpg" width="500" /> 图9:定义需要的图像信息。 <a href="https://www.jove.com/files/ftp_upload/51374/51374fig9highres.jpg" target="_blank">点击这里查看大图。</a> <img alt="图10" fo:content-width="5in" src="/files/ftp_upload/51374/51374fig10highres.jpg" width="500" /> 图10:控制扫描 <a href="https://www.jove.com/files/ftp_upload/51374/51374fig10highres.jpg" target="_blank">点击这里查看大图。</a> <img alt="图11" fo:content-width="5in" src="/files/ftp_upload/51374/51374fig11highres.jpg" width="500" /> 图11:扫描控制重复性 <a href="https://www.jove.com/files/ftp_upload/51374/51374fig11highres.jpg" target="_blank">点击这里查看大图。</a> <img alt="图12" fo:content-width="5in" src="/files/ftp_upload/51374/51374fig12highres.jpg" width="500" /> 图12:加载第二个模型扫描的差异分析。 <a href="https://www.jove.com/files/ftp_upload/51374/51374fig12highres.jpg" target="_blank">点击这里查看大图。</a> <img alt="图13" fo:content-width="5in" src="/files/ftp_upload/51374/51374fig13highres.jpg" width="500" /> FIGURË13:前自动最佳报名两种扫描数据集手动调整 <a href="https://www.jove.com/files/ftp_upload/51374/51374fig13highres.jpg" target="_blank">点击这里查看大图。</a> <img alt="图14" fo:content-width="5in" src="/files/ftp_upload/51374/51374fig14highres.jpg" width="500" /> 图14:设置刻度的视觉差异分析 <a href="https://www.jove.com/files/ftp_upload/51374/51374fig14highres.jpg" target="_blank">点击这里查看大图。</a> <img alt="图15" fo:content-width="5in" src="/files/ftp_upload/51374/51374fig15highres.jpg" width="500" /> 图15:显示和出口统计数据文件进行分析 <a href="https://www.jove.com/files/ftp_upload/51374/51374fig15highres.jpg" target="_blank">点击这里</a><html>查看大图。 <img alt="图16" fo:content-width="5in" src="/files/ftp_upload/51374/51374fig16highres.jpg" width="500" /> 图16:对重复性的扫描参数的影响 A）扫描单齿模型磨损测量。用于扫描新材料的第一步是找到最佳的对比度和亮度，以确保高重复性的精确测量B）一个风口浪尖上的重复性不理想扫描参数（对比度:0.25，亮度:0.8毫秒），C）模型的重复性表面具有最佳的扫描参数（对比度1.3，亮度1.4毫秒）。 <a href="https://www.jove.com/files/ftp_upload/51374/51374fig16highres.jpg" target="_blank">点击这里查看大图。</a> jpg"格式宽度="500"/&gt; 图17:准备一个完整的拱模型扫描一）无基B）重复性的模式利润扫描是因为牙齿表面的高角度低，尤其是前牙区域C）的不同扫描方向上的叠加显示扫描变形，开始在模型D的前部区域）周围的模型具有相同的光反射特性的材料E）扫描模式和基地展示高重复性各地的扫描对象F）不同的叠加应用水平底座扫描方向只显示了小的局部偏差，但扫描没有整个变形。 <a href="https://www.jove.com/files/ftp_upload/51374/51374fig17highres.jpg" target="_blank">点击这里查看大图。