Name: creating two-dimensional patterned substrates for protein and cell confinement
Uploaded: 2011-09-06T13:00:00
Description: Watch this Scientific Journal Video about Creating Two-Dimensional Patterned Substrates for Protein and Cell Confinement at JoVE.com

我们要感谢华盛顿大学的集体知识，这个协议可能在整个毛雷尔组。这项工作的经费是由国家精神卫生研究所（1R01MH085495）。

1。制备的图案硕士（图1） <ol><li>中心硅片上的自旋涂布在两个周期自旋方案的表1中的最初步骤，并用丙酮冲洗晶圆。丙酮蒸发过程中的自旋留下一个清洁，干燥的晶圆计划的第二步。 </li><li>应用约1毫升AZ9245光阻/（直径）晶圆和自旋大衣，使用表1中所述的条件。 </li><li>软烤的光致抗蚀剂涂层的晶圆在110 ° C为2米，用高均匀烤盘。 </li><li> Photopattern基材，使用一个直写光刻系统或光刻系统和适...

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一个用于PDMS的邮票创造主的平版印刷生产中可能出现的一些问题。曝光不足，在若隐若现的模式和曝光过度放大或缺少的功能抵抗涂​​层的晶圆结果抵抗涂层的晶圆结果。在一般情况下，与大特征尺寸（&gt; 10微米）的主人是相对容易的图案和开发，而较小的特点的主人可以要求广泛photopatterning和发展参数优化（超越光阻制造商推荐的参数）。最困难的主人，生产大型和小型的功能结合起来。 <...

<table border="1"><tbody><tr><th>试剂名称</th><th>公司</th><th>目录编号</th><th>评论（可选） </th></tr><tr><td>硅片</td><td>晶圆回收服务</td><td></td><td> 2英寸</td></tr><tr><td>旋转涂布机/热板</td><td>布鲁尔科技</td><td> CEE 200CB旋转烘烤系统</td><td></td></tr><tr><td> AZ9245光刻胶</td><td>梅斯化工公司</td><td> 105880034-1160 </td><td></td></tr><tr><td>直写光刻系统</td><td>迈特SRL </td><td> LW325 LaserWriter系统</td><td></td></tr><tr><td>掩模对准器</td><td> HTG </td><td> 3小时</td><td></td></tr><tr><td> AZ 400K开发</td><td>梅斯化工公司</td><td> 105880018-1160 </td><td></td></tr><tr><td> Sylgard 182有机硅弹性体套件</td><td>道康宁</td><td></td><td></td></tr><tr><td>没有25毫米。 1圆形盖玻片</td><td> VWR </td><td> 16004-310 </td><td></td></tr><tr><td>等离子体氧化</td><td>迪纳</td><td>飞秒</td><td></td></tr><tr><td>钛片Kamis </td><td>注册成立</td><td></td><td> 99.95％的纯</td></tr><tr><td>黄金颗粒</td><td> Kamis团</td><td></td><td> 99.999％的纯</td></tr><tr><td>电子束蒸发</td><td>库尔特J. Lesker </td><td> 75物理气相沉积薄膜沉积系统</td><td>电子束配件</td></tr><tr><td> Hexadecanethiol </td><td>阿法埃莎</td><td> A11362 </td><td></td></tr><tr><td> 1，mercaptoundec - 11 - YL）TETRA（乙二醇） </td><td>西格玛爱秩序</td><td> 674508 </td><td></td></tr><tr><td>乙醇</td><td> Pharmco - aaper </td><td> 111000200 </td><td> 200证明，绝对</td></tr><tr><td>封口膜</td><td> VWR </td><td> 52858-000 </td><td></td></tr><tr><td> DPBS </td><td> VWR </td><td> 4500-434 </td><td>如果没有钙和镁</td></tr><tr><td>鼠标层粘连我</td><td> VWR </td><td> 95036-762 </td><td></td></tr><tr><td>人类血浆纤维连接蛋白</td><td> Invitrogen公司</td><td> 33016-015 </td><td></td></tr><tr><td> AlexaFluor ® 647羧酸琥珀酰亚胺酯</td><td> Invitrogen公司</td><td> A - 20006 </td><td></td></tr><tr><td> MitoTracker红580 </td><td> Invitrogen公司</td><td> M22425 </td><td></td></tr><tr><td> AlexaFluor ® 350羧酸琥珀酰亚胺酯</td><td> Invitrogen公司</td><td> A - 10168 </td><td></td></tr><tr><td>抗层粘连蛋白抗体</td><td> Fisher Scientific则</td><td> AB2034MI </td><td></td></tr></tbody></table>

