Name: dissection, microct scanning and morphometric analyses of the baculum
Uploaded: 2017-03-19T15:00:00
Description: Watch this Scientific Journal Video about Dissection, MicroCT Scanning and Morphometric Analyses of the Baculum at JoVE.com

蒂姆·戴利和安德鲁·史密斯在提供早期的许多有用的计算讨论;蒂姆·戴利写了协议5.计算资源是由高性能计算集群在南加州大学提供了必要的程序rotate_translate_cylindrical。这项工作是由美国国立卫生研究院授予＃GM098536（MDD）的支持。

所有的过程和人员由南加州学院的大学动物护理和使用委员会（IACUC），协议＃11394批准。 1.解剖短肛和准备<ol><li>通过二氧化碳过度曝光安乐死一个性成熟的雄性小鼠，根据相关机构的动物护理和使用委员会（IACUC）提出的协议。 </li><li>躺在仰卧位的动物，并通过施加压力拖延阴茎大拇指外侧包皮开幕。 </li><li>一旦阴茎旷日持久，通过包皮尽可能延长组织。 </li><li>用剪刀?...

<ol>
	<li>Slice, D. E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Geometrics+morphometrics.">Geometrics morphometrics.</a> Annu. Rev. Anthropol. 36, 261-281 (2007).</li><li>Slice, D. E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> Modern morphometrics in physical anthropology. 6, (2005).</li><li>Zelditch, M. L., Swiderski, D. L., Sheets, H. D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> Geometric morphometrics for biologists: a primer. , (2012).</li><li>Bookstein, 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> Morphometric tools for landmark data: geometry and biology. , (1991).</li><li>Rohlf, F. J., Marcus, L. 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+Revolution+in+Morphometrics.">A Revolution in Morphometrics.</a> Trends. Ecol. Evol. 8 (4), 129-132 (1993).</li><li>Zelditch, M. L., Swiderski, D. L., Sheets, H. D., Fink, W. 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> Geometric morphometrics for biologists: a primer. , (2004).</li><li>Rohlf, F. J., Slice, D. E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Extensions+of+the+Procrustes+method+for+the+optimal+superimposition+of+landmarks.">Extensions of the Procrustes method for the optimal superimposition of landmarks.</a> Syst. Zool. 39 (1), 40-59 (1990).</li><li>Gunz, P., Mitteroecker, P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Semilandmarks:+a+method+for+quantifying+curves+and+surfaces.">Semilandmarks: a method for quantifying curves and surfaces.</a> Hystrix. 24 (1), 103-109 (2013).</li><li>Gunz, P., Ramsier, M., Kuhrig, M., Hublin, J. J., Spoor, F. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+mammalian+bony+labyrinth+reconsidered,+introducing+a+comprehensive+geometric+morphometric+approach.">The mammalian bony labyrinth reconsidered, introducing a comprehensive geometric morphometric approach.</a> J. Anat. 220 (6), 529-543 (2012).</li><li>Mitteroecker, P., Gunz, P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Advances+in+geometric+morphometrics.">Advances in geometric morphometrics.</a> Evol. Biol. 36 (2), 235-247 (2009).</li><li>Bookstein, F. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Landmark+methods+for+forms+without+landmarks:+morphometrics+of+group+differences+in+outline+shape.">Landmark methods for forms without landmarks: morphometrics of group differences in outline shape.</a> Med. Im. Anal. 1 (3), 225-243 (1997).</li><li>Gunz, P., Mitteroecker, P., Bookstein, F., Slice, D. E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> Modern morphometrics in physical anthropology. , 73-98 (2005).</li><li>Oxnard, C., O'Higgins, P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Biology+Clearly+Needs+Morphometrics.+Does+Morphometrics+Need+Biology?.">Biology Clearly Needs Morphometrics. Does Morphometrics Need Biology?.</a> Biological Theory. 4 (1), 84-97 (2009).</li><li>Schultz, N. G., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+genetic+basis+of+baculum+size+and+shape+variation+in+mice.">The genetic basis of baculum size and shape variation in mice.</a> G3. 6 (5), 1141-1151 (2016).</li><li>Schultz, N. G., Lough-Stevens, M., Abreu, E., Orr, T. J., Dean, 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=The+baculum+was+gained+and+lost+multiple+times+during+mammalian+evolution.">The baculum was gained and lost multiple times during mammalian evolution.</a> Integr Comp Biol. 56 (4), 644-656 (2016).</li><li>Dines, J. P., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Sexual+selection+targets+cetacean+pelvic+bones.">Sexual selection targets cetacean pelvic bones.</a> Evolution. 68 (11), 3296-3306 (2014).</li><li>Adams, D. C., Ot&#225;rola-Castillo, E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=geomorph:+an+R+package+for+the+collection+and+analysis+of+geometric+morphometric+shape+data.">geomorph: an R package for the collection and analysis of geometric morphometric shape data.</a> Methods Ecol. Evol. 4 (4), 393-399 (2013).</li></ol>

在上述协议中的关键步骤是：1）解剖的bacula，2）收集显微图像，3）显微输出转换到XYZ的平面文件坐标，4）分割出各试样的点云，5）将每个试样标准化的坐标系，和6）限定的半标。这些步骤很容易被修改，以适应不同的对象。 这些方法可能可以被应用到实质上是“棒状”，或者至少不太弯曲的任何对象。对象曲线回自己成为“U形”不能被目前分析，因为切片（ 图1G?...

