这种材料是由美国能源部能源，材料科学和工程处（奖号DEFG - 02 - 07ER46471）和美国能源部能源研究中心光材料，在能源转换交互作用（支持工作的基础上奖“号DE - SC0001293 ），并从材料微分析中心内的弗雷德里克塞茨材料研究实验室（FSMRL）受惠。

1。简介<ol><li>本文演示的1D，2D和3D印刷导电微电极通过直接编写汇编。 </li><li>直接写入大会是建立集中墨水通过细喷嘴沉积1D - 3D印刷结构的方法。 </li><li>我们的系统由计算机控制的3轴平移台，墨水和喷嘴，以及10倍伸缩式镜头成像（图1）。 </li><li>直接写入大会是丝状的印刷方法，集中油墨挤压通过圆柱型喷嘴，其直径范围从0.1到250微米（图2）。值得注意的是，由于粘弹性油墨的特点，直接编写汇编使自收自支跨越功能（图3）。迄今为止，油墨的种类繁多，包括陶瓷 6,7组成的有机 8-10 11-15金属，聚合物16,17，18,19溶胶-凝胶材料已开发这种印刷方式（图4）。 </li></ol> 2。高度集中的银纳米粒子墨水的制备<ol><li>银纳米粒子墨水的准备，首先溶解在50克水和40克乙醇胺（2.1视频）的混合物5000的融合和聚（丙烯酸50000）分子量。 </li><li>聚合物行为作为覆盖剂控制的银纳米粒子的大小。 </li><li>下一步，硝酸银水溶液注入聚合物溶液。此外，后一个淡黄色透明溶液（2.2视频）。 </li><li>搅拌在室温下24小时后，该解决方案，开发了红褐色（2.3视频），恰逢与银纳米粒子的直径为5 nm，透射电子显微镜确定的形成。 </li><li>接下来，该解决方案是超声在65℃水浴为进一步粒子增长（2.4视频）2小时。 </li><li> Afteř超声，解决的办法是转移到500毫升烧杯中，冷却至室温。然后，300毫升乙醇，30毫升/分钟的速度滴定。由于乙醇是一个聚（丙烯酸）封盖剂的不良溶剂，颗粒迅速凝聚，并从沉淀的解决方案（视频2.5）。 </li><li>调迁上清液后，沉淀收集到离心管，离心20分钟9000 RPM（2.6视频）。 </li><li>这一步后，获得一个加载〜85％的固体与高度集中的银纳米粒子墨水（视频2.7）。 </li><li>对油墨的粘度和弹性模量的进一步控制，可以实现通过稀释，由同质化。举一个例子，一个保湿的解决方案，如乙二醇，可以添加墨水，然后在2000转速为3分钟，用一个Thinky同质化混频器匀浆。经过这个过程中，统一的一个蓝色洋红色墨水（视频2.8）。 </li><li> TEM图像显示银纳米粒子通过这个合成过程（图5_left）。粒子具有与5-50纳米大小分布的平均直径为20 nm。印刷结构的要求后退火，以提高其导电。不到30分钟后在250℃退火，银纳米粒子形成导电微电极电阻率接近10-5Ω•CM（图5_bottom权利）。退火温度的函数印刷银微电极的微观结构演变是在图5_top权利中所示。随着温度的升高，从150 °到550 ° C，微电极经过〜30％的总体积收缩致密化。11 </li><li>强烈取决于其固相含量，油墨的流变性，决定了它的印刷适性。油墨的粘度增加固相含量的增加（图6）。由于稀释油墨粘度低的结果在一个显著的横向扩散，concentr一个固体装载中的相应范围从70至85 wt％的油墨需要印刷的平面和跨越油墨细丝。 </li><li>油墨的弹性模量增加固相含量增加（图7）。线性粘弹区，弹性模量上升了近三个数量级，作为固​​相含量增加60至75 WT％。 2000帕弹性模量是一个最低要求出示自收自支或跨越功能。 </li></ol> 3。直接编写汇编<ol><li>直接写入大会是由先装入一个针筒墨水。附加沉积喷嘴后，墨水装入针筒安装到3轴印刷阶段（视频3.1）。 </li><li>使用一个计算机程序，任意设计，包括线性的，平面的，复杂的三维结构，可方便地生成（3.2视频）。 </li><li>接下来，喷嘴高度调整望远镜的镜头具有10倍变焦（视频3.3）的援助。 </l我&gt; <li>使用空气为动力的的流体点胶系统施加压力后，油墨沉积到基板与控制打印速度（3.4视频）。所需的压力取决于油墨的流变性，喷嘴直径，印刷速度，但典型值范围在20-500微米/秒，从10-100 PSI这种印刷是在室温空气中。使用这种印刷过程，在不同的布局和大小规模的银微电极印刷证明。 </li><li>举一个例子，中心到中心线间距为100μm的导电银网格，由一个硅晶片衬底上的5微米的喷嘴图案的印刷演示（视频3.5）。 </li><li>此外，该视频展示了如何创建一个高宽比圆柱形结构由30微米的喷嘴使用层层印刷方法（视频3.6）。 </li><li>此外，两块玻璃基板之间的银微电极的全方位印刷偏移1毫米的高度DIfference证明，使用30微米的喷嘴（视频3.7）。 </li><li>完全独立的，垂直印银microspikes可以创建由一个硅晶片的基板（视频3.8）的30微米的喷嘴。 </li><li>最后，这个视频显示了使用10微米的喷嘴（视频3.9）跨越银电极直接书写。打印功能可以跨越距离可达一厘米，最小的下垂或屈曲。 </li></ol> 4。代表性的成果： 我们准备了一个高度集中的银纳米粒子墨水和证明的打印分辨率为1〜2的电子和光电应用的平面和三维图案印在导电功能 - 30微米。举一个例子，图8展示了这项技术的打印分辨率。 〜2微米（1.4微米厚）的最小电极宽度印刷的特点是在一个单一的通过使用1 微米的喷嘴11获得。 图9显示小号透明导电银网格，图案由5微米的喷嘴在一个灵活的聚酰亚胺薄膜 12 。印刷电网下面的文本都清晰可见。这些透明的银色网格可以为透明导电氧化物（TCO）材料吸引力的替代品。 到非平面基板的共形印花也用这种方法启用。图10展示了一个三维电小天线的形印刷。一个100微米的金属喷嘴，用来打印在玻璃表面半球13蜿蜒的线条图案。这种方法可能会发现几个应用程序，包括植入式和可穿戴式天线，电子，传感器。 跨越银微电极在三维光伏和发光二极管的应用演示（图11-14）。 首先，图11是硅球壳的一个例子。这与2微米吨的脆弱电影hickness可以引线键合到一个外部电路的全方位印刷14。此方法使用最小的接触压力，这是非常微妙的设备优势。 下一步，图12演示了一个例子打印一个硅太阳能电池硅microribbon元素是由33微米的差距15分离的微蜂窝阵列的跨越互连。 下一步，图13显示了银互联砷化镓为基础的LED阵列为4 × 4像素，其中每个像素（500 × 500 × 2.5微米3）间距为200微米， 除了 11 。底部图像显示的LED阵列，从单个像素的6 V偏压下的统一的红光发光。跨越电极的打印能力，能够在不支持或牺牲层使用（顶端图像）多层互连。 作为最后的示范，图14显示了复杂的三维MIC的SEM图像印roperiodic银格由5μm的喷嘴。 <img alt="图1" src="/files/ftp_upload/3189/3189fig1.jpg" /> 图1直接墨水书写器具的光学图像。 <img alt="图2" src="/files/ftp_upload/3189/3189fig2.jpg" /> 图2。直接墨水写了丝状功能。 <img alt="图3" src="/files/ftp_upload/3189/3189fig3.jpg" /> 图3。自收自支跨越功能的直接的墨水书写。 <img alt="图4" src="/files/ftp_upload/3189/3189fig4.jpg" /> 图4。直接墨水书写的墨水设计。一个广泛集中粘弹性油墨已经开发平面和复杂的三维结构与微观特征的直写。 <img alt="图5" src="/files/ftp_upload/3189/3189fig5.jpg" /> 图5（左）透射电子显微镜（TEM）银纳米粒子的形象。 （右上）与退火温度的功能，作为一个15微米的喷嘴图案的银微电极的SEM图像。 （右下）电器的退火温度和时间的功能的银微电极的电阻率。 <img alt="图6" src="/files/ftp_upload/3189/3189fig6.jpg" /> 图6。表观粘度的固体加载功能的银纳米粒子墨水（η） 。 <img alt="图7" src="/files/ftp_upload/3189/3189fig7.jpg" /> 图7。剪切弹性模量（G&#39;）作为不同固相含量的银纳米粒子墨水功能的剪应力。 <img alt="图8" src="/files/ftp_upload/3189/3189fig8.jpg" /> 图8。SEM图像的平面图案的银微电极阵列1微米的喷嘴的硅晶片上。 <img alt="图9" src="/files/ftp_upload/3189/3189fig9.jpg" /> 图9。光学图像的透明导电银网格（左）和SEM图像印刷的网格线间距（右）的功能。 <img alt="图10" src="/files/ftp_upload/3189/3189fig10.jpg" /> 图10：在一个半球形玻璃基板上的电小天线的形印刷拍摄的光学图像。 <img alt="图11" src="/files/ftp_upload/3189/3189fig11.jpg" /> 图11到薄（2微米）的硅球壳跨越银微电极印刷过程中获得的光学图像。 <img alt="图12" src="/files/ftp_upload/3189/3189fig12.jpg" /> 跨越的银微电极的SEM图像印到一个硅所以图 12。LAR微蜂窝阵列。 <img alt="图13" src="/files/ftp_upload/3189/3189fig13.jpg" /> 图13。SEM照片（上）和4 - 4 LED芯片的银微电极互连阵列的光学图像（底部） 。 <img alt="图14" src="/files/ftp_upload/3189/3189fig14.jpg" /> 图14扫描电镜图像的三维microperiodic银格。

