Name: multiscale structures aggregated by imprinted nanofibers for functional surfaces
Uploaded: 2018-09-11T14:45:00.000+00:00
Duration: 6 min 14 s
Description: Watch this Scientific Journal Video about Multiscale Structures Aggregated by Imprinted Nanofibers for Functional Surfaces at JoVE.com

这一材料的基础是通过由科学、信息和通信技术和未来规划部 (NRF-2017R1A2B4008053) 和贸易、工业和能源部 (NRF) 资助的韩国国家研究基金会为基础科学研究项目提供支助的工作 (MOTIE, 韩国) 根据工业技术创新计划10052802号和韩国技术进步研究所 (永吉), 通过鼓励方案为工业的经济合作区 (N0002310)。

1. 利用氧化铝过滤器制备纳米微尺度结构表面 (图 1)
<ol><li>购买一个氧化铝过滤器的孔径大小, 高度, 直径为200毫微米, 60 µm, 和25毫米, 分别。</li>
	<li>1.2. 清洗具有100微米厚度的聚对苯二甲酸乙二醇酯 (PET) 膜的表面, 使用丙酮与99.8% 和异丙醇 (IPA) 99.9% 为5分钟, 并使用气枪完全干燥3分钟。</li>
	<li>将 PET 薄膜放置在无污染物的平坦表面上, 并添加0.1 毫升的 UV 固化聚氨酯丙烯酸酯 (PUA) 型树脂, 其粘度为 257.4 cPs 到表面。</li>
	<li>将阳极氧化铝过滤器放在树脂上, 均匀压匀, 使用直径为32毫米的橡皮滚筒。树脂的传播在视觉上被证实, 因此必须反复地和仔细地推挤在压下的滚筒。 注意: 阳极氧化铝过滤器是脆的, 如果施加过多的作用力, 可能会断裂。</li>
	<li>轧制后, 将与 PET 和氧化铝过滤器 (附使用树脂) 制成的试样暴露于三十年代的波长为 365 nm 的紫外光以固化树脂。</li>
	<li>将固化试样浸泡在100毫升2米氢氧化钠溶液中, 10 分钟即可溶解过滤器。 注: SEM 图像显示结构的横断面和表面 (图 2)。</li>
	<li>用空气枪清洗试样, 然后用气枪完全干燥3分钟。 注: EDX 分析证实 Na 和 Al 未被检测到并被完全蚀刻 (图 3)。</li>
</ol>2. 表面处理
<ol><li>紫外线臭氧处理</li>
	<li>用纳米微尺度结构清洁试样, 使用 IPA 和 DI 水5分钟, 然后用气枪干3分钟。</li>
	<li>使用 UV 射线 (波长为 185-254 nm) 照射到试样多尺度结构的一侧 (侧面有多尺度结构), 60 分钟。 注: 紫外线臭氧设备的强度为25兆瓦/厘米2。</li>
	<li>Octadecyltrichlorosilane 自组装</li>
	<li>在手套箱内放置一个热板, 并保持一个2的环境, 用于蒸气沉积过程。</li>
	<li>用胶带将试样的边缘固定在玻璃或平板上。确保玻璃或板材的尺寸足够大, 足以覆盖烧杯的顶部 (直径为8毫米, 高度为13毫米)。</li>
	<li>将烧杯放在 5 "x7" 的热板上, 用吸管在烧杯中加入2毫升的溶液。</li>
	<li>用玻璃或板面覆盖烧杯, 试样朝下进入烧杯。</li>
	<li>处理60分钟在100摄氏度, 然后从手套盒中取出标本。 注: 在涂布工艺后, 必须清洗烧杯和手套盒。</li>
</ol>3. 注射润滑剂制备功能表面
<ol><li>将大约0.2 毫升 的氟碳 (PFC) 液体存入涂覆的自聚集碳纤维组件上。</li>
	<li>用物镜的光学显微镜观察 PFC 的润湿过程, 5 x-20。</li>
	<li>将样品放置在垂直位置几个小时, 卸下多余的 PFC 液体。</li>
</ol>

