Name: measurement of cellular chemotaxis with ecis/taxis
Uploaded: 2012-04-01T11:00:00
Description: Watch this Scientific Journal Video about Measurement of Cellular Chemotaxis with ECIS/Taxis at JoVE.com

这项工作是由来自美国国立卫生研究院（ES07408和EB00208）的赠款支持。

1。 ECIS的/出租车电极的制备<ol><li> ECIS的/的士电极的阵列（8％幻灯片商会组成）的金表面稳定在室温为15分钟，在无菌条件下用无菌去离子水（DH 2 O）的10毫米半胱氨酸的前处理。 </li><li>从每个电极室吸半胱氨酸溶液，冲洗与消毒卫生署2 O的 3倍，并更换完全培养基250μL（RPMI 1640培养液，10％胎牛血清，25毫米的HEPES缓冲液）。 </li><li>连接电极阵列联系仪器阵列支架引脚进行电极...

<ol>
	<li>Boyden, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+chemotactic+effect+of+mixtures+of+antibody+and+antigen+on+polymorphonuclear+leucocytes.">The chemotactic effect of mixtures of antibody and antigen on polymorphonuclear leucocytes.</a> J. Exp. Med. 115, 453-466 (1962).</li><li>Lauffenburger, D. A., Tranquillo, R. T., Zigmond, S. H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Concentration+gradients+of+chemotactic+factors+in+chemotaxis+assays.">Concentration gradients of chemotactic factors in chemotaxis assays.</a> Methods Enzymol. 162, 85-101 (1988).</li><li>Nelson, R. D., Quie, P. G., Simmons, R. L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Chemotaxis+under+agarose:+a+new+and+simple+method+for+measuring+chemotaxis+and+spontaneous+migration+of+human+polymorphonuclear+leukocytes+and+monocytes.">Chemotaxis under agarose: a new and simple method for measuring chemotaxis and spontaneous migration of human polymorphonuclear leukocytes and monocytes.</a> J. Immunol. 115, 1650-1650 (1975).</li><li>Newton-Nash, D. K., Tonellato, P., Swiersz, M., Abramoff, P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Assessment+of+chemokinetic+behavior+of+inflammatory+lung+macrophages+in+a+linear+under-agarose+assay.">Assessment of chemokinetic behavior of inflammatory lung macrophages in a linear under-agarose assay.</a> J. Leukoc. Biol. 48, 297-305 (1990).</li><li>Giaever, I., Keese, C. R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Monitoring+fibroblast+behavior+in+tissue+culture+with+an+applied+electric+field.">Monitoring fibroblast behavior in tissue culture with an applied electric field.</a> Proc. Natl. Acad. Sci. U.S.A. 81, 3761-374 (1984).</li><li>Keese, C. G. I. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+Whole+Cell+Biosensor+bsed+on+Cell-Substrate+Interactions.">A Whole Cell Biosensor bsed on Cell-Substrate Interactions.</a> IEEE Engineering in Medicine and Biology Society. 12, 500-501 (1990).</li><li>Laevsky, G., Knecht, D. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Cross-linking+of+actin+filaments+by+myosin+II+is+a+major+contributor+to+cortical+integrity+and+cell+motility+in+restrictive+environments.">Cross-linking of actin filaments by myosin II is a major contributor to cortical integrity and cell motility in restrictive environments.</a> J. Cell. Sci. 116, 3761-3770 (2003).</li><li>Condeelis, J., Bresnick, A., Demma, M., Dharmawardhane, S., Eddy, R., Hall, A. L., Sauterer, R., Warren, V. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Mechanisms+of+amoeboid+chemotaxis:+an+evaluation+of+the+cortical+expansion.">Mechanisms of amoeboid chemotaxis: an evaluation of the cortical expansion.</a> 11, 5-6 (1990).</li><li>Hadjout, N., Laevsky, G., Knecht, D. A., Lynes, M. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Automated+real-time+measurement+of+chemotactic+cell+motility.">Automated real-time measurement of chemotactic cell motility.</a> Biotechniques. 31, 1130-1138 (2001).</li><li>Hadjout, N., Yin, X., Knecht, D. A., Lynes, M. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Automated+real-time+measurements+of+leukocyte+chemotaxis.">Automated real-time measurements of leukocyte chemotaxis.</a> J. Immunol. Methods. 320, 70-80 (2007).</li><li>Yin, X., Knecht, D. A., Lynes, M. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Metallothionein+mediates+leukocyte+chemotaxis.">Metallothionein mediates leukocyte chemotaxis.</a> BMC Immunol. 6, 21-21 (2005).</li><li>Lundien, M. C., Mohammed, K. A., Nasreen, N., Tepper, R. S., Hardwick, J. A., Sanders, K. L., Van Horn, R. D., Antony, V. B. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Induction+of+MCP-1+expression+in+airway+epithelial+cells:+role+of+CCR2+receptor+in+airway+epithelial+injury.">Induction of MCP-1 expression in airway epithelial cells: role of CCR2 receptor in airway epithelial injury.</a> J. Clin. Immunol. 22, 144-152 (2002).</li><li>Zudaire, E., Cuesta, N., Murty, V. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+aryl+hydrocarbon+receptor+repressor+is+a+putative+tumor+suppressor+gene+in+multiple+human+cancers.">The aryl hydrocarbon receptor repressor is a putative tumor suppressor gene in multiple human cancers.</a> J. Clin. Invest. 118, 640-650 (2008).</li><li>Opp, D., Wafula, B., Lim, J., Huang, E., Lo, J. C., Lo, C. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Use+of+electric+cell-substrate+impedance+sensing+to+assess+in+vitro+cytotoxicity.">Use of electric cell-substrate impedance sensing to assess in vitro cytotoxicity.</a> Biosens. Bioelectron. 24, 2625-269 (2009).</li><li>Foxman, E. F., Kunkel, E. J., Butcher, E. C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Integrating+conflicting+chemotactic+signals.+The+role+of+memory+in+leukocyte+navigation.">Integrating conflicting chemotactic signals. The role of memory in leukocyte navigation.</a> J. Cell. Biol. 147, 577-588 (1999).</li><li>Heit, B., Tavener, S., Raharjo, E., Kubes, P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=An+intracellular+signaling+hierarchy+determines+direction+of+migration+in+opposing+chemotactic+gradients.">An intracellular signaling hierarchy determines direction of migration in opposing chemotactic gradients.</a> J. Cell. Biol. 159, 91-102 (1999).</li></ol>