</a> <img alt="图18" fo:content-width="5in" src="/files/ftp_upload/51374/51374fig18highres.jpg" width="500" /><br/&gt; 图18:协议对扫描仪的评估和测试精度为传统的全拱印象 <a href="https://www.jove.com/files/ftp_upload/51374/51374fig18highres.jpg" target="_blank">点此查看大图。</a> <img alt="图19" fo:content-width="5in" src="/files/ftp_upload/51374/51374fig19highres.jpg" width="500" /> 图19:差分分析软件，计算在两个表面之间的差值与从每一个表面点的签名近邻法，致使每个比较周围6000000差值。所不同的数据可以导出为统计分析的Excel文件。 <a href="https://www.jove.com/files/ftp_upload/51374/51374fig19highres.jpg" target="_blank">点击这里查看大图。</a> <im克ALT ="图20"FO:内容宽度="5英寸"SRC ="/ files/ftp_upload/51374/51374fig20highres.jpg"宽度="500"/&gt; 图20:不同的印象，方法差异的图像与主模型 （真实度） 相比 ，颜色从-100微米（紫色）渐变至+100微米（橙色）。的差分图像显示不同的偏差而不同印模材料（A和B）以及使用的数字印象方法（C）的模式。三维评价可以用来优化印象程序。 <a href="https://www.jove.com/files/ftp_upload/51374/51374fig20highres.jpg" target="_blank">点击这里查看大图。</a> <img alt="图21" fo:content-width="5in" src="/files/ftp_upload/51374/51374fig21highres.jpg" width="500" /> 图21:前后咀嚼模拟 ，颜色GR 后的试样之间的差分图像从-20微米（橙色）aded至-500微米（紫色）。牙齿表面的高分辨率扫描是用来评价不同修复材料的磨损，在体内和体外试验。 <a href="https://www.jove.com/files/ftp_upload/51374/51374fig21highres.jpg" target="_blank">点击这里查看大图。</a> <table border="1" cellpadding="0" cellspacing="0"><tbody><tr><td></td><td>扫描1 </td><td>扫描2 </td><td>扫描3 </td><td>扫描4 </td><td> 5扫描</td><td>平均值±标准差</td></tr><tr><td>参考扫描器</td><td> 5.5 </td><td> 6.5 </td><td> 5.0 </td><td> 6 </td><td> 5.5 </td><td> 5.3±1.1 </td></tr><tr><td>传统印象</td><td> 15.5 </td><td> 22.0 </td><td> 22.5 </td><td> 18.0 </td><td> 21.5 </td><td> 20.4±2.2 </td></tr></tbody></table> 表1:正确度 （[90-10] / 2％百分位数，平均参考扫描仪和传统印象vinylsiloxanether的±标准偏差，微米）。 <table border="1" cellpadding="0" cellspacing="0"><tbody><tr><td></td><td> 1 </td><td> 2 </td><td> 3 </td><td> 4 </td><td> 5 </td><td> 6 </td><td> 7 </td><td> 8 </td><td> 9 </td><td> 10 </td><td></td></tr><tr><td>参考扫描器</td><td> 0.5 </td><td> 1.0 </td><td> 1.0 </td><td> 2.0 </td><td> 2.5 </td><td> 2.0 </td><td> 2.0 </td><td> 1.5 </td><td> 2.0 </td><td> 1.0 </td><td> 1.9±1.3 </td></tr><tr><td>传统印象</td><td> 10.5 </td><td> 11.0 </td><td> 11.0 </td><td> 14.5 </td><td> 14.5 </td><td> 16.5 </td><td> 10.0 </td><td> 11.0 </td><td> 15.5 </td><td> 10.5 </td><td> 12.5±2.5 </td></tr></tbody></table> 表2:精密 （[90-10] / 2个百分点，平均值±标准偏差振动性，参照扫描仪和常规的印象vinylsiloxanether的微米）。