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作者没有竞争的金融利益来披露。

制备自聚集碳纤维总成的关键步骤是确保在胶辊上应用树脂时, 脆性氧化铝过滤器不会断裂。事实上, 应该确保氧化铝过滤器在蚀刻步骤之前的任何一点都不会断裂。由于氧化铝过滤器直径为25毫米, 基体的尺寸约为 30 x 30 毫米。
自聚集碳纤维组件允许我们通过适当的表面处理提供各种功能表面。在印迹后, 原表面是亲水性的, 但经涂膜后, 受紫外线臭氧处理和表面能量的变化, 可对其进行改性和疏水性。此外, 所提出的多尺度孔隙结构可以通过液体润滑剂注入过程转化为光滑的表面。
纳米微尺度结构的表面是不透明的, 可能是由于聚合纤维的不规则性, 这种特性可以在光学应用中使用。因此, 在随后的研究中, 我们将利用紫外-可见光谱仪研究基体的光学特性。我们期望这种表面的光学特性可以应用到需要光反射的工业中。

纳米颗粒、纳米管和纳米纤维等纳米结构已引起科学界的关注, 因为它们在各种应用中表现出独特的特性, 包括电子、生物医学、光学和表面工程1,2,3,4,5,6,7,8。特别是, 纳米纤维广泛应用于可伸缩和透明电极9, 耐磨传感器10,11, 互连12,13, 和纳米光学应用14. 在制造纳米结构的各种方法中, 如溶胶-凝胶方法、自组装、光刻和复制15、16、17、18, 19、20, 使用模板进行直接复制目前被认为是一种有希望的方法, 因为它简单、经济高效, 适用于各种可固化材料21、22,23,24,25,26。
由于其多尺度结构具有大量的纳米孔隙和微尺度高度, 因此, 氧化铝被广泛用作制备纳米纤维和纳米管的模板, 具有高长宽比27、28、29,30. 然而, 由于表面张力在如此高的纵横比, 纤维容易地聚集31,32,33。现有的研究已经证明, 长宽比大于15:1 的纳米纤维不直立, 而是聚合, 而比不到5:1 的人则单独隔离, 没有聚集33,34。毛细管力和表面张力对氧化铝的去除起着重要的作用, 蚀刻是碳纤维制造过程中的工艺之一。当长宽比增加时, 纤维间的表面张力会使它们相互靠近, 导致聚集。一些研究集中在防止这种聚合的方法35, 这在聚合物和金属纳米纤维中特别被观察到。其中, 碳纤维表面的水化可以减少团聚, 因为当液体占据纳米纤维之间的空隙时, 表面张力就会减小。此外, 冻干法还可以降低纤维间的表面张力, 从而减少聚集。然而, 尽管作出了各种努力, 高纵横比的纳米纤维的矫直仍然是一个挑战。
为此, 我们报告了一种独特的方法来制造复杂的碳纤维的多尺度结构, 利用聚集现象的积极方式。在这里, 碳纤维结构是用氧化铝过滤器和聚氨酯丙烯酸酯 (PUA) 型树脂, 粘度为 257.4 cP。在 uv 纳米压印 (uv-零) 进行后, 模具用氢氧化钠溶液蚀刻。为了表征所提出的多尺度结构, 我们研究了聚合纳米纤维样品的模式行为和表面润湿性, 如自组装单层涂层和紫外线臭氧处理等表面处理。.此外, 我们建议, 多孔表面可以简单地转化为一个光滑的表面使用润滑剂注入过程。

<table><tbody><tr><td>MINS 511RM</td><td>Minuta Tech</td><td></td><td>UV curable resin</td></tr><tr><td>Octadecyltrichlorosilane (OTS)</td><td>Aldrich</td><td></td><td>Surface treatment</td></tr><tr><td>Sodium oxidanide</td><td>SAMCHUN</td><td></td><td>Etching solution</td></tr><tr><td>Anopore Inoganic Membranes</td><td>Whatman</td><td></td><td>25mm/0.2&amp;micro;m</td></tr><tr><td>MT-UV-A 47</td><td>Meiji Techno</td><td></td><td>UV curing equipment</td></tr><tr><td>UVC-30</td><td>Jaesung Engineering</td><td></td><td>UVO treatment equipment</td></tr><tr><td>Smart Drop Plus</td><td>FEMTOFAB</td><td></td><td>Contact angle measurement</td></tr><tr><td>Fluorinert FC-70</td><td>3M</td><td></td><td>liquid mixture of completely fluorinated aliphatic compounds</td></tr><tr><td>Polyethylene terephthalate film</td><td>Sunchem</td><td></td><td>Substrate</td></tr><tr><td>Acetone (99.8%)</td><td>Daejung</td><td></td><td>Cleaning solution</td></tr><tr><td>Isopropyl alcohol (99.9%)</td><td>Daejung</td><td></td><td>Cleaning solution</td></tr><tr><td>Rubber roller</td><td>Hwahong</td><td></td><td>For application of resin</td></tr><tr><td>Corning Stirring Hot Plates</td><td>Corning</td><td></td><td>Hot plate equipment (5&quot; x 7&quot;)</td></tr></tbody></table>