ECIS的检测/出租车新颖等特点，包括其自动化实时数据的收集细胞以趋化因子的反应能力。虽然这项技术的最普遍的应用是测量细胞的反应，个别趋化梯度，或趋激动剂和拮抗剂的混合物组成的梯度，ECIS的/出租车方法的是服从这些配置的变化，这可能是非常有益的评估细胞的反应。有很好的证据，重叠或连续梯度可以影响细胞的行为，以新颖的方式。此外，它很可能是这些更为复杂的渐变是?...

<table border="1"><tbody><tr><td> 试剂名称 </td><td> 公司 </td><td> 目录编号 </td><td> 评论 </td></tr><tr><td> ECIS的Zθ </td><td>应用生物物理学</td><td></td><td> <a href="http://www.biophysics.com/prodducts_Ecisz0.php">http://www.biophysics.com/prodducts_Ecisz0.php</a> </td></tr><tr><td> ECIS的电极阵列</td><td>应用生物物理学</td><td> 8W趋</td><td> <a href="http://www.biophysics.com/cultureware.php">http://www.biophysics.com/cultureware.php</a> </td></tr><tr><td> sea​​kem GTG琼脂糖</td><td> BioWhittaker分子应用</td><td> 50070 </td><td></td></tr><tr><td>培养液</td><td> cellgro </td><td> 10-040 </td><td></td></tr><tr><td>胎牛血清胎牛血清白蛋白</td><td> Thermo Scientific的</td><td> SH300703 </td><td></td></tr><tr><td> PEnicillin /链霉素</td><td> MP的Biomedicals </td><td> 1670049 </td><td>青霉素5000国际单位/毫升，链霉素5毫克/毫升</td></tr><tr><td> HEPES缓冲</td><td> MP的Biomedicals </td><td> 1688449 </td><td> 1M的解决方案，细胞培养级</td></tr><tr><td> 14号不锈钢套管（2）4英寸</td><td>一般实验室供应</td><td> 5-8365-1 </td><td>钝点</td></tr></tbody></table>