形态测量的领域包括的方法来量化的大小和生物形式的形状，在科学调查1，2，3，4，5，6的一个基本步骤的分集。传统上，大小和形状的统计分析开始于识别上的生物结构的地标，然后测量线性距离，角度和比例，这可以在一个多变量的框架进行分析。里程碑式的基于几何形态特征是保留的地标空间位置，通过分析和可视化5保存从数据收集几何信息的方法。广义普鲁克分析（GPA）可以应用到消除位置，规模，和地标旋转变化，以产生是m标本之间的对准inimizes他们的平方差-剩下的就是形状相异7。 任何形态分析的一个重要概念是同源性，或一个可以可靠地识别代表该标本或结构之间对应生物学意义的和离散的特征的地标的想法。例如，人头骨有同源流程，椎间孔，缝合线，和管道，可以使形态学分析。不幸的是，对应的地标标识是在许多生物结构困难，特别是那些具有光滑表面或曲线8，9，10。 我们下面的办法使用计算几何这个问题。的一般工作流程，以产生可以被表示为点的云的物体的三维扫描，然后旋转和转换，该转换点云，使得所有specimens被定向一个共同的坐标系上。然后我们数学定义从物体的特定区域的半标。放在这些地区离散半地标生物任意11。开展GPA和随后的统计分析可以产生不良的文物8，12，因为随意放置地标可能不是生物同源。因此，我们允许这些半地标数学“滑动”。此过程减少结构之间的电势差。如别处认为这里使用的滑动算法是适当的量化缺乏类似解剖区域容易识别相应的地标3，6，8，10，11，12。这些方法各有其利mitations 13，但应该适应不同尺寸和形状的物体。 这里，我们说明这个方法是如何在近一个鼠标阴茎骨14的研究应用，在已经获得和哺乳动物进化15期间丢失多个独立倍阴茎骨。我们讨论解剖和制备的特定骨的，所述阴茎骨（协议1），生成显微图像（协议2），并且这些图像以一种格式，使所有下游计算几何变换（协议3和4）。完成这些步骤后，由〜100K XYZ坐标每个试样表示。然后，我们通过一系列的变革，有效地调整所有标本到一个共同的方向（协议5）走，然后定义从对准标本半地标（协议6）。协议1-4应该是相似的，无论被分析的对象。协议5和6号议定书是SPEcifically专为阴茎骨，但我们希望通过详细介绍这些步骤，研究者可以想像的修改，这将是对有关他们感兴趣的对象。例如，这些方法改进了用于研究鲸骨盆和肋骨16。

<table border="0" cellpadding="0" cellspacing="0" width="743">
	<tbody>
		<tr height="42">
			<td height="42" style="height:42px;width:211px;">Dissecting scissors</td>
			<td style="width:287px;">VWR</td>
			<td style="width:79px;">470106-338</td>
			<td style="width:167px;">Most sizes should work</td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:211px;">Dissecting Forceps, Fine Tip, Curved</td>
			<td style="width:287px;">VWR</td>
			<td style="width:79px;">82027-406</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:211px;">1.7 mL microcentrifuge tube</td>
			<td style="width:287px;">VWR</td>
			<td style="width:79px;">87003-294</td>
			<td></td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:211px;">Absolute Ethanol</td>
			<td style="width:287px;">Fisher Scientific</td>
			<td style="width:79px;">CAS 64-17-5</td>
			<td>To be diluted to 70% for dissections</td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:211px;">Floral Foam</td>
			<td style="width:287px;">Wholesale Floral</td>
			<td style="width:79px;">6002-48-07</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">uCT50 scanner&#160;</td>
			<td colspan="2">Scanco Medical AG, Bruttisellen, Switzerland</td>
			<td></td>
		</tr>
	</tbody>
</table>

dissection, microct scanning and morphometric analyses of the baculum

现代形态测量提供了强大的方法来量化的大小和形状的变化。一个基本要求是，确定地标坐标列表;然而，这样的坐标必须代表整个标本同源结构。虽然许多生物对象包括容易识别标志，以满足同源性的假设，许多缺乏这种结构。一个可能的解决方案是在数学上横跨试样代表相同形态区域中的对象在地方半标。在这里，我们说明了一个新近开发的管道数学定义从鼠标阴茎骨（阴茎骨）半地标。我们的方法应该适用于广泛范围的对象。

在协议6中产生的半地标XYZ坐标可以直接导入到任何基于具有里程碑意义的几何形态测量分析17。计算管道上面已经用于研究小鼠的bacula 14，以及鲸骨盆和肋骨16。上半的地标计算定义的更多细节，这里提出，以试图帮助研究人员可视可能被修改，以适应他们感兴趣的特定对象的步骤。该阴茎骨包含了被利用来自动化某?...