multiscale structures aggregated by imprinted nanofibers for functional surfaces

多尺度表面结构引起了越来越多的兴趣, 因为一些潜在的应用在表面设备。然而, 目前在这一领域存在的一个挑战是采用轻便、经济高效和高通量的方法制造混合微纳米结构。为了克服这些挑战, 本文提出了一种用阳极氧化铝 (氧化铝) 过滤器和纳米纤维的蒸发自聚过程来制造多尺度结构的协议。不像以前试图理顺纳米纤维的尝试, 我们展示了一种独特的制造方法, 多尺度聚合纳米纤维的高纵横比。此外, 研究了这些结构在各种液体中的表面形貌和润湿性, 以利于它们在多功能表面的应用。

用氧化铝过滤器作为印迹模, 演示了一种快速、简便的制备多尺度纳米微混合结构的方法。整个过程花费了30分钟 (图 4)。据指出, 在使用氢氧化钠进行蚀刻过程后, 由于表面张力引起的聚集碳纤维装配, 结果表面呈现出与原氧化铝过滤器相似的不透明颜色。进一步, EDX 分析的结果证实了氧化铝过滤器被湿化学蚀刻完全去除 (图 3)。
通过将接触角滴入水滴 5 ul 的试样表面来测定其表面特征。由于在氧化铝过滤介导的压印过程中所使用的材料呈现 superhydrophilicity, 而由于自聚合的纳米纤维, 制备的多尺度结构具有高孔隙网络, 水滴往往是瞬间吸收到基体。然而, 亲水性可以改成疏水性使用适当的表面处理。如图 5所示, 我们展示了印迹多尺度结构的表面被改成一个疏水性表面, 在涂层后, 接触角近似117°。此外, 紫外线臭氧处理可以进一步增加表面的接触角约 10° (图 6)。在对印迹表面进行了连续涂膜和紫外线臭氧处理后, 证实了所产生的接触角增加到 134° (图 7)。
涂层试样的表面和截面显示了纳米纤维的聚合 (图 5), 从而产生了一个酒窝结构。这种凹窝结构的大小和方向不规则;然而, 这种现象发生在整个标本表面。试样表面在受到紫外线臭氧处理过程36后变得平滑 (图 6和 图 7)。这也是为什么在紫外线臭氧处理过程后, 表面接触角增加的原因。该现象在试样表面均匀发生, 接触角误差小于3°。
<img alt="Figure 1" class="xfigimg" src="/files/ftp_upload/58356/58356fig1.jpg"> 图 1: 用可溶性氧化铝制造结构的程序.<a href="https://www.jove.com/files/ftp_upload/58356/58356fig1large.jpg" target="_blank">请单击此处查看此图的较大版本.</a>
<img alt="Figure 2" class="xfigimg" src="/files/ftp_upload/58356/58356fig2.jpg"> 图 2: SEM 图像的纳米微尺度结构在蚀刻后的过程中, 显示了表面和剖面.<a href="https://www.jove.com/files/ftp_upload/58356/58356fig2large.jpg" target="_blank">请单击此处查看此图的较大版本.</a>
<img alt="Figure 3" class="xfigimg" src="/files/ftp_upload/58356/58356fig3.jpg"> 图 3: 用氧化铝过滤器制造的纳米微尺度结构蚀刻后的 EDX 分析结果.<a href="https://www.jove.com/files/ftp_upload/58356/58356fig3large.jpg" target="_blank">请单击此处查看此图的较大版本.</a>
<img alt="Figure 4" class="xfigimg" src="/files/ftp_upload/58356/58356fig4.jpg"> 图 4: 完全蚀刻后的纳米纤维的聚合制备奈米微尺度结构的示意图.<a href="https://www.jove.com/files/ftp_upload/58356/58356fig4large.jpg" target="_blank">请单击此处查看此图的较大版本.</a>
<img alt="Figure 5" class="xfigimg" src="/files/ftp_upload/58356/58356fig5.jpg"> 图 5: 涂层表面和纳米微结构后的接触角.<a href="https://www.jove.com/files/ftp_upload/58356/58356fig5large.jpg" target="_blank">请单击此处查看此图的较大版本.</a>
<img alt="Figure 6" class="xfigimg" src="/files/ftp_upload/58356/58356fig6.jpg"> 图 6: UV 臭氧处理后的接触角表面和纳米微结构.<a href="https://www.jove.com/files/ftp_upload/58356/58356fig6large.jpg" target="_blank">请单击此处查看此图的较大版本.</a>
<img alt="Figure 7" class="xfigimg" src="/files/ftp_upload/58356/58356fig7.jpg"> 图 7: 在表面和纳米微结构上连续执行涂层和紫外线臭氧处理后的接触角.<a href="https://www.jove.com/files/ftp_upload/58356/58356fig7large.jpg" target="_blank">请单击此处查看此图的较大版本.</a>