creating two-dimensional patterned substrates for protein and cell confinement

微接触印刷提供了一个快速，为创建良好定义的图案衬底的高度重复性的方法1虽然微接触印刷，可直接打印大量的分子，包括蛋白质，DNA的 2，3和硅烷，形成自我金长链烷烃硫醇组装单层膜（SAMS）提供了一个简单的方法来限制​​蛋白质和细胞含有粘合剂和耐地区的具体模式。这禁闭可用于控制细胞的形态，是用于研究的蛋白质和细胞生物学的各种问题。在这里，我们描述了细胞研究创造良好定义的蛋白质模式的一般方法，5这个过程包括三个步骤：一个图案主生产采用光刻，创造一个PDMS邮票，和微接触印刷金涂基材。图案后，这些细胞培养基质围蛋白质和/或细胞（原代细胞或细胞株）的格局。 对图案的蛋白质/细胞粘附地区和非粘性地区的精确控制允许使用的自组装单层化学，不能使用直接的蛋白质冲压实现。 Hexadecanethiol，微接触印刷步骤所用的长链烷烃硫醇，产生一个疏水表面容易吸附溶液中的蛋白质。乙二醇终止的巯基，用于回填基板的非打印区域，创建一个单层抗蛋白质吸附，因此，细胞的生长。6这些巯基的单体生产的高度结构化膜，精确地界定地区的基板，可以支持蛋白质吸附和细胞的生长。因此，这些承印物是有用的各种广泛的应用，从7间行为研究创造微电子8。 虽然已用于细胞培养的研究，包括从本集团的工作，创建模式，直接在玻璃基板上使用trichlorosilanes的其他类型的单层化学，9图案形成单分子膜对黄金的烷烃硫醇直截了当的准备。此外，单层准备使用的单体商业，​​稳定，不需要储存或处理惰性气氛下。烷烃硫醇准备的图案基板也可以被回收和重复使用多次，维持细胞禁闭10。

Watch this Scientific Journal Video about Creating Two-Dimensional Patterned Substrates for Protein and Cell Confinement at JoVE.com

Creating Two-Dimensional Patterned Substrates for Protein and Cell Confinement

金长链烷烃硫醇形成的自组装单层膜（SAMS）提供了定义良好的蛋白质模式的形成和细胞禁闭基板。使用一个邮票回填与聚二甲基硅氧烷（PDMS）微接触印刷hexadecanethiol乙二醇终止烷烃硫醇单体生产蛋白质和细胞吸附加盖hexadecanethiol地区唯一的模式。

蛋白质和细胞坐月子创建二维图案衬底

Microcontact printing provides a rapid, highly reproducible method for the creation of well-defined patterned substrates.1  While microcontact printing ...

Research

JoVE Journal

Bioengineering

Creating Transient Cell Membrane Pores Using a Standard Inkjet Printer

创建瞬态细胞膜毛孔，使用标准的喷墨打印机

Bioprinting has a wide range of applications and significance, including tissue engineering, direct cell application therapies, and biosensor ...

科研

生物工程

Creating Adhesive and Soluble Gradients for Imaging Cell Migration with Fluorescence Microscopy

创建的胶粘剂和可溶性梯度荧光显微镜成像细胞迁移

Cells can sense and migrate towards higher concentrations of adhesive cues such as the glycoproteins of the extracellular matrix and soluble cues such as ...

Cell Patterning on Photolithographically Defined 
Parylene-C: SiO2 Substrates

Cell Patterning on Photolithographically Defined 
Parylene-C: SiO2 Substrates

在光刻技术定义的聚对二甲苯-C细胞图案：二氧化硅<sub&gt; 2</sub&gt;基板

Cell patterning platforms support broad research goals, such as construction of predefined in vitro neuronal networks and the exploration of certain ...

Ordering Single Cells and Single Embryos in 3D Confinement: A New Device for High Content Screening

订货单细胞和单胚胎的三维约束：高内涵筛选新设备

Biological cells are usually observed on flat (2D) surfaces. This condition is not physiological, and phenotypes and shapes are highly variable. Screening ...

Fluorescence Recovery after Merging a Droplet to Measure the Two-dimensional Diffusion of a Phospholipid Monolayer

合并一个液滴后荧光恢复来衡量磷脂单层的二维扩散

We introduce a new method to measure the lateral diffusivity of a surfactant monolayer at the fluid-fluid interface, called fluorescence recovery after ...

Expanding Nanopatterned Substrates Using Stitch Technique for Nanotopographical Modulation of Cell Behavior

扩大奈米基材使用缝合技术的细胞行为Nanotopographical调制

Substrate nanotopography has been shown to be a potent modulator of cell phenotype and function. To dissect nanotopography modulation of cell behavior, a ...

High-resolution Patterned Biofilm Deposition Using pDawn-Ag43

使用 pDawn-Ag43 的高分辨率图案生物膜沉积

Spatial structure and patterning play an important role in bacterial biofilms. Here we demonstrate an accessible method for culturing E. coli biofilms ...

Evaluation of Keratinocyte Proliferation on Two- and Three-dimensional Type I Collagen Substrates

二维和三维 i 型胶原蛋白基质角质形成细胞增殖的评价

Type I collagen, useful as a substrate for cell culture, exists in two forms: the two-dimensional, non-fibrous form and three-dimensional, fibril form. ...

Patterning the Geometry of Human Embryonic Stem Cell Colonies on Compliant Substrates to Control Tissue-Level Mechanics

在符合标准的基质上对人类胚胎干细胞菌落的几何结构进行模式化，以控制组织级力学

Human embryonic stem cells demonstrate a unique ability to respond to morphogens in vitro by self-organizing patterns of cell fate specification that ...

Preparing Protein Producing Synthetic Cells using Cell Free Bacterial Extracts, Liposomes and Emulsion Transfer

使用无细胞细菌提取物、脂质体和乳液转移制备蛋白质生成合成细胞

The bottom-up assembly approach for construction of synthetic cells is an effective tool for isolating and investigating cellular processes in a cell ...