measurement of cellular chemotaxis with ecis/taxis

在回应外界刺激细胞运动是许多细胞过程，包括伤口愈合，炎症和感染的反应的基础。一种常见的方法来衡量趋Boyden小室检测，其中由多孔膜分离细胞和细胞趋化。通过向趋化膜作为细胞迁移，他们坚持膜，或下降到底层媒体的底面，并随后染色和视觉计算1。在此方法中，细胞暴露在一个陡峭的和短暂的趋化因子梯度，这被认为是2组织中发现的梯度差表示。 另一个检测系统，琼脂糖下的趋化检测，3，4项措施，各地根据琼脂糖凝胶层形成薄薄的水膜，在固体基质细胞运动。梯度在琼脂糖开发的是浅层的，被认为是一个应用程序ropriate代表性的自然发生的梯度。可行驶距离的显微成像评价趋。 Boyden小室法和下琼脂糖检测通常配置为端点检测。 ECIS的/出租车自动化系统相结合的电动细胞基质阻抗遥感（ECIS）的5，6下琼脂糖方法。在这个实验中，目标电极位于8室。一个大型的反电极贯穿每8室（ 图2）。每个室充满琼脂糖和两个小口井是在琼脂糖上削减目标电极的两侧。一个也充满了测试细胞的人口，而其他持有扩散的趋化因子的来源（ 图3）。通过系统的电流可以被用来确定在发生细胞越过目标电极的电阻变化。对靶细胞的焊条é增加阻力系统6。此外，在抵抗快速波动代表在细胞与电极表面相互作用的变化和正在进行的细胞形状变化的指示。 ECIS的/出租车系统可以在多长时间实时测量细胞群体的运动，但也很敏感，足以探测到一个单细胞，在到达目标电极。 被称为粘菌discoidium迁移叶酸梯度7，8 ECIS的/出租车9可以准确测量其趋化反应的存在。白细胞趋化反应到SDF1α趋拮抗剂也被ECIS的10，11 /出租车测量。一个白细胞反应SDF1α的例子如图1所示。

Watch this Scientific Journal Video about Measurement of Cellular Chemotaxis with ECIS/Taxis at JoVE.com

Measurement of Cellular Chemotaxis with ECIS/Taxis

企业家信心指数/出租车系统是一个自动化的，实时的检测，测量细胞趋化。在这个实验中，细胞的琼脂糖层到达目标电极下方移动。细胞运动耐交流电流0开始测量。

ECIS的/出租车细胞趋化测量

Cellular movement in response to external stimuli is fundamental to many cellular processes including wound healing, inflammation and the response to ...

Research

JoVE Journal

Biology

Studies of Bacterial Chemotaxis Using Microfluidics - Interview

使用微流体的细菌的趋化作用的研究 - 访

科研

生物学

A Microfluidic Device with Groove Patterns for Studying Cellular Behavior

微流体装置与Groove的模式，为研究细胞行为

We describe a microfluidic device with microgrooved patterns for studying cellular behavior.  This microfluidic platform consists of a top fluidic channel ...

Making Patch-pipettes and Sharp Electrodes with a Programmable Puller

膜片吸液管和夏普电极与一个可编程的拖轮

Glass microelectrodes (also called pipettes) have been a workhorse of electrophysiology for decades.  Today, such pipettes are made from glass capillaries ...

Identification of protein complexes with quantitative proteomics in S. cerevisiae

鉴定与定量蛋白质组学在酿酒酵母中的蛋白质复合物

Lipids are the building blocks of cellular membranes that function as barriers and in compartmentalization of cellular processes, and recently, as ...

Imaging Exocytosis in Retinal Bipolar Cells with TIRF Microscopy

TIRF显微镜视网膜双极细胞的影像胞外分泌

Total internal reflectance fluorescence (TIRF) microscopy is a technique that allows the study of events happening at the cell membrane, by selective ...

Measurement Of Neuromagnetic Brain Function In Pre-school Children With Custom Sized MEG

在学前儿童Neuromagnetic的脑功能自定义尺寸的MEG测量

Magnetoencephalography is a technique that detects magnetic fields associated with cortical activity [1]. The electrophysiological activity of the brain ...

A Microfluidic Device for Quantifying Bacterial Chemotaxis in Stable Concentration Gradients

一个微流体装置量化细菌趋稳定的浓度梯度

Chemotaxis allows bacteria to approach sources of attractant chemicals or to avoid sources of repellent chemicals. Bacteria constantly monitor the ...

Imaging G-protein Coupled Receptor (GPCR)-mediated Signaling Events that Control Chemotaxis of Dictyostelium Discoideum

Imaging G-protein Coupled Receptor GPCR-mediated Signaling Events that Control Chemotaxis of Dictyostelium Discoideum

成像摹蛋白偶联受体（GPCR）介导的信号，控制趋化粘菌</em

Many eukaryotic cells can detect gradients of chemical signals in their environments and migrate accordingly 1.  This guided cell migration is referred as ...

Measurement of Lifespan in Drosophila melanogaster

Measurement of Lifespan in Drosophila melanogaster

寿命测量<em&gt;果蝇</em

Aging is a phenomenon that results in steady physiological deterioration in nearly all organisms in which it has been examined, leading to reduced ...

C. elegans Chemotaxis Assay

C. elegans Chemotaxis Assay

线虫趋含量的

Many organisms use chemotaxis to seek out food sources, avoid noxious substances, and find mates.  Caenorhabditis elegans has impressive chemotaxis ...