Watch this Scientific Journal Video about Multiscale Structures Aggregated by Imprinted Nanofibers for Functional Surfaces at JoVE.com

Multiscale Structures Aggregated by Imprinted Nanofibers for Functional Surfaces

功能表面用印迹纳米纤维聚合的多尺度结构

本文介绍了一种利用阳极氧化铝过滤器制备纳米微尺度结构的简易方法--功能表面。

Multiscale surface structures have attracted increasing interest owing to several potential applications in surface devices. However, an existing ...

multiscale-structures-aggregated-imprinted-nanofibers-for-functional

Nanyang Technological University

Research

JoVE Journal

Engineering

6.5K 次观看.  Changwon National University. 本文介绍了一种利用阳极氧化铝过滤器制备纳米微尺度结构的简易方法--功能表面。

视频: Multiscale Structures Aggregated by Imprinted Nanofibers for Functional Surfaces

Nanomoulding of Functional Materials, a Versatile Complementary Pattern Replication Method to Nanoimprinting

We describe a nanomoulding technique which allows low-cost nanoscale patterning of functional materials, materials stacks and full devices. Nanomoulding combined with layer transfer enables the replication of arbitrary surface patterns from a master structure onto the functional material. Nanomoulding can be performed on any nanoimprinting setup and can be applied to a wide range of materials and deposition processes. In particular we demonstrate the fabrication of patterned transparent zinc oxide electrodes for light trapping applications in solar cells.

We describe a nanomoulding technique which allows low-cost nanoscale patterning of functional materials, materials stacks and full devices. Nanomoulding can be performed on any nanoimprinting setup and can be applied to a wide range of materials and deposition processes.

Nanomoulding功能材料，灵活的互补模式的复制方法，以奈米

We describe a nanomoulding technique which  allows low-cost nanoscale patterning of functional materials, materials stacks  and full devices. Nanomoulding ...

科研

工程学

Manufacturing of Three-dimensionally Microstructured Nanocomposites through Microfluidic Infiltration

Microstructured composite beams reinforced with complex three-dimensionally (3D) patterned nanocomposite microfilaments are fabricated via nanocomposite in&#64257;ltration of 3D interconnected microfluidic networks. The manufacturing of the reinforced beams begins with the fabrication of microfluidic networks, which involves layer-by-layer deposition of fugitive ink filaments using a dispensing robot, filling the empty space between filaments using a low viscosity resin, curing the resin and finally removing the ink.&#160;Self-supported 3D structures with other geometries and many layers (e.g.&#160;a few hundreds layers) could be built using this method. The resulting tubular micro&#64258;uidic networks are then infiltrated with thermosetting nanocomposite suspensions containing nanofillers (e.g.&#160;single-walled carbon nanotubes), and subsequently cured. The infiltration is done by applying a pressure gradient between two ends of the empty network (either by applying a vacuum or vacuum-assisted microinjection). Prior to the infiltration, the nanocomposite suspensions are prepared by dispersing nanofillers into polymer matrices using ultrasonication and three-roll mixing methods. The nanocomposites (i.e.&#160;materials infiltrated) are then solidified under UV exposure/heat cure, resulting in a 3D-reinforced composite structure. The technique presented here enables the design of functional nanocomposite macroscopic products for microengineering applications such as actuators and sensors.