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Mater. 21, 47-54 (2011).</li><li>Wu, W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Direct-write+assembly+of+biomimetic+microvascular+networks+for+efficient+fluid+transport.">Direct-write assembly of biomimetic microvascular networks for efficient fluid transport.</a> Soft. Matter. 6, 739-742 (2010).</li><li>Wu, W., DeConinck, A., Lewis, J. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Omnidirectional+Printing+of+3D+Microvascular+Networks.">Omnidirectional Printing of 3D Microvascular Networks.</a> Adv. Mater. 23, H178-H183 (2011).</li></ol>

没有利益冲突的声明。

基于液滴的传统的印刷方法，如喷墨打印，是有限的低长宽比由于稀的性质和所使用的油墨低粘度的平面电极的制造。近日，蘸笔纳米光刻技术（DPN）20-22和23-25 ​​E -喷气印刷已经用于导电功能模式。这条路线，还聘请稀，粘度低油墨。 Pearton和同事使用DPN的存款高达1600微米s - 1和线宽度约0.5微米22写入速度市售的银纳米粒子墨水。然而，制造大面积的重复性模式尚未证明这种方法。纳米银油墨也已存入电子喷射印花与〜1.5微米25线宽形成导电的痕迹。但是，喷墨打印，可能会出现不均匀的印刷特性，由于卫星液滴的形成和非均匀下降ðrying 24,25。 如上所述，集中的银纳米粒子墨水直接写大会通过丝状的印刷方法克服了这些局限性。这项技术使一个高宽比（H / W≈1.0）在允许一个单一的1D，2D和3D架构创造通过导电微电极的制作。印刷功能的大小取决于喷嘴直径，油墨固体装载，施加的压力，和印刷速度。迄今为止，小的导电痕迹〜2微米已使用温和的速度（&lt;2毫米S -1）1微米的喷嘴图案。通过调整油墨的组成和喷嘴的几何形状，最高印刷速度超过10厘米S -1是可能的。然而，使用精细的喷嘴（&lt;5微米）的高速印刷仍然是一个重大的挑战。 为了证明直接写入大会的应用，我们捏造导电电网，ELectrically小天线，太阳能电池，发光二极管，平面和跨越印刷电极（图8-14）。值得一提的是，我们的做法是没有限制的金属结构创造。如那些基于丝素蛋白，凝胶和逃犯的有机油墨，使用其他墨水设计，我们已经构建了3D支架和微血管网络，通过直接写入大会26日至 30日组织工程和细胞培养。 展望未来，有很多的机遇和挑战。进一步的进展需要新的墨水设计，更好的油墨血流动力学建模，和增强机器人和控制系统。 1D高吞吐量和纳米分辨率的三维结构的大面积制造（&lt;100 nm的）仍然是一个重大的挑战。

