这项工作是由赠款的援助从教育部，文化，体育，日本的科学技术，京都技术科学中心，和三谷的基础研究与发展青年科学家（16K17956）的支持。

1.仪器<ol><li> 偏振元件 <ol><li>为了构建一个偏振装置，提供具有卤素灯，导光，偏振器，一个样品台，光学轨道，杆架，和一个数字单镜头反光照相机（光源参见偏振图1C和材料清单装置部分）。 </li></ol></li></ol> 2.生物聚合物溶液的制备<ol><li> 多糖溶液 <ol><li>通过在〜80℃下搅拌超过12小时溶解SACRAN 7（0.5g）在纯水（100mL）中。在溶解，用保鲜膜覆盖，以防止蒸发容器。制备的黄原胶以同样的方式的水溶液。 </li><li>在〜25℃下冷却该溶液，以获得0.5重量％的水溶液。 </li><li>离心SACRAN溶液以除去杂质（48400×g下，4℃，1个小时，3添ES）。 </li></ol></li><li> MT解决方案 <ol><li>在的Britton-Robinson缓冲液制备0.5重量％的微管蛋白溶液（1毫升）（80mM的哌嗪N，N&#39; -双（2-乙磺酸）（PIPES）; 1mM的乙二醇-双（β氨基乙醚） - N，N，N &#39;N&#39;，N&#39;-四乙酸（EGTA）;在冰上8和5mM MgCl 2的，pH为6.8）。 </li><li>使用0.5重量％的微管蛋白溶液（50μL）和鸟苷-5&#39; - [（α，β）-methyleno]三磷酸（GpCpp）（5μL），制备含GpCpp微管蛋白溶液（50微升）。孵育在37℃下3小时，以获得稳定的核MT。 注:GpCpp的作用是支持MT形成和完全抑制MT的解聚微管蛋白。 </li><li>混合在约25℃的0.5重量％的微管蛋白溶液（950μL）和含GpCpp微管蛋白溶液（50μL）1天，以获得一个稳定的0.5重量％溶液MT。 </li></ol></li><li> DNA溶液<ol><li>准备在Tris-EDTA缓冲溶液中的0.5重量％的DNA溶液（1毫升）（10毫摩尔Tris，pH8.0中，用1mM EDTA）中。 </li></ol></li><li>保持0.5重量％的生物聚合物的样品溶液在25℃下为干燥实验。 </li></ol> 3，干燥下交叉偏振光实验和观察<ol><li> 在单侧打开的细胞溶液（图1A） <ol><li>切断的硅片（参见材料列表）成厚度为1mm的适当形状（5-15毫米1毫米内部尺寸，以及约20毫米; 图1A）。 <ol><li>组装用的5-15毫米1毫米内尺寸的硅隔离物构成的单侧开放单元，和约20毫米和两个非修饰的载玻片（76 1mm×1mm的×26毫米）。修正了双剪辑池的两边事先保留样品溶液泄漏。 </li></ol></li><li>慢慢添加的每个使用〜1毫米孔径的移液管的0.5重量％的生物聚合物溶液（100-300μL）的TE尖端给每个小区在〜25℃。使用注射器针头将细胞去除气泡。 </li><li>放置细胞与在60℃下用于蒸发大气压力下在空气循环的烘箱蒸镀方向是相反