Name: mechanical manipulation of neurons to control axonal development
Uploaded: 2011-04-10T14:00:00
Description: Watch this Scientific Journal Video about Mechanical Manipulation of Neurons to Control Axonal Development at JoVE.com

在这种方法的发展，我们非常感谢博士罗伯特E巴克斯鲍姆的重要贡献。

1。使玻璃针。 <ol><li>一个可调的微针车夫是用来制造一个约4毫米长，已关闭了坚实的梁锥尖的针。相对于长期的灵活的提示，这短短的4毫米的长度将限制在实验过程中针尖的振动。在4毫米的光纤近端地区，针锥迅速从玻璃管的直径在1毫米至15微米，而最远端1毫米的纤维直径为2.5微米。我们使用的R - 6玻璃毛细管，外径为0.9毫米，编号0.6毫米8“长度和BB - CH车夫。如果研究者感兴趣的是在测?...

<ol>
	<li>Zheng, J., Buxbaum, R. E., Heidemann, S. R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Measurements+of+growth+cone+adhesion+to+culture+surfaces+by+micromanipulation.">Measurements of growth cone adhesion to culture surfaces by micromanipulation.</a> J Cell Biol. 127, 2049-2060 (1994).</li><li>Chada, S., Lamoureux, P., Buxbaum, R. E., Heidemann, S. R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Cytomechanics+of+neurite+outgrowth+from+chick+brain+neurons.">Cytomechanics of neurite outgrowth from chick brain neurons.</a> J Cell Sci. 110, 1179-1186 (1997).</li><li>Heidemann, S. R., Lamoureux, P., Buxbaum, R. E., Haynes, L. W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> The Neuron in Tissue Culture. , 105-119 (1999).</li><li>Lamoureux, P., Altun-Gultekin, Z. F., Lin, C., Wagner, J. A., Heidemann, S. R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Rac+is+required+for+growth+cone+function+but+not+neurite+assembly.">Rac is required for growth cone function but not neurite assembly.</a> J Cell Sci. 110, 635-641 (1997).</li><li>Lamoureux, P., Ruthel, G., Buxbaum, R. E., Heidemann, S. R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Mechanical+tension+can+specify+axonal+fate+in+hippocampal+neurons.">Mechanical tension can specify axonal fate in hippocampal neurons.</a> J Cell Biol. 159, 499-508 (2002).</li><li>Lamoureux, P., Heidemann, S. R., Martzke, N. R., Miller, K. E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Growth+and+elongation+within+and+along+the+axon.">Growth and elongation within and along the axon.</a> Dev Neurobiol. 70, 135-149 (2010).</li><li>Dennerll, T. J., Lamoureux, P., Buxbaum, R. E., Heidemann, S. R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+cytomechanics+of+axonal+elongation+and+retraction.">The cytomechanics of axonal elongation and retraction.</a> J Cell Biol. 109, 3073-3083 (1989).</li><li>Heidemann, S. R., Kaech, S., Buxbaum, R. E., Matus, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Direct+observations+of+the+mechanical+behaviors+of+the+cytoskeleton+in+living+fibroblasts.">Direct observations of the mechanical behaviors of the cytoskeleton in living fibroblasts.</a> J Cell Biol. 145, 109-122 (1999).</li><li>Yoneda, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Force+Exerted+by+a+Single+Cilium+of+Mytilus-Edulis+.1.">Force Exerted by a Single Cilium of Mytilus-Edulis .1.