<table border="1"><tr><td> 试剂名称 </td><td> 公司 </td><td> 目录编号 </td><td> 评论 </td></tr><tr><td>聚（丙烯酸） </td><td> Polysciences，公司</td><td> 06519 </td><td>兆瓦5,000元克/摩尔</td></tr><tr><td>聚（丙烯酸） </td><td> Polysciences，公司</td><td> 00627 </td><td> MW 50,000克/摩尔</td></tr><tr><td>硝酸银</td><td> Sigma - Aldrich公司</td><td> 209139 </td><td>银源</td></tr><tr><td>二乙醇胺</td><td> Sigma - Aldrich公司</td><td> D8885 </td><td>溶剂/还原剂</td></tr><tr><td>乙二醇</td><td> Sigma - Aldrich公司</td><td> 102466 </td><td>保湿</td></tr><tr><td> Sonicater </td><td> Fisher Scientific则</td><td> FS30H </td><td> - </td></tr><tr><td>离心分离</td><td> Beckman Coulter公司</td><td>阿凡提商标的J - 25我</td><td> - </td></tr><tr><td>机器人阶段</td><td>艾特航空股份有限公司</td><td> ABL的900010 </td><td> 3轴运动</td></tr><tr><td>针筒</td><td> EFD公司</td><td> 5109LBP - B </td><td> 3毫升</td></tr><tr><td>喷嘴</td><td> EFD公司</td><td> - </td><td> ID = 0.1 - 250微米</td></tr><tr><td>饮水机</td><td> EFD公司</td><td> 800 </td><td>空气动力</td></tr><tr><td>设计软件</td><td>定制设计</td><td> - </td><td>明杰许</td></tr></table>

planar and three-dimensional printing of conductive inks

印刷电子产品依靠低成本，大面积的制造路线，以创建灵活或多维电子，光电， 和 1-3生物医学设备。在本文中，我们专注于一（1D），两（2D）和三维（3D）的形式灵活，伸展，并跨越微电极的导电金属油墨印刷。 直接写入大会4,5是一个1到3D印刷技术，使制造范围由沉积集中墨水通过喷嘴（〜0.1 - 250微米）的罚款 ​​，从简单的线条，结构复杂的特点。这种印刷方法由计算机控制的3轴的翻译阶段，墨水和喷嘴，和10倍伸缩式镜头的可视化。与喷墨打印，基于液滴的过程中，直接写入大会涉及挤压油墨细丝，或缩小的平面。印细丝通常符合喷嘴尺寸。 HencE，微观特性（&lt;1微米），可仿造和组装成更大的数组和多维架构。 在本文中，我们首先通过直接写装配平面和3D印刷合成一个高度集中的银纳米粒子墨水。其次，表现在多维图案印刷微电极的标准协议。最后，印刷微电极电小天线，太阳能电池，发光二极管的应用突出。

Watch this Scientific Journal Video about Planar and Three-Dimensional Printing of Conductive Inks at JoVE.com

Planar and Three-Dimensional Printing of Conductive Inks

导电金属油墨印刷平面和立体的描述。我们的方法制造印刷电子，光电，并在不寻常的布局在微尺度生物医学设备提供新的途径。

平面和立体印刷的导电油墨

Printed electronics rely on low-cost, large-area fabrication routes to create flexible or multidimensional electronic, optoelectronic, and biomedical ...

planar-and-three-dimensional-printing-of-conductive-inks

Research

JoVE Journal

Bioengineering

36.9K 次观看.  University of Illinois at Urbana-Champaign. 导电金属油墨印刷平面和立体的描述。我们的方法制造印刷电子，光电，并在不寻常的布局在微尺度生物医学设备提供新的途径。