Watch this Scientific Journal Video about Accuracy in Dental Medicine, A New Way to Measure Trueness and Precision at JoVE.com

Accuracy in Dental Medicine, A New Way to Measure Trueness and Precision

精度是口腔医学的一大需求。要验证的准确性，需要参考的扫描仪。本文提出了一种新的参考扫描仪获取各种各样的牙齿形态与高正确度与精密度的调整扫描方法。

精度在牙科医学，一种新的方法来测量正确度与精密度

Reference scanners are used in dental medicine to verify a lot of procedures. The main interest is to verify impression methods as they serve as a base ...

accuracy-dental-medicine-new-way-to-measure-trueness

Nanyang Technological University

Research

JoVE Journal

Medicine

13.3K 次观看.  University of Zürich. 精度是口腔医学的一大需求。要验证的准确性，需要参考的扫描仪。本文提出了一种新的参考扫描仪获取各种各样的牙齿形态与高正确度与精密度的调整扫描方法。 

视频: Accuracy in Dental Medicine, A New Way to Measure Trueness and Precision

Technique to Collect Fungiform (Taste) Papillae from Human Tongue

The sense of taste is critical for human life. It informs the body about the quality of food that will be potentially ingested and stimulates metabolic processes that prepare the alimentary canal for digestion. Steady progress is being made towards understanding the early biochemical and molecular events underlying taste transduction (for a review, Breslin and Spector, 20081). However, progress to date has largely resulted from animal models. Yet, since marked differences in receptor specificity and receptor density vary among species, human taste transduction will only be understood by using human taste tissue. Here we describe a biopsy technique to collect human fungiform papillae, visible as rounded pink anterior structures, about 0.5 mm in diameter that contain taste buds. These biopsied papillae are used for several purposes including the isolation of viable taste bud cells, in situ hybridization, immunohistochemistry and, through techniques of molecular biology, the identification of taste-specific novel proteins.

Technique to Collect Fungiform Taste Papillae from Human Tongue

Knowledge of molecular mechanisms underlying gustatory transduction has recently enjoyed significant advances, largely due to using animal models. However, the wide diversity in taste sensitivity and specificity among mammals warrants studies in human tissue. We describe a biopsy technique to collect living taste cells from the papillae on human tongue.

技术收集人舌菌状（味道）乳头

The sense of taste is critical for human life. It informs the body about the quality of food that will be potentially ingested and stimulates metabolic ...

科研

医学

Rapid Point-of-Care Assay of Enoxaparin Anticoagulant Efficacy in Whole Blood

There is the need for a clinical assay to determine the extent to which a patient's blood is effectively anticoagulated by the low-molecular-weight-heparin (LMWH), enoxaparin. There are also urgent clinical situations where it would be important if this could be determined rapidly. The present assay is designed to accomplish this. We only assayed human blood samples that were spiked with known concentrations of enoxaparin. The essential feature of the present assay is the quantification of the efficacy of enoxaparin in a patient's blood sample by degrading it to complete inactivity with heparinase. Two blood samples were drawn into Vacutainer tubes (Becton-Dickenson; Franklin Lakes, NJ) that were spiked with enoxaparin; one sample was digested with heparinase for 5 min at 37 &deg;C, the other sample represented the patient's baseline anticoagulated status. The percent shortening of clotting time in the heparinase-treated sample, as compared to the baseline state, yielded the anticoagulant contribution of enoxaparin. We used the portable, battery operated Hemochron 801 apparatus for measurements of clotting times (International Technidyne Corp., Edison, NJ). The apparatus has 2 thermostatically controlled (37 &deg;C) assay tube wells. We conducted the assays in two types of assay cartridges that are available from the manufacturer of the instrument. One cartridge was modified to increase its sensitivity. We removed the kaolin from the FTK-ACT cartridge by extensive rinsing with distilled water, leaving only the glass surface of the tube, and perhaps the detection magnet, as activators. We called this our minimally activated assay (MAA). The use of a minimally activated assay has been studied by us and others. 2-4 The second cartridge that was studied was an activated partial thromboplastin time (aPTT) assay (A104). This was used as supplied from the manufacturer. The thermostated wells of the instrument were used for both the heparinase digestion and coagulation assays. The assay can be completed within 10 min. The MAA assay showed robust changes in clotting time after heparinase digestion of enoxaparin over a typical clinical concentration range. At 0.2 anti-Xa I.U. of enoxaparin per ml of blood sample, heparinase digestion caused an average decrease of 9.8% (20.4 sec) in clotting time; at 1.0 I.U. per ml of enoxaparin there was a 41.4% decrease (148.8 sec). This report only presents the experimental application of the assay; its value in a clinical setting must still be established.