Three-dimensional (3D) microstructured composite beams are fabricated through the directed and localized infiltration of nanocomposites into 3D porous microfluidic networks. The flexibility of this manufacturing method enables the utilization of different thermosetting materials and nanofillers in order to achieve a variety of functional 3D reinforced nanocomposite macroscopic products.

通过微流体渗透制备三维微结构纳米复合材料的制造

Microstructured composite beams reinforced with complex three-dimensionally (3D) patterned nanocomposite microfilaments are fabricated via nanocomposite ...

化学

ECM Protein Nanofibers and Nanostructures Engineered Using Surface-initiated Assembly

The extracellular matrix (ECM) in tissues is synthesized and assembled by cells to form a 3D fibrillar, protein network with tightly regulated fiber diameter, composition and organization. In addition to providing structural support, the physical and chemical properties of the ECM play an important role in multiple cellular processes including adhesion, differentiation, and apoptosis. In vivo, the ECM is assembled by exposing cryptic self-assembly (fibrillogenesis) sites within proteins. This process varies for different proteins, but fibronectin (FN) fibrillogenesis is well-characterized and serves as a model system for cell-mediated ECM assembly. Specifically, cells use integrin receptors on the cell membrane to bind FN dimers and actomyosin-generated contractile forces to unfold and expose binding sites for assembly into insoluble fibers. This receptor-mediated process enables cells to assemble and organize the ECM from the cellular to tissue scales. Here, we present a method termed surface-initiated assembly (SIA), which recapitulates cell-mediated matrix assembly using protein-surface interactions to unfold ECM proteins and assemble them into insoluble fibers. First, ECM proteins are adsorbed onto a hydrophobic polydimethylsiloxane (PDMS) surface where they partially denature (unfold) and expose cryptic binding domains. The unfolded proteins are then transferred in well-defined micro- and nanopatterns through microcontact printing onto a thermally responsive poly(N-isopropylacrylamide) (PIPAAm) surface. Thermally-triggered dissolution of the PIPAAm leads to final assembly and release of insoluble ECM protein nanofibers and nanostructures with well-defined geometries. Complex architectures are possible by engineering defined patterns on the PDMS stamps used for microcontact printing. In addition to FN, the SIA process can be used with laminin, fibrinogen and collagens type I and IV to create multi-component ECM nanostructures. Thus, SIA can be used to engineer ECM protein-based materials with precise control over the protein composition, fiber geometry and scaffold architecture in order to recapitulate the structure and composition of the ECM in vivo.

A method to obtain nanofibers and complex nanostructures from single or multiple extracellular matrix proteins is described. This method uses protein-surface interactions to create free-standing protein-based materials with tunable composition and architecture for use in a variety of tissue engineering and biotechnology applications.

细胞外基质蛋白纳米纤维和纳米结构工程采用表面引发大会

The extracellular matrix (ECM) in tissues is synthesized and assembled by cells to form a 3D fibrillar, protein network with tightly regulated fiber ...

生物工程

Fabricating Reactive Surfaces with Brush-like and Crosslinked Films of Azlactone-Functionalized Block Co-Polymers