视频: Planar and Three-Dimensional Printing of Conductive Inks

Three-dimensional Optical-resolution Photoacoustic Microscopy

Optical microscopy, providing valuable insights at the cellular and organelle levels, has been widely recognized as an enabling biomedical technology. As the mainstays of in vivo three-dimensional (3-D) optical microscopy, single-/multi-photon fluorescence microscopy and optical coherence tomography (OCT) have demonstrated their extraordinary sensitivities to fluorescence and optical scattering contrasts, respectively. However, the optical absorption contrast of biological tissues, which encodes essential physiological/pathological information, has not yet been assessable. 
The emergence of biomedical photoacoustics has led to a new branch of optical microscopy optical-resolution photoacoustic microscopy (OR-PAM)1, where the optical irradiation is focused to the diffraction limit to achieve cellular1 or even subcellular2 level lateral resolution. As a valuable complement to existing optical microscopy technologies, OR-PAM brings in at least two novelties. First and most importantly, OR-PAM detects optical absorption contrasts with extraordinary sensitivity (i.e., 100%). Combining OR-PAM with fluorescence microscopy3 or with optical-scattering-based OCT4 (or with both) provides comprehensive optical properties of biological tissues. Second, OR-PAM encodes optical absorption into acoustic waves, in contrast to the pure optical processes in fluorescence microscopy and OCT, and provides background-free detection. The acoustic detection in OR-PAM mitigates the impacts of optical scattering on signal degradation and naturally eliminates possible interferences (i.e., crosstalks) between excitation and detection, which is a common problem in fluorescence microscopy due to the overlap between the excitation and fluorescence spectra. 
Unique for optical absorption imaging, OR-PAM has demonstrated broad biomedical applications since its invention, including, but not limited to, neurology5, 6, ophthalmology7, 8, vascular biology9, and dermatology10. In this video, we teach the system configuration and alignment of OR-PAM as well as the experimental procedures for in vivo functional microvascular imaging.

Optical-resolution photoacoustic microscopy (OR-PAM) is an emerging technology capable of imaging optical absorption contrasts in vivo with cellular resolution and sensitivity. Here, we provide a visualized instruction on the experimental protocols of OR-PAM, including system configuration, system alignment, typical in vivo experimental procedures, and functional imaging schemes.

三维光学分辨率的光声显微镜

Optical microscopy, providing valuable insights at the cellular and organelle levels, has been widely recognized as an enabling biomedical technology. As ...

科研

生物工程

Alginate Hydrogels for Three-Dimensional Organ Culture of Ovaries and Oviducts

Ovarian cancer is the fifth leading cause of cancer deaths in women and has a 63% mortality rate in the United States1. The cell type of origin for ovarian cancers is still in question and might be either the ovarian surface epithelium (OSE) or the distal epithelium of the fallopian tube fimbriae2,3. Culturing the normal cells as a primary culture in vitro will enable scientists to model specific changes that might lead to ovarian cancer in the distinct epithelium, thereby definitively determining the cell type of origin. This will allow development of more accurate biomarkers, animal models with tissue-specific gene changes, and better prevention strategies targeted to this disease.
Maintaining normal cells in alginate hydrogels promotes short term in vitro culture of cells in their three-dimensional context and permits introduction of plasmid DNA, siRNA, and small molecules. By culturing organs in pieces that are derived from strategic cuts using a scalpel, several cultures from a single organ can be generated, increasing the number of experiments from a single animal. These cuts model aspects of ovulation leading to proliferation of the OSE, which is associated with ovarian cancer formation. Cell types such as the OSE that do not grow well on plastic surfaces can be cultured using this method and facilitate investigation into normal cellular processes or the earliest events in cancer formation4.
Alginate hydrogels can be used to support the growth of many types of tissues5. Alginate is a linear polysaccharide composed of repeating units of &beta;-D-mannuronic acid and &alpha;-L-guluronic acid that can be crosslinked with calcium ions, resulting in a gentle gelling action that does not damage tissues6,7. Like other three-dimensional cell culture matrices such as Matrigel, alginate provides mechanical support for tissues; however, proteins are not reactive with the alginate matrix, and therefore alginate functions as a synthetic extracellular matrix that does not initiate cell signaling5. The alginate hydrogel floats in standard cell culture medium and supports the architecture of the tissue growth in vitro.
A method is presented for the preparation, separation, and embedding of ovarian and oviductal organ pieces into alginate hydrogels, which can be maintained in culture for up to two weeks. The enzymatic release of cells for analysis of proteins and RNA samples from the organ culture is also described. Finally, the growth of primary cell types is possible without genetic immortalization from mice and permits investigators to use knockout and transgenic mice.

Culture of normal cells in their three-dimensional context represents an alternative method to study early events required for cellular transformation and tumorigenesis. This method is used to grow normal ovarian and oviductal cells to study early events in ovarian cancer formation.

卵巢和输卵管的三维器官培养的海藻酸钙凝胶

Ovarian cancer is the fifth leading cause of cancer deaths in women and has a 63% mortality rate in the United States1. The cell type of origin for ...