Demonstration of a rapid quantitative assay of the inhibition of blood coagulation by the low-molecular-weight-heparin, enoxaparin. The contribution of enoxaparin is assessed by removing its influence through digestion with heparinase. A fuller description of the assay is detailed in our published paper.1 The assay still requires clinical confirmation.

快速点护理依诺肝素的抗凝血剂功效的测定全血中

There is the need for a clinical assay to determine the extent to which a patient's blood is effectively anticoagulated by the ...

Construction of Vapor Chambers Used to Expose Mice to Alcohol During the Equivalent of all Three Trimesters of Human Development

Exposure to alcohol during development can result in a constellation of morphological and behavioral abnormalities that are collectively known as Fetal Alcohol Spectrum Disorders (FASDs). At the most severe end of the spectrum is Fetal Alcohol Syndrome (FAS), characterized by growth retardation, craniofacial dysmorphology, and neurobehavioral deficits. Studies with animal models, including rodents, have elucidated many molecular and cellular mechanisms involved in the pathophysiology of FASDs. Ethanol administration to pregnant rodents has been used to model human exposure during the first and second trimesters of pregnancy. Third trimester ethanol consumption in humans has been modeled using neonatal rodents. However, few rodent studies have characterized the effect of ethanol exposure during the equivalent to all three trimesters of human pregnancy, a pattern of exposure that is common in pregnant women. Here, we show how to build vapor chambers from readily obtainable materials that can each accommodate up to six standard mouse cages. We describe a vapor chamber paradigm that can be used to model exposure to ethanol, with minimal handling, during all three trimesters. Our studies demonstrate that pregnant dams developed significant metabolic tolerance to ethanol. However, neonatal mice did not develop metabolic tolerance and the number of fetuses, fetus weight, placenta weight, number of pups/litter, number of dead pups/litter, and pup weight were not significantly affected by ethanol exposure. An important advantage of this paradigm is its applicability to studies with genetically-modified mice. Additionally, this paradigm minimizes handling of animals, a major confound in fetal alcohol research.

We demonstrate the construction of alcohol vapor chambers using readily available materials that simultaneously house 6 mouse cages. We further describe their use in a mouse model of fetal alcohol exposure equivalent to all 3&#160;trimesters of human pregnancy. This paradigm exposes animals during gestation and postnatal days 1-12.

蒸汽腔体建设用于公开小鼠酒精在所有三个学期人类发展的等效

Exposure to alcohol during development can result in a constellation of morphological and behavioral abnormalities that are collectively known as Fetal ...

Intrauterine Telemetry to Measure Mouse Contractile Pressure In Vivo

A complex integration of molecular and electrical signals is needed to transform a quiescent uterus into a contractile organ at the end of pregnancy. Despite the discovery of key regulators of uterine contractility, this process is still not fully understood. Transgenic mice provide an ideal model in which to study parturition. Previously, the only method to study uterine contractility in the mouse was ex vivo isometric tension recordings, which are suboptimal for several reasons. The uterus must be removed from its physiological environment, a limited time course of investigation is possible, and the mice must be sacrificed. The recent development of radiometric telemetry has allowed for longitudinal, real-time measurements of in vivo intrauterine pressure in mice. Here, the implantation of an intrauterine telemeter to measure pressure changes in the mouse uterus from mid-pregnancy until delivery is described. By comparing differences in pressures between wild type and transgenic mice, the physiological impact of a gene of interest can be elucidated. This technique should expedite the development of therapeutics used to treat myometrial disorders during pregnancy, including preterm labor.

Intrauterine Telemetry to Measure Mouse Contractile Pressure In Vivo

This manuscript provides a protocol for implanting telemeters in the mouse for the purpose of measuring intrauterine pressures during pregnancy.