In this paper, fabrication methods that generate novel surfaces using the azlactone-based block co-polymer, poly (glycidyl methacrylate)-block-poly (vinyl dimethyl azlactone) (PGMA-b-PVDMA), are presented. Due to the high reactivity of azlactone groups towards amine, thiol, and hydroxyl groups, PGMA-b-PVDMA surfaces can be modified with secondary molecules to create chemically or biologically functionalized interfaces for a variety of applications. Previous reports of patterned PGMA-b-PVDMA interfaces have used traditional top-down patterning techniques that generate non-uniform films and poorly controlled background chemistries. Here, we describe customized patterning techniques that enable precise deposition of highly uniform PGMA-b-PVDMA films in backgrounds that are chemically inert or that have biomolecule-repellent properties. Importantly, these methods are designed to deposit PGMA-b-PVDMA films in a manner that completely preserves azlactone functionality through each processing step. Patterned films show well-controlled thicknesses that correspond to polymer brushes (~90 nm) or to highly crosslinked structures (~1-10 &#956;m). Brush patterns are generated using either the parylene lift-off or interface directed assembly methods described and are useful for precise modulation of overall chemical surface reactivity by adjusting either the PGMA-b-PVDMA pattern density or the length of the VDMA block. In contrast, the thick, crosslinked PGMA-b-PVDMA patterns are obtained using a customized micro-contact printing technique and offer the benefit of higher loading or capture of secondary material due to higher surface area to volume ratios. Detailed experimental steps, critical film characterizations, and trouble-shooting guides for each fabrication method are discussed.

Surface fabrication methods for patterned deposition of nanometer thick brushes or micron thick, crosslinked films of an azlactone block co-polymer are reported. Critical experimental steps, representative results, and limitations of each method are discussed. These methods are useful for creating functional interfaces with tailored physical features and tunable surface reactivity.

Azlactone 功能块共聚物的刷状和交联膜制备反应表面

In this paper, fabrication methods that generate novel surfaces using the azlactone-based block co-polymer, poly (glycidyl methacrylate)-block-poly (vinyl ...

Electrospun Nanofiber Scaffolds with Gradations in Fiber Organization

The goal of this protocol is to report a simple method for generating nanofiber scaffolds with gradations in fiber organization and test their possible applications in controlling cell morphology/orientation. Nanofiber organization is controlled with a new fabrication apparatus that enables the gradual decrease of fiber organization in a scaffold. Changing the alignment of fibers is achieved through decreasing deposition time of random electrospun fibers on a uniaxially aligned fiber mat. By covering the collector with a moving barrier/mask, along the same axis as fiber deposition, the organizational structure is easily controlled. For tissue engineering purposes, adipose-derived stem cells can be seeded to these scaffolds. Stem cells undergo morphological changes as a result of their position on the varied organizational structure, and can potentially differentiate into different cell types depending on their locations. Additionally, the graded organization of fibers enhances the biomimicry of nanofiber scaffolds so they more closely resemble the natural orientations of collagen nanofibers at tendon-to-bone insertion site compared to traditional scaffolds. Through nanoencapsulation, the gradated fibers also afford the possibility to construct chemical gradients in fiber scaffolds, and thereby further strengthen their potential applications in fast screening of cell-materials interaction and interfacial tissue regeneration. This technique enables the production of continuous gradient scaffolds, but it also can potentially produce fibers in discrete steps by controlling the movement of the moving barrier/mask in a discrete fashion.

Here, we present a protocol to fabricate electrospun nanofiber scaffolds with gradated organization of fibers and explore their applications in regulating cell morphology/orientation. Gradients with regard to physical and chemical properties of the nanofiber scaffolds offer a wide variety of applications in the biomedical field.

静电纳米纤维支架与渐变的纤维组织

The goal of this protocol is to report a simple method for generating nanofiber scaffolds with gradations in fiber organization and test their possible ...

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

Substrate nanotopography has been shown to be a potent modulator of cell phenotype and function. To dissect nanotopography modulation of cell behavior, a large area of nanopatterned substrate is desirable so that enough cells can be cultured on the nanotopography for subsequent biochemical and molecular biology analyses. However, current nanofabrication techniques have limitations to generate highly defined nanopatterns over a large area. Herein, we present a method to expand nanopatterned substrates from a small, highly defined nanopattern to a large area using stitch technique. The method combines multiple techniques, involving soft lithography to replicate poly(dimethylsiloxane) (PDMS) molds from a well-defined mold, stitch technique to assemble multiple PDMS molds to a single large mold, and nanoimprinting to generate a master mold on polystyrene (PS) substrates. With the PS master mold, we produce PDMS working substrates and demonstrate nanotopographical modulation of cell spreading. This method provides a simple, affordable yet versatile avenue to generate well-defined nanopatterns over large areas, and is potentially extended to create micro-/nanoscale devices with hybrid components.