</a> Journal of Experimental Biology. 37, (1960).</li><li>Bray, D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Mechanical+Tension+Produced+by+Nerve-Cells+in+Tissue-Culture.">Mechanical Tension Produced by Nerve-Cells in Tissue-Culture.</a> Journal of Cell Science. 37, 391-410 (1979).</li><li>Pfister, B. J., Iwata, A., Meaney, D. F., Smith, D. H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Extreme+stretch+growth+of+integrated+axons.">Extreme stretch growth of integrated axons.</a> J Neurosci. 24, 7978-7983 (2004).</li><li>Fass, J. N., Odde, D. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Tensile+force-dependent+neurite+elicitation+via+anti-beta1+integrin+antibody-coated+magnetic+beads.">Tensile force-dependent neurite elicitation via anti-beta1 integrin antibody-coated magnetic beads.</a> Biophys J. 85, 623-636 (2003).</li><li>Yang, S., Saif, M. T. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Microfabricated+Force+Sensors+and+Their+Applications+in+the+Study+of+Cell+Mechanical+Response.">Microfabricated Force Sensors and Their Applications in the Study of Cell Mechanical Response.</a> Exp Mech. 49, 135-151 (2009).</li><li>Bernal, R., Melo, F., Pullarkat, P. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Drag+Force+as+a+Tool+to+Test+the+Active+Mechanical+Response+of+PC12+Neurites.">Drag Force as a Tool to Test the Active Mechanical Response of PC12 Neurites.</a> Biophysical Journal. 98, 515-523 (2010).</li><li>Lamoureux, P., Buxbaum, R. E., Heidemann, S. R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Direct+evidence+that+growth+cones+pull.">Direct evidence that growth cones pull.</a> Nature. 340, 159-162 (1989).</li><li>Heidemann, S. R., Lamoureux, P., Buxbaum, R. E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Growth+cone+behavior+and+production+of+traction+force.">Growth cone behavior and production of traction force.</a> J Cell Biol. 111, 1949-1957 (1990).</li><li>O'Toole, M., Lamoureux, P., Miller, K. 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+physical+model+of+axonal+elongation:+force,+viscosity,+and+adhesions+govern+the+mode+of+outgrowth.">A physical model of axonal elongation: force, viscosity, and adhesions govern the mode of outgrowth.</a> Biophys J. 94, 2610-2620 (2008).</li><li>Bray, D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Axonal+growth+in+response+to+experimentally+applied+mechanical+tension.">Axonal growth in response to experimentally applied mechanical tension.</a> Dev Biol. 102, 379-389 (1984).</li><li>Heidemann, S. R., Lamoureux, P., Ngo, K., Reynolds, M., Buxbaum, R. E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Open-dish+incubator+for+live+cell+imaging+with+an+inverted+microscope.">Open-dish incubator for live cell imaging with an inverted microscope.</a> Biotechniques. 35, 708-708 (2003).</li></ol>