宫内遥测来衡量鼠标收缩压力<em&gt;在体内</em

A complex integration of molecular and electrical signals is needed to transform a quiescent uterus into a contractile organ at the end of pregnancy. ...

Studying Pancreatic Cancer Stem Cell Characteristics for Developing New Treatment Strategies

Pancreatic ductal adenocarcinoma (PDAC) contains a subset of exclusively tumorigenic cancer stem cells (CSCs) which have been shown to drive tumor initiation, metastasis and resistance to radio- and chemotherapy. Here we describe a specific methodology for culturing primary human pancreatic CSCs as tumor spheres in anchorage-independent conditions. Cells are grown in serum-free, non-adherent conditions in order to enrich for CSCs while their more differentiated progenies do not survive and proliferate during the initial phase following seeding of single cells. This assay can be used to estimate the percentage of CSCs present in a population of tumor cells. Both size (which can range from 35 to 250 micrometers) and number of tumor spheres formed represents CSC activity harbored in either bulk populations of cultured cancer cells or freshly harvested and digested tumors 1,2. Using this assay, we recently found that metformin selectively ablates pancreatic CSCs; a finding that was subsequently further corroborated by demonstrating diminished expression of pluripotency-associated genes/surface markers and reduced in vivo tumorigenicity of metformin-treated cells. As the final step for preclinical development we treated mice bearing established tumors with metformin and found significantly prolonged survival. Clinical studies testing the use of metformin in patients with PDAC are currently underway (e.g., NCT01210911, NCT01167738, and NCT01488552). Mechanistically, we found that metformin induces a fatal energy crisis in CSCs by enhancing reactive oxygen species (ROS) production and reducing mitochondrial transmembrane potential. In contrast, non-CSCs were not eliminated by metformin treatment, but rather underwent reversible cell cycle arrest. Therefore, our study serves as a successful example for the potential of in vitro sphere formation as a screening tool to identify compounds that potentially target CSCs, but this technique will require further in vitro and in vivo validation to eliminate false discoveries.

Pancreatic cancer stem cells (CSCs) can be expanded in vitro using the anchorage-independent sphere culture technique, which represents a powerful tool to study CSC biology and can serve as the first step to develop novel CSC-targeting therapies. Here the methodology for expanding, analyzing and targeting of pancreatic CSCs is provided.

研究胰腺癌干细胞特性开发新的治疗策略

Pancreatic ductal adenocarcinoma (PDAC) contains a subset of exclusively tumorigenic cancer stem cells (CSCs) which have been shown to drive tumor ...

A Simple Device to Rapidly Prepare Whole Mounts of the Mouse Intestine

Preparing whole mounts of the mouse small intestine and colon for subsequent analysis or quantification can be time consuming and difficult. We describe the use of a simple device to cut and &#8216;roll&#8217; mouse intestines to rapidly prepare whole mount preparations of superior and uniform quality to that which can be achieved by hand. The device comprises a base that holds 4 stainless steel rods and a top, which acts a cutting guide. The rods are inserted into the lumen of the small intestine [divided into thirds] and the colon. The rods and samples are then placed over a piece of filter paper or card into the holding slots in the base of the device. The top of the device is then positioned and serves as a cutting guide. The two angled sections in the center of the top piece are used to guide a knife or scalpel and cut the intestines longitudinally on the top of the rods. Once the intestinal sections have been cut, the top is removed and the card,&#160;tissue and rods gently removed from the device and placed on the bench. The rods are then gently rolled sideways to flatten and stick the intestinal segments onto the underlying piece of filter paper or card. The final preparation can then be examined or fixed and stored for later analysis. The preparations are invaluable for the study of intestinal changes in normal or genetically modified mouse models. The preparations have been used for the study and quantification of the effects of inflammation (colitis), damage, pre-cancerous lesions (aberrant crypt foci (ACFs) and mucin depleted foci (MDFs)) and polyps or tumors.