A protocol for producing a large area of nanopatterned substrate from small nanopatterned molds for study of nanotopographical modulation of cell behavior is presented.

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 ...

Fabrication of a Multiplexed Artificial Cellular MicroEnvironment Array

Cellular microenvironments consist of a variety of cues, such as growth factors, extracellular matrices, and intercellular interactions. These cues are well orchestrated and are crucial in regulating cell functions in a living system. Although a number of researchers have attempted to investigate the correlation between environmental factors and desired cellular functions, much remains unknown. This is largely due to the lack of a proper methodology to mimic such environmental cues in vitro, and simultaneously test different environmental cues on cells. Here, we report an integrated platform of microfluidic channels and a nanofiber array, followed by high-content single-cell analysis, to examine stem cell phenotypes altered by distinct environmental factors. To demonstrate the application of this platform, this study focuses on the phenotypes of self-renewing human pluripotent stem cells (hPSCs). Here, we present the preparation procedures for a nanofiber array and the microfluidic structure in the fabrication of a Multiplexed Artificial Cellular MicroEnvironment (MACME) array. Moreover, overall steps of the single-cell profiling, cell staining with multiple fluorescent markers, multiple fluorescence imaging, and statistical analyses, are described.

This article describes the detailed methodology to prepare a Multiplexed Artificial Cellular MicroEnvironment (MACME) array for high-throughput manipulation of physical and chemical cues mimicking in vivo cellular microenvironments and to identify the optimal cellular environment for human pluripotent stem cells (hPSCs) with single-cell profiling.

多路人工细胞微环境阵列的制备

Cellular microenvironments consist of a variety of cues, such as growth factors, extracellular matrices, and intercellular interactions. These cues are ...

Fabrication of Gradient Nanopattern by Thermal Nanoimprinting Technique and Screening of the Response of Human Endothelial Colony-forming Cells

Nanotopography can be found in various extracellular matrices (ECMs) around the body and is known to have important regulatory actions upon cellular reactions. However, it is difficult to determine the relation between the size of a nanostructure and the responses of cells owing to the lack of proper screening tools. Here, we show the development of reproducible and cost-effective gradient nanopattern plates for the manipulation of cellular responses. Using anodic aluminum oxide (AAO) as a master mold, gradient nanopattern plates with nanopillars of increasing diameter ranges [120-200&#8239;nm (GP 120/200), 200-280&#8239;nm (GP 200/280), and 280-360&#8239;nm (GP 280/360)] were fabricated by a thermal imprinting technique. These gradient nanopattern plates were designed to mimic the various sizes of nanotopography in the ECM and were used to screen the responses of human endothelial colony-forming cells (hECFCs). In this protocol, we describe the step-by-step procedure of fabricating gradient nanopattern plates for cell engineering, techniques of cultivating hECFCs from human peripheral blood, and culturing hECFCs on nanopattern plates.

Here, we present a protocol for the fabrication of gradient nanopattern plates via thermal nanoimprinting and the method of screening responses of human endothelial progenitor cells to the nanostructures. By using the described technology, it is possible to produce a scaffold that can manipulate cell behavior by physical stimuli.

热 Nanoimprinting 技术制备梯度 Nanopattern 及其对人血管内皮细胞菌落形成的响应筛选

Nanotopography can be found in various extracellular matrices (ECMs) around the body and is known to have important regulatory actions upon cellular ...