Investigating the Three-dimensional Flow Separation Induced by a Model Vocal Fold Polyp

The fluid-structure energy exchange process for normal speech has been studied extensively, but it is not well understood for pathological conditions. Polyps and nodules, which are geometric abnormalities that form on the medial surface of the vocal folds, can disrupt vocal fold dynamics and thus can have devastating consequences on a patient&#39;s ability to communicate. Our laboratory has reported particle image velocimetry (PIV) measurements, within an investigation of a model polyp located on the medial surface of an in&#160;vitro driven vocal fold model, which show that such a geometric abnormality considerably disrupts the glottal jet behavior. This flow field adjustment is a likely reason for the severe degradation of the vocal quality in patients with polyps. A more complete understanding of the formation and propagation of vortical structures from a geometric protuberance, such as a vocal fold polyp, and the resulting influence on the aerodynamic loadings that drive the vocal fold dynamics, is necessary for advancing the treatment of this pathological condition. The present investigation concerns the three-dimensional flow separation induced by a wall-mounted prolate hemispheroid with a 2:1 aspect ratio in cross flow, i.e. a model vocal fold polyp, using an oil-film visualization technique. Unsteady, three-dimensional flow separation and its impact of the wall pressure loading are examined using skin friction line visualization and wall pressure measurements.

Vocal fold polyps can disrupt vocal fold dynamics and thus can have devastating consequences on a patient&#39;s ability to communicate. Three-dimensional flow separation induced by a wall-mounted model polyp and its impact on the wall pressure loading are examined using particle image velocimetry, skin friction line visualization, and wall pressure measurements.

研究模型声乐折叠聚合物引起的三维流量分离

The fluid-structure energy exchange process for normal speech has been studied extensively, but it is not well understood for pathological conditions. ...

Human Cartilage Tissue Fabrication Using Three-dimensional Inkjet Printing Technology

Bioprinting, which is based on thermal inkjet printing, is one of the most attractive enabling technologies in the field of tissue engineering and regenerative medicine. With digital control&#160;cells, scaffolds, and growth factors can be precisely deposited to the desired two-dimensional (2D) and three-dimensional (3D) locations rapidly. Therefore, this technology is an ideal approach to fabricate tissues mimicking their native anatomic structures. In order to engineer cartilage with native zonal organization, extracellular matrix composition (ECM), and mechanical properties, we developed a bioprinting platform using a commercial inkjet printer with simultaneous photopolymerization capable for 3D cartilage tissue engineering. Human chondrocytes suspended in poly(ethylene glycol) diacrylate (PEGDA) were printed for 3D neocartilage construction via layer-by-layer assembly. The printed cells were fixed at their original deposited positions, supported by the surrounding scaffold in simultaneous photopolymerization. The mechanical properties of the printed tissue were similar to the native cartilage. Compared to conventional tissue fabrication, which requires longer UV exposure, the viability of the printed cells with simultaneous photopolymerization was significantly higher. Printed neocartilage demonstrated excellent glycosaminoglycan (GAG) and collagen type II production, which was consistent with gene expression. Therefore, this platform is ideal for accurate cell distribution and arrangement for anatomic tissue engineering.

The methods described in this paper show how to convert a commercial inkjet printer into a bioprinter with simultaneous UV polymerization. The printer is capable of constructing 3D tissue structure with cells and biomaterials. The study demonstrated here constructed a 3D neocartilage.

人体软骨组织制作采用三维喷墨打印技术

Bioprinting, which is based on thermal inkjet printing, is one of the most attractive enabling technologies in the field of tissue engineering and ...

Analysis of Cell Migration within a Three-dimensional Collagen Matrix

The ability to migrate is a hallmark of various cell types and plays a crucial role in several physiological processes, including&#160;embryonic development, wound healing, and immune responses. However, cell migration is also a key mechanism in cancer enabling these cancer cells to detach from the primary tumor to start metastatic spreading. Within the past years various cell migration assays have been developed to analyze the migratory behavior of different cell types. Because the locomotory behavior of cells markedly differs between a two-dimensional (2D) and three-dimensional (3D) environment it can be assumed that the analysis of the migration of cells that are embedded within a 3D environment would yield in more significant cell migration data. The advantage of the described 3D collagen matrix migration assay is that cells are embedded within a physiological 3D network of collagen fibers representing the major component of the extracellular matrix. Due to time-lapse video microscopy real cell migration is measured allowing the determination of several migration parameters as well as their alterations in response to pro-migratory factors or inhibitors. Various cell types could be analyzed using this technique, including lymphocytes/leukocytes, stem cells, and tumor cells. Likewise, also cell clusters or spheroids could be embedded within the collagen matrix concomitant with analysis of the emigration of single cells from the cell cluster/ spheroid into the collagen lattice. We conclude that the 3D collagen matrix migration assay is a versatile method to analyze the migration of cells within a physiological-like 3D environment.