The use of a simple device to cut and &#8216;roll&#8217; mouse intestines to rapidly prepare whole mount preparations is described.

一个简单的设备，迅速准备小鼠肠的全坐骑

Preparing whole mounts of the mouse small intestine and colon for subsequent analysis or quantification can be time consuming and difficult. We describe ...

Boldness, Aggression, and Shoaling Assays for Zebrafish Behavioral Syndromes

A behavioral syndrome exists when specific behaviors interact under different contexts. Zebrafish have been test subjects in recent studies and it is important to standardize protocols to ensure proper analyses and interpretations. In our previous studies, we have measured boldness by monitoring a series of behaviors (time near surface, latency in transitions, number of transitions, and darts) in a 1.5 L trapezoidal tank. Likewise, we quantified aggression by observing bites, lateral displays, darts, and time near an inclined mirror in a rectangular 19 L tank. By dividing a 76 L tank into thirds, we also examined shoaling preferences. The shoaling assay is a highly customizable assay and can be tailored for specific hypotheses. However, protocols for this assay also must be standardized, yet flexible enough for customization. In previous studies, end chambers were either empty, contained 5 or 10 zebrafish, or 5 pearl danios (D. albolineatus). In the following manuscript, we present a detailed protocol and representative data that accompany successful applications of the protocol, which will allow for replication of behavioral syndrome experiments.

This manuscript describes the setup, implementation, and analysis of boldness, aggression, and shoaling in zebrafish and testing for the presence of a behavioral syndrome. A standardized approach for behavioral quantification will allow for easier comparison across studies. Modifications to this protocol are possible as each assay can be run individually.

气魄，侵略和浅水测定斑马鱼行为综合征

A behavioral syndrome exists when specific behaviors interact under different contexts. Zebrafish have been test subjects in recent studies and it is ...

Open Tracheostomy Gastric Acid Aspiration Murine Model of Acute Lung Injury Results in Maximal Acute Nonlethal Lung Injury

Acid pneumonitis is a major cause of sterile acute lung injury (ALI) in humans. Acid pneumonitis spans the clinical spectrum from asymptomatic to acute respiratory distress syndrome (ARDS), characterized by neutrophilic alveolitis, and injury to both alveolar epithelium and vascular endothelium. Clinically, ARDS is defined by acute onset of hypoxemia, bilateral patchy pulmonary infiltrates and non-cardiogenic pulmonary edema. Human studies have provided us with valuable information about the physiological and inflammatory changes in the lung caused by ARDS, which has led to various hypotheses about the underling mechanisms. Unfortunately, difficulties determining the etiology of ARDS, as well as a wide range of pathophysiology have resulted in a lack of critical information that could be useful in developing therapeutic strategies.
Translational animal models are valuable when their pathogenesis and pathophysiology accurately reproduce a concept proven in both in vitro and clinical settings. Although large animal models (e.g., sheep) share characteristics of the anatomy of human trachea-bronchial tree, murine models provide a host of other advantages including: low cost; short reproductive cycle lending itself to greater data acquisition; a well understood immunologic system; and a well characterized genome leading to the availability of a variety of gene deletion and transgenic strains. A robust model of low pH induced ARDS requires a murine ALI that targets mainly the alveolar epithelium, secondarily the vascular endothelium, as well as the small airways leading to the alveoli. Furthermore, a reproducible injury with wide differences between different injurious and non-injurious insults is important.
The murine gastric acid aspiration model presented here using hydrochloric acid employs an open tracheostomy and recreates a pathogenic scenario that reproduces the low pH pneumonitis injury in humans. Additionally, this model can be used to examine interaction of a low pH insult with other pulmonary injurious entities (e.g., food particles, pathogenic bacteria).

This protocol induces acute lung injury in a mouse that has close fidelity to the pathogenesis of acid pneumonitis observed in humans. We generate a maximal acute nonlethal low pH lung injury and account for differences in rodent-human anatomic respiratory structure using an open tracheostomy coupled with circumferential pressure release.