技术应用和测量细胞的力量，有着悠久的历史9。我们的方法最初是由丹尼斯布雷，谁使用玻璃针，类似我们的“拖”的神经元在一个恒定的速率使用机动液压设备10的工作积极性。有许多细胞，其中包括应用力量的替代手段：步进电机11，磁珠12日，13微束和流体流动14。后者是类似于我们的做法，在蜂窝探头最终是对一个已知的刚度弯曲梁校准和蜂窝测量?...

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		<div>
		<table border="1">
		<thead>
		<tr>
		<th>Material Name</th>
		<th>Type</th>
		<th>Company</th>
		<th>Catalogue Number</th>
		<th>Comment</th>
		</tr>
		</thead>
		<tbody>
		
<tr>
<td>R-6 cap. Tube</td>
<td>&nbsp;</td>
<td>Drummond Scientific Co., Broomall, PA, USA</td>
<td>9-000-3111</td>
<td>R-6 glass OD 0.9mm, ID 0.6 mm, 8"</td>
</tr>
<tr>
<td>BB-CH puller</td>
<td>&nbsp;</td>
<td>Mecanex S.A., Geneva, Switzerland</td>
<td>BB-CH puller</td>
<td>Use Mode 4 Alt by CP=100, PP=10, SP1=1000, SP2=1000</td>
</tr>
<tr>
<td>0.001" Chromel wire</td>
<td>&nbsp;</td>
<td>Omega Engineering, Stamford, CT, USA</td>
<td>SPCH-001-50</td>
<td>unsheathed, themocouple wire, 50ft spool now called Chromega</td>
</tr>
<tr>
<td>0.003" Constatan wire</td>
<td>&nbsp;</td>
<td>Omega Engineering, Stamford, CT, USA</td>
<td>SPCI-003-50</td>
<td>unsheathed, themocouple wire, 50 ft spool</td>
</tr>
<tr>
<td>fine forceps</td>
<td>&nbsp;</td>
<td>Fine Science Tools, USA</td>
<td>91150-20</td>
<td>Dumont Inox #5</td>
</tr>
<tr>
<td>universal microscope boom stand</td>
<td>&nbsp;</td>
<td>Nikon</td>
<td>76135 or 90430</td>
<td>most brands or types of boom stand will work for this use</td>
</tr>
<tr>
<td>mechanical micromanipulator</td>
<td>&nbsp;</td>
<td>Narishige </td>
<td>M-152</td>
<td>three-axis direct-drive coarse micromanipulator</td>
</tr>
<tr>
<td>hydraulic micromanipulator</td>
<td>&nbsp;</td>
<td>Narishige </td>
<td>MO-203</td>
<td>now available as MMO-203, three movable axis type</td>
</tr>
<tr>
<td>needle holder</td>
<td>&nbsp;</td>
<td>Leica Microsystems</td>
<td>11520145</td>
<td>set of 3</td>
</tr>
<tr>
<td>single instrument holder</td>
<td>&nbsp;</td>
<td>Leica Microsystems</td>
<td>11520142</td>
<td>&nbsp;</td>
<tr>
<td>double instrument holder</td>
<td>&nbsp;</td>
<td>Leica Microsystems</td>
<td>11520143</td>
<td>&nbsp;</td>
<tr>
<td>mechanical micromanipulator</td>
<td>&nbsp;</td>
<td>Leica Microsystems</td>
<td>39430001</td>
<td>post mount,1 prob holder, RH Model 430001</td>
</tr>
<tr>
<td>joystick mech. micromanipulator</td>
<td>&nbsp;</td>
<td>Leica Microsystems</td>
<td>11520137</td>
<td>&nbsp;</td>
<tr>
<td>Leica DM IRB</td>
<td>&nbsp;</td>
<td>Leica Microsystems</td>
<td>&nbsp;</td>
<td>inverted microscope</td>
</tr>
<tr>
<td>Vibraplane isolation table</td>
<td>&nbsp;</td>
<td>Kinetic System, Boston, MA, USA</td>
<td>1200 series</td>
<td>ours is model 1201-02-12</td>
</tr>
<tr>
<td>Ringcubator</td>
<td>&nbsp;</td>
<td>self manufactured see reference 19</td>
<td>&nbsp;</td>
<td>reference 19, requires updated controller listed below</td>
</tr>
<tr>
<td>programable temperature controller</td>
<td>&nbsp;</td>
<td>Instrumart.com</td>
<td>Fuji Electric PXR3</td>
<td>replaces the retired PXV3 temperature controller</td>
</tr>
<tr>
<td>Nikon Diaphot TMD</td>
<td>&nbsp;</td>
<td>Nikon Instruments, Inc.</td>
<td>&nbsp;</td>
<td>inverted microscope, circa 1980</td>
</tr>
<tr>
<td>Nikon SMZ-10 binocular dissecting </td>
<td>&nbsp;</td>
<td>Nikon Instruments, Inc.</td>
<td>&nbsp;</td>
<td>other dissecting microscopes will work</td>
</tr>		</tbody>
		</table>
		</div>

mechanical manipulation of neurons to control axonal development

细胞操作和神经轴突的延伸，可以校准玻璃微纤维能够测量和应用在10-1000μdyne范围1,2的力量来完成。力的测量是通过观察Hookean弯曲的玻璃针，这是直接和实证方法3校准获得。设备制造，校准，处理和使用的针头对细胞的要求和程序进行了全面介绍。力制度与以前使用的不同的细胞类型，这些技术已应用示范的方法的灵活性，并为今后的调查4-6的例子给出。技术优势是连续的“可视化”的操作所产生的力量和能力在各种细胞活动的直接干预。这些措施包括直接刺激和调节轴突生长和回缩 7，以及分队，对任何类型的培养细胞 8机械测量。

