Name: stretching micropatterned cells on a pdms membrane
Uploaded: 2014-01-22T13:45:00
Description: Watch this Scientific Journal Video about Stretching Micropatterned Cells on a PDMS Membrane at JoVE.com

这项工作是由居里研究所，法国巴黎创立。机械担架由达米安屈弗利耶（居里研究所）设计，由GREM制造（机械公司 - grem.com）。 PDMS上的图案是由阿马尔Azioune（波尔多第二大学）开发的。

1。 PDMS的钝化<ol><li>切下一块PDMS大约35毫米×20毫米从预先制成薄片（例如，GelPak，因为在材料的表中列出）。 </li><li>除去塑料的顶部和底部的保护层（如果需要）和使用镊子将PDMS在一个塑料（不是细胞培养物处理的）培养皿。 </li><li>用70％乙醇在30振荡/分洗涤PDMS 5分钟，在旋转器上。 </li><li>通过在其上流动的空气干燥的表面。 </li><li>照亮深紫外光（λ= 180纳米），用于在从约5厘米的紫外线灯?...

<ol>
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虽然这种技术已经被使用无数次，并经过全面的测试，也有可能导致一个失败的实验的几个关键步骤。 关于PDMS： 对于这项工作，GelPak，市售的薄的PDMS片，使用。另外PDMS表可以从PDMS结构直接转换。我们建议使用GelPak，因为它是更具有可重复性，而且不太可能突破相比，定制的PDMS。 关于设备： 在这个协议中使用的拉...

在高等生物组成一个组织中的细胞受到机械张力和拉伸力无论是从外部环境还是从周围细胞1,2到来。细胞必须适应并以维持组织的完整性抵御这些力量。这种力量也是在开发3,4组织形态发生重要。在培养的细胞施加机械力是一种能够模仿一个组织可能会发生什么，但细胞形态和细胞变形5,6的定量和独立控制。为此，一些技术可能被使用。人们可以对细胞按（整个细胞或它的一部分），例如，使用原子力显微镜或衍生物7,8或伸展细胞生长在衬底上。 本文描述的方法演示了如何伸展镀细胞平面基板。在m该技术最初被开发，评估力量的作用发挥itotic哺乳动物细胞9。有丝分裂细胞保持连接到所述衬底通过回缩纤维和拉伸的膜施加的力上的那些纤维，这反过来又引起有丝分裂纺锤体的旋转。结合粘合微图案和伸展的利益，是实现部队和单个细胞的形状的独立控制。它例如可伸展的卵形细胞变成完全各向同性的圆形形状，而单轴拉伸被施加。如果该细胞不制地图上微图案，单轴拉伸导致细胞的伸长，与具有长轴线与拉伸轴一致大多数细胞。它是那么难以分离的长轴对准和施加到所述单元的伸展的效果的效果。 该装置适用于任何活细胞成像，包括长的时间推移的荧光显微镜和药物可在实验过程中被加入。深UV的缩微方法10在Azioune细节被描述等 11上图案化的PDMS被描述在Azioune 等 12本拉伸协议是卡普里等人 13的视频版本

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	<tbody>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">GelPak</td>
			<td style="width:71px;">GelPak</td>
			<td style="width:119px;">PF-60-X4</td>
			<td style="width:167px;">Different thickness/stickiness are available. One alternative could be to cast your PDMS yourself.</td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:216px;">Silicon grease</td>
			<td style="width:71px;">GE Bayer Silicones</td>
			<td style="width:119px;">Baysilone-Paste</td>
			<td>This one is biocompatible</td>
		</tr>
		<tr height="63">
			<td height="63" style="height:63px;width:216px;">Stretching device</td>
			<td style="width:71px;">GREM m&#233;canique</td>
			<td style="width:119px;">Stretcher 2011</td>
			<td></td>
		</tr>
		<tr height="84">
			<td height="84" style="height:84px;width:216px;">EDC (N-(3-Dimethylaminopropyl)-N&#8242;-ethylcarbodiimide hydrochloride)</td>
			<td style="width:71px;">Sigma</td>
			<td align="right" style="width:119px;">3450</td>
			<td>Stable 6 months at -20 &#176;C</td>
		</tr>
		<tr height="63">
			<td height="63" style="height:63px;width:216px;">NHS (N-Hydroxysulfosuccinimide sodium salt)</td>
			<td style="width:71px;">Sigma</td>
			<td align="right" style="width:119px;">56485</td>
			<td>Protect from humidity</td>
		</tr>
		<tr height="63">
			<td height="63" style="height:63px;width:216px;">Pll-g-peg (PLL(20)-g[3.5]-PEG(2) 20 mg)</td>
			<td style="width:71px;">SurfaceSolutions (Zurich)</td>
			<td style="width:119px;"></td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">Synthetic Quartz photomask</td>
			<td style="width:71px;">Toppan</td>
			<td style="width:119px;"></td>
			<td>Take standard binary photomask in Quartz</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">Fibronectin from bovine plasma</td>
			<td style="width:71px;">Sigma</td>
			<td style="width:119px;">F1141</td>
			<td></td>
		</tr>
	</tbody>
</table>