急性肺损伤开放气管切开胃酸吸入小鼠模型结果在急性最大非致命性肺损伤

Acid pneumonitis is a major cause of sterile acute lung injury (ALI) in humans. Acid pneumonitis spans the clinical spectrum from asymptomatic to acute ...

Oral Biofilm Sampling for Microbiome Analysis in Healthy Children

Oral biofilm and its molecular analysis provide a basis for investigating various dental research and clinical questions. Knowledge of biofilm composition leads to a better understanding of cariogenic and periopathogenic mechanisms. Microbial changes taking place in the oral cavity during childhood are of interest for several reasons. The evolution of the child oral microbiota and shifts in its composition need to be analyzed further to understand and possibly prevent the onset of disease. At the same time, advanced knowledge of the natural composition of oral biofilm is needed. Early stages of caries-free permanent dentition with healthy gums provide a widely unaffected subgingival habitat that can serve as an in situ baseline for studying features of oral health and disease. Analysis of children's oral biofilm during different stages in life is thus an important theme in the field. Modern molecular analysis methods can provide comprehensive information about the bacterial diversity of such biofilms. To enable microbiota data comparison, it is important to standardize each step in the procedure for molecular data generation. This procedure spans from clinical sampling, Next Generation Sequencing (NGS), bioinformatic data processing, to taxonomic interpretation. One of the most critical factors here is biofilm sampling. Sampling in children is even more challenging in particular due to limited space in subgingival areas. We thus focus on the use of paper points for subgingival sampling. This article provides a detailed protocol for oral biofilm sampling of the subgingival sulcus, the mucosa, and saliva in children.

Changes in the oral microbiome throughout childhood are of growing interest. Comparison of different microbiome studies reveals a lack of standardized sampling protocols. Limited space makes sampling the sound subgingival sulcus of children challenging. Paper point sampling is presented here in detail as the method of choice for this area.

口腔生物膜取样在健康儿童中的微生物分析

Oral biofilm and its molecular analysis provide a basis for investigating various dental research and clinical questions. Knowledge of biofilm composition ...

Conducting Maximal and Submaximal Endurance Exercise Testing to Measure Physiological and Biological Responses to Acute Exercise in Humans

Regular physical activity has a positive effect on human health, but the mechanisms controlling these effects remain unclear. The physiologic and biologic responses to acute exercise are predominantly influenced by the duration and intensity of the exercise regimen. As exercise is increasingly thought of as a therapeutic treatment and/or diagnostic tool, it is important that standardizable methodologies be utilized to understand the variability and to increase the reproducibility of exercise outputs and measurements of responses to such regimens. To that end, we describe two different cycling exercise regimens that yield different physiologic outputs. In a maximal exercise test, exercise intensity is continually increased with a greater workload resulting in an increasing cardiopulmonary and metabolic response (heart rate, stroke volume, ventilation, oxygen consumption and carbon dioxide production). In contrast, during endurance exercise tests, the demand is increased from that at rest, but is raised to a fixed submaximal exercise intensity resulting in a cardiopulmonary and metabolic response that typically plateaus. Along with the protocols, we provide suggestions on measuring physiologic outputs that include, but are not limited to, heart rate, slow and forced vital capacity, gas exchange metrics, and blood pressure to enable the comparison of exercise outputs between studies. Biospecimens can then be sampled to assess cellular, protein, and/or gene expression responses. Overall, this approach can be easily adapted into both short- and long-term effects of two distinct exercise regimens.

To assess the influence of exercise intensity on physiologic and biologic responses, two different exercise testing protocols were utilized. Methods outlining exercise testing on a cycle ergometer as an incremental maximal oxygen consumption test and endurance, steady state submaximal endurance test are described.

进行最大和次极大耐力运动测试, 测量人类急性运动的生理和生物学反应

Regular physical activity has a positive effect on human health, but the mechanisms controlling these effects remain unclear. The physiologic and biologic ...