Cell migration is a biological phenomenon that is involved in a plethora of physiological, such as wound healing and immune responses, and pathophysiological processes, like cancer. The 3D-collagen matrix migration assay is a versatile tool to analyze the migratory properties of different cell types within in a 3D physiological-like environment.

在一个三维胶原蛋白矩阵分析细胞迁移

The ability to migrate is a hallmark of various cell types and plays a crucial role in several physiological processes, including&#160;embryonic ...

Generation of a Three-dimensional Full Thickness Skin Equivalent and Automated Wounding

In vitro models are a cost effective and ethical alternative to study cutaneous wound healing processes. Moreover, by using human cells, these models reflect the human wound situation better than animal models. Although two-dimensional models are widely used to investigate processes such as cellular migration and proliferation, models that are more complex are required to gain a deeper knowledge about wound healing. Besides a suitable model system, the generation of precise and reproducible wounds is crucial to ensure comparable results between different test runs. In this study, the generation of a three-dimensional full thickness skin equivalent to study wound healing is shown. The dermal part of the models is comprised of human dermal fibroblast embedded in a rat-tail collagen type I hydrogel. Following the inoculation with human epidermal keratinocytes and consequent culture at the air-liquid interface, a multilayered epidermis is formed on top of the models. To study the wound healing process, we additionally developed an automated wounding device, which generates standardized wounds in a sterile atmosphere.

The goal of this protocol is to build up a three-dimensional full thickness skin equivalent, which resembles natural skin. With a specifically constructed automated wounding device, precise and reproducible wounds can be generated under maintenance of sterility.

三维全层皮肤的生成等效和自动伤人

In vitro models are a cost effective and ethical alternative to study cutaneous wound healing processes. Moreover, by using human cells, these models ...

Preparation and Analysis of In Vitro Three Dimensional Breast Carcinoma Surrogates

Three dimensional (3D) culture is a more physiologically relevant method to model cell behavior in vitro than two dimensional culture. Carcinomas, including breast carcinomas, are complex 3D tissues composed of cancer epithelial cells and stromal components, including fibroblasts and extracellular matrix (ECM). Yet most in vitro models of breast carcinoma consist only of cancer epithelial cells, omitting the stroma and, therefore, the 3D architecture of a tumor in vivo. Appropriate 3D modeling of carcinoma is important for accurate understanding of tumor biology, behavior, and response to therapy. However, the duration of culture and volume of 3D models is limited by the availability of oxygen and nutrients within the culture. Herein, we demonstrate a method in which breast carcinoma epithelial cells and stromal fibroblasts are incorporated into ECM to generate a 3D breast cancer surrogate that includes stroma and can be cultured as a solid 3D structure or by using a perfusion bioreactor system to deliver oxygen and nutrients. Following setup and an initial growth period, surrogates can be used for preclinical drug testing. Alternatively, the cellular and matrix components of the surrogate can be modified to address a variety of biological questions. After culture, surrogates are fixed and processed to paraffin, in a manner similar to the handling of clinical breast carcinoma specimens, for evaluation of parameters of interest. The evaluation of one such parameter, the density of cells present, is explained, where ImageJ and CellProfiler image analysis software systems are applied to photomicrographs of histologic sections of surrogates to quantify the number of nucleated cells per area. This can be used as an indicator of the change in cell number over time or the change in cell number resulting from varying growth conditions and treatments.

Preparation and Analysis of In Vitro Three Dimensional Breast Carcinoma Surrogates

We demonstrate a method to generate 3D breast cancer surrogates, which can be cultured using a perfusion bioreactor system to deliver oxygen and nutrients. Following growth, surrogates are fixed and processed to paraffin for evaluation of parameters of interest. The evaluation of one such parameter, cell density, is explained.

制备及分析<em&gt;体外</em&gt;三维乳腺癌代理项

Three dimensional (3D) culture is a more physiologically relevant method to model cell behavior in vitro than two dimensional culture. Carcinomas, ...