Watch this Scientific Journal Video about Dissection, MicroCT Scanning and Morphometric Analyses of the Baculum at JoVE.com

Dissection, MicroCT Scanning and Morphometric Analyses of the Baculum

许多生物结构缺乏容易定义的地标，从而难以应用现代形态的方法。在这里，我们举例说明的方法来研究的小鼠阴茎骨（在阴茎骨），包括夹层和显微扫描，随后计算方法来定义用于量化的尺寸和形状的变化的半标。

在中短肛解剖，显微扫描和分析形态

Modern morphometrics provides powerful methods to quantify size and shape variation. A basic requirement is a list of coordinates that define landmarks; ...

Research

JoVE Journal

Bioengineering

Specimen Preparation, Imaging, and Analysis Protocols for Knife-edge Scanning Microscopy

刀口扫描显微镜的样品制备，图像和分析协议

Major advances in high-throughput, high-resolution, 3D microscopy techniques have enabled the acquisition of large volumes of neuroanatomical data at ...

科研

生物工程

Video-rate Scanning Confocal Microscopy and Microendoscopy

视频率扫描共聚焦显微镜和内窥镜

Confocal microscopy has become an invaluable tool in biology and the biomedical sciences, enabling rapid, high-sensitivity, and high-resolution optical ...

Evaluation of Biomaterials for Bladder Augmentation using Cystometric Analyses in Various Rodent Models

膀胱膀胱分析各种啮齿类动物模型中使用的增广的生物材料评价

Renal function and continence of urine are critically dependent on the proper function of the urinary bladder, which stores urine at low pressure and ...

Aplysia Ganglia Preparation for Electrophysiological and Molecular Analyses of Single Neurons

Aplysia Ganglia Preparation for Electrophysiological and Molecular Analyses of Single Neurons

阿普利西亚 单一神经元电生理学和分子分析的甘利亚制备

A major challenge in neurobiology is to understand the molecular underpinnings of neural circuitry that govern a specific behavior. Once the specific ...

Large-area Scanning Probe Nanolithography Facilitated by Automated Alignment and Its Application to Substrate Fabrication for Cell Culture Studies

基于自动对准的大面积扫描探针光刻及其在细胞培养研究中的应用

Scanning probe microscopy has enabled the creation of a variety of methods for the constructive (&#39;additive&#39;) top-down fabrication of ...

Automated Slide Scanning and Segmentation in Fluorescently-labeled Tissues Using a Widefield High-content Analysis System

用 Widefield 高含量分析系统实现荧光标记组织的自动滑动扫描和分割

Automated slide scanning and segmentation of fluorescently-labeled tissues is the most efficient way to analyze whole slides or large tissue sections. ...

Multimodal Volumetric Retinal Imaging by Oblique Scanning Laser Ophthalmoscopy (oSLO) and Optical Coherence Tomography (OCT)

Multimodal Volumetric Retinal Imaging by Oblique Scanning Laser Ophthalmoscopy oSLO and Optical Coherence Tomography OCT

斜扫描激光检眼镜 (奥斯陆) 与光学相干层析成像 (OCT) 的多模态视网膜成像

While fluorescence imaging is widely used in ophthalmology, a large field of view (FOV) three-dimensional (3D) fluorescence retinal image is still a big ...

Morphometric Protocol for the Objective Assessment of Blastocyst Behavior During Vitrification and Warming Steps

用于玻璃化和变暖步骤中囊胚行为客观评估的形态学协议

This article describes the noninvasive method of blastocyst morphometry based on time-lapse microphotography for the accurate monitoring of a ...

Evaluation of Photosynthetic Behaviors by Simultaneous Measurements of Leaf Reflectance and Chlorophyll Fluorescence Analyses

通过叶反射和叶绿素荧光分析同步测量的光合行为评价

Chlorophyll a fluorescence analysis is widely used to measure photosynthetic behaviors in intact plants, and has resulted in the development of many ...

Near Simultaneous Laser Scanning Confocal and Atomic Force Microscopy (Conpokal) on Live Cells

Near Simultaneous Laser Scanning Confocal and Atomic Force Microscopy Conpokal on Live Cells

近同步激光扫描在活细胞上的共和和原子力显微镜（康波卡尔）

Techniques available for micro- and nano-scale mechanical characterization have exploded in the last few decades. From further development of the scanning ...