stretching micropatterned cells on a pdms membrane

的机械力作用在细胞和/或组织在许多过程中发挥着重要作用。我们已经开发出一种装置以拉伸细胞接种于聚二甲基硅氧烷（PDMS）膜，具有成像相容。这种技术是可重复的和通用性。将PDMS膜可微图案以限制细胞或组织到一个特定的几何形状。第一步是打印微图案上的PDMS膜用深紫外光的技术。将PDMS膜，然后装上一个机械担架。一室势必与生物相容性脂膜，使拉伸过程中滑翔的顶部。将细胞接种，并使其分散进行数小时的微图案。样品可以被拉伸和未拉伸多次与使用测微螺旋的。这需要不到一分钟的拉伸应用于其完整程度（约30％）。这里提出的技术并不包括机动设备，这是必需的快速和/或计算机控制的拉伸pplying反复伸展周期，但是这是可以实现的。拉伸的细胞或组织，感兴趣的可以用于与细胞的力量，细胞响应机械应力和组织形态发生的问题。此视频演示将展示如何避免可能出现的时候做这一类的看似简单的实验典型问题。

<html>在这个视频协议提出的技术允许在有丝分裂哺乳动物细胞的纤维回缩力的应用。实际上，在细胞分裂期间，在有丝分裂阶段，哺乳动物细胞缩回取的球的形状，并留下由膜包围薄肌动线，它们分别连接到基板上。这些电缆（回缩纤维），是细胞的几何形状在进入分裂的记忆。通过在PDMS薄膜的光掩模制作微图案与深紫外光（ 图1）允许细胞粘附的几何控制。通过拉伸衬底在中期的?...

Watch this Scientific Journal Video about Stretching Micropatterned Cells on a PDMS Membrane at JoVE.com

Stretching Micropatterned Cells on a PDMS Membrane

这个手稿提出了一种技术，使用单向拉伸应用或释放力对粘附细胞或组织中。

在硅橡胶膜拉伸微图案细胞

Mechanical forces exerted on cells and/or tissues  play a major role in numerous processes. We have developed a device to stretch  cells plated on a ...

Research

JoVE Journal

Bioengineering

Micropatterned Surfaces to Study Hyaluronic Acid Interactions with Cancer Cells

Micropatterned表面透明质酸与肿瘤细胞相互作用的研究

Cancer invasion and progression involves a motile cell phenotype, which is under complex regulation by growth factors/cytokines and extracellular matrix ...

科研

生物工程

Improved Visualization and Quantitative Analysis of Drug Effects Using Micropatterned Cells

改进的可视化和定量分析，药物使用Micropatterned细胞的影响

To date, most HCA (High Content Analysis) studies are carried out with adherent cell lines grown on a homogenous substrate in tissue-culture treated ...

Stretching Short Sequences of DNA with Constant Force Axial Optical Tweezers

伸展的DNA短序列恒力轴向光镊

Single-molecule techniques for stretching DNA of contour lengths  less than a kilobase are fraught with experimental difficulties. However, many ...

A Novel Stretching Platform for Applications in Cell and Tissue Mechanobiology

在细胞和组织力学生物学应用一种新型的拉伸平台

Tools that allow the application of mechanical forces to cells and tissues or that can quantify the mechanical properties of biological tissues have ...

Luminescence Resonance Energy Transfer to Study Conformational Changes in Membrane Proteins Expressed in Mammalian Cells

荧光共振能量转移研究构象变化的膜蛋白在哺乳动物细胞中表达

Luminescence Resonance Energy Transfer, or LRET, is a powerful technique used to measure distances between two sites in proteins within the distance range ...

Optimizing Attachment of Human Mesenchymal Stem Cells on Poly(&#949;-caprolactone) Electrospun Yarns

Optimizing Attachment of Human Mesenchymal Stem Cells on PolyÃƒÅ½Ã‚Âµ-caprolactone Electrospun Yarns

优化人骨髓间充质干细胞附着在聚（ε己内酯）静电纺纱线

Research into biomaterials and tissue engineering often includes cell-based in vitro investigations, which require initial knowledge of the starting cell ...

Gene Transfection toward Spheroid Cells on Micropatterned Culture Plates for Genetically-modified Cell Transplantation

基因转染向着文化微模式球体板细胞基因修饰的细胞移植

To improve the therapeutic effectiveness of cell transplantation, a transplantation system of genetically modified, injectable spheroids was developed. ...

Microfluidic Genipin Deposition Technique for Extended Culture of Micropatterned Vascular Muscular Thin Films

微京尼平沉积技术的微模式化血管肌肉薄膜的扩展文化

The chronic nature of vascular disease progression requires the development of experimental techniques that simulate physiologic and pathologic vascular ...

Construction of Modular Hydrogel Sheets for Micropatterned Macro-scaled 3D Cellular Architecture

构建模块化水凝胶片的微模式化宏观缩放3D细胞结构

Hydrogels can be patterned at the micro-scale using microfluidic or micropatterning technologies to provide an in vivo-like three-dimensional (3D) tissue ...

Double Emulsion Generation Using a Polydimethylsiloxane (PDMS) Co-axial Flow Focus Device

Double Emulsion Generation Using a Polydimethylsiloxane PDMS Co-axial Flow Focus Device

复乳代使用聚二甲基硅氧烷（PDMS）同轴流量聚焦设备

Double emulsions are useful in a number of biological and industrial applications in which it is important to have an aqueous carrier fluid. This paper ...