Engineering Three-dimensional Epithelial Tissues Embedded within Extracellular Matrix

The architecture of branched organs such as the lungs, kidneys, and mammary glands arises through the developmental process of branching morphogenesis, which is regulated by a variety of soluble and physical signals in the microenvironment. Described here is a method created to study the process of branching morphogenesis by forming engineered three-dimensional (3D) epithelial tissues of defined shape and size that are completely embedded within an extracellular matrix (ECM). This method enables the formation of arrays of identical tissues and enables the control of a variety of environmental factors, including tissue geometry, spacing, and ECM composition. This method can also be combined with widely used techniques such as traction force microscopy (TFM) to gain more information about the interactions between cells and their surrounding ECM. The protocol can be used to investigate a variety of cell and tissue processes beyond branching morphogenesis, including cancer invasion.

This manuscript describes a soft lithography-based technique to engineer uniform arrays of three-dimensional (3D) epithelial tissues of defined geometry surrounded by extracellular matrix. This method is amenable to a wide variety of cell types and experimental contexts and allows for high-throughput screening of identical replicates.

工程三维上皮组织细胞外基质中嵌入

The architecture of branched organs such as the lungs, kidneys, and mammary glands arises through the developmental process of branching morphogenesis, ...

Fabrication of Three-dimensional Paper-based Microfluidic Devices for Immunoassays

Paper wicks fluids autonomously due to capillary action. By patterning paper with hydrophobic barriers, the transport of fluids can be controlled and directed within a layer of paper. Moreover, stacking multiple layers of patterned paper creates sophisticated three-dimensional microfluidic networks that can support the development of analytical and bioanalytical assays. Paper-based microfluidic devices are inexpensive, portable, easy to use, and require no external equipment to operate. As a result, they hold great promise as a platform for point-of-care diagnostics. In order to properly evaluate the utility and analytical performance of paper-based devices, suitable methods must be developed to ensure their manufacture is reproducible and at a scale that is appropriate for laboratory settings. In this manuscript, a method to fabricate a general device architecture that can be used for paper-based immunoassays is described. We use a form of additive manufacturing (multi-layer lamination) to prepare devices that comprise multiple layers of patterned paper and patterned adhesive. In addition to demonstrating the proper use of these three-dimensional paper-based microfluidic devices with an immunoassay for human chorionic gonadotropin (hCG), errors in the manufacturing process that may result in device failures are discussed. We expect this approach to manufacturing paper-based devices will find broad utility in the development of analytical applications designed specifically for limited-resource settings.

We detail a method to fabricate three-dimensional paper-based microfluidic devices for use in the development of immunoassays. Our approach to device assembly is a type of multilayer, additive manufacturing. We demonstrate a sandwich immunoassay to provide representative results for these types of paper-based devices.

三维纸基微流体装置的制造用于免疫分析

Paper wicks fluids autonomously due to capillary action. By patterning paper with hydrophobic barriers, the transport of fluids can be controlled and ...

Three-dimensional Printing of Thermoplastic Materials to Create Automated Syringe Pumps with Feedback Control for Microfluidic Applications

Microfluidics has become a critical tool in research across the biological, chemical, and physical sciences. One important component of microfluidic experimentation is a stable fluid handling system capable of accurately providing an inlet flow rate or inlet pressure. Here, we have developed a syringe pump system capable of controlling and regulating the inlet fluid pressure delivered to a microfluidic device. This system was designed using low-cost materials and additive manufacturing principles, leveraging three-dimensional (3D) printing of thermoplastic materials and off-the-shelf components whenever possible. This system is composed of three main components: a syringe pump, a pressure transducer, and a programmable microcontroller. Within this paper, we detail a set of protocols for fabricating, assembling, and programming this syringe pump system. Furthermore, we have included representative results that demonstrate high-fidelity, feedback control of inlet pressure using this system. We expect this protocol will allow researchers to fabricate low-cost syringe pump systems, lowering the entry barrier for the use of microfluidics in biomedical, chemical, and materials research.

Here we present a protocol to construct a pressure-controlled syringe pump to be used in microfluidic applications. This syringe pump is made from an&#160;additively manufactured body, off-the-shelf hardware, and open-source electronics. The resulting system is low-cost, straightforward to build, and delivers well-regulated fluid flow to enable rapid microfluidic research.

三维热塑性材料的印刷, 以创建自动注射器泵的反馈控制微流控应用

Microfluidics has become a critical tool in research across the biological, chemical, and physical sciences. One important component of microfluidic ...