Procedure for Fabricating Biofunctional Nanofibers

Electrospinning is an effective processing method for preparing nanofibers decorated with functional groups. Nanofibers decorated with functional groups may be utilized to study material-biomarker interactions i.e. act as biosensors with potential as single molecule detectors. We have developed an effective approach for preparing functional polymers where the functionality has the capacity of specifically binding with a model protein. In our model system, the functional group is 2,4-dinitrophenyl (DNP) and the protein is anti-DNP IgE (Immunoglobulin E). The functional polymer, &alpha;,&omega;-bi[2,4-dinitrophenyl caproic][poly(ethylene oxide)-b-poly(2-methoxystyrene)-b-poly(ethylene oxide)] (CDNP-PEO-P2MS-PEO-CDNP), is prepared by anionic living polymerization. The difunctional initiator utilized in the polymerization was prepared by electron transfer reaction of &alpha;-methylstyrene and potassium (mirror) metal. The 2-methoxystyrene monomer was added first to the initiator, followed by the addition of the second monomer, ethylene oxide, and finally the living polymer was terminated by methanol. The &alpha;,&omega;-dihydroxyl polymer [HO-PEO-P2MS-PEO-OH] was reacted with N-2,4-DNP-&isin;-amino caproic acid, by DCC coupling, resulting in the formation of &alpha;,&omega;-bi[2,4-dinitrophenylcaproic][poly(ethyleneoxide)-b-poly(2-methoxystyrene)-b-poly(ethylene oxide)] (CDNP-PEO-P2MS-PEO-CDNP). The polymers were characterized by FT-IR, 1H NMR and Gel Permeation Chromatography (GPC). The molecular weight distributions of the polymers were narrow (1.1-1.2) and polymers with molecular weights greater than 50,000 was used in this study. The polymers were yellow powders and soluble in tetrahydrofuran. A water soluble CDNP-PEO-P2MS-PEO-CDNP/ DMEG (dimethoxyethylene glycol) complex binds and achieves steady state binding with solution IgE within a few seconds. Higher molecular weight (water insoluble i.e. around 50,000) CDNP-PEO-P2MS-PEO-CDNP polymers, containing 1% single wall carbon nanotubes (SWCNT) were processed into electroactive nanofibers (100 nm to 500 nm in diameter) on silicon substrate. Fluorescence spectroscopy shows that anti-DNP IgE interacts with the nanofibers by binding with the DNP functional groups decorating the fibers. These observations suggest that appropriately functionalized nanofibers hold promise for developing biomarker detection device.

An efficient approach for preparing nanofibers decorated with functional groups capable of specifically interacting with proteins is described. The approach first requires the preparation of a polymer functionalized with the appropriate functional group. The functional polymer is fabricated into nanofibers by electrospinning. The effectiveness of the binding of the nanofibers with a protein is studied by confocal microscopy.

制作生物功能纳米纤维的过程

Electrospinning is an effective processing method for preparing nanofibers decorated with functional groups. Nanofibers decorated with functional groups ...

生物学

使用碳纳米纤维阵列将生物分子和染料高效递送到植物中

Using Vertically Aligned Carbon Nanofiber Arrays on Rigid or Flexible Substrates for Delivery of Biomolecules and Dyes to Plants

The delivery of biomolecules and impermeable dyes to intact plants is a major challenge. Nanomaterials are up-and-coming tools for the delivery of DNA to plants. As exciting as these new tools are, they have yet to be widely applied. Nanomaterials fabricated on rigid substrate (backing) are particularly difficult to successfully apply to curved plant structures. This study describes the process for microfabricating vertically aligned carbon nanofiber arrays and transferring them from a rigid to a flexible substrate. We detail and demonstrate how these fibers (on either rigid or flexible substrates) can be used for transient transformation or dye (e.g., fluorescein) delivery to plants. We show how VACNFs can be transferred from rigid silicon substrate to a flexible SU-8 epoxy substrate to form flexible VACNF arrays. To overcome the hydrophobic nature of SU-8, fibers in the flexible film were coated with a thin silicon oxide layer (2-3 nm). To use these fibers for delivery to curved plant organs, we deposit a 1 &#181;L droplet of dye or DNA solution on the fiber side of VACNF films, wait 10 min, place the films on the plant organ and employ a swab with a rolling motion to drive fibers into plant cells. With this method, we have achieved dye and DNA delivery in plant organs with curved surfaces.

作者聚焦：通过使用碳纳米纤维阵列进行创新来解锁工厂转型 

Here we describe methods for microfabricating vertically aligned carbon nanofibers (VACNFs), transferring VACNFs to flexible substrates, and applying VACNFs on both rigid and flexible substrates to plants for biomolecule and dye delivery.

在刚性或柔性基材上使用垂直排列的碳纳米纤维阵列，将生物分子和染料输送到植物

The delivery of biomolecules and impermeable dyes to intact plants is a major challenge. Nanomaterials are up-and-coming tools for the delivery of DNA to ...