Watch this Scientific Journal Video about Mechanical Manipulation of Neurons to Control Axonal Development at JoVE.com

Mechanical Manipulation of Neurons to Control Axonal Development

应用在2-1000 microdyne范围内的神经元上的力量和直接测量是实现与高精密校准的玻璃针。这种方法可以用来控制和测量轴突发展的几个方面，包括轴突起始，轴突紧张，轴突伸长速度，和力矢量。

机械操纵控制神经元轴突的发展

Cell manipulations and extension of neuronal axons can be accomplished with calibrated glass micro-fibers capable of measuring and applying forces in the ...

Research

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Dissection and Culture of Commissural Neurons from Embryonic Spinal Cord

从胚胎脊髓​​连合神经的解剖和文化

Commissural neurons have been widely used to investigate the mechanisms underlying axon guidance during embryonic spinal cord development. The cell bodies ...

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Lentivirus-mediated Genetic Manipulation and Visualization of Olfactory Sensory Neurons in vivo

Lentivirus-mediated Genetic Manipulation and Visualization of Olfactory Sensory Neurons in vivo

嗅觉感官神经元的慢病毒介导的遗传操作和可视化在体内</em

Development of a precise olfactory circuit relies on accurate projection of olfactory sensory neuron (OSN) axons to their synaptic 
targets in the ...

DiI-Labeling of DRG Neurons to Study Axonal Branching in a Whole Mount Preparation of Mouse Embryonic Spinal Cord

DII的标签DRG神经元的研究轴突分支在小鼠胚胎脊髓整装准备

Here we  present a technique to label the trajectories of small groups of DRG neurons  into the embryonic spinal cord by diffusive staining using the ...

Single-cell Profiling of Developing and Mature Retinal Neurons

视网膜神经细胞发育和成熟的单细胞的剖析

Highly specialized, but exceedingly small populations of cells play important roles in many tissues. The identification of cell-type specific markers and ...

Odorant-induced Responses Recorded from Olfactory Receptor Neurons using the Suction Pipette Technique

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Animals sample the odorous environment around them through the chemosensory systems located in the nasal cavity. Chemosensory signals affect complex ...

A Method for High Fidelity Optogenetic Control of Individual Pyramidal Neurons In vivo

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单个锥体神经元的高保真光遗传学控制方法<em&gt;在体内</em

Optogenetic methods  have emerged as a powerful tool for elucidating neural circuit activity  underlying a diverse set of behaviors across a broad range ...

Genetic Manipulation of Cerebellar Granule Neurons In Vitro and In Vivo to Study Neuronal Morphology and Migration

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Developmental events in the brain including neuronal morphogenesis and migration are highly orchestrated processes. In vitro and in vivo analyses allow ...

Real-time Imaging of Axonal Transport of Quantum Dot-labeled BDNF in Primary Neurons

初级量子神经元轴突运输的实时成像点标记的BDNF

BDNF plays an important role in several facets of neuronal survival, differentiation, and function. Structural and functional deficits in axons are ...

Optical Control of a Neuronal Protein Using a Genetically Encoded Unnatural Amino Acid in Neurons

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Photostimulation is a noninvasive way to control biological events with excellent spatial and temporal resolution. New methods are desired to ...

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The rise of radio-enabled digital electronic devices has prompted the use of small wireless neuromuscular recorders and stimulators for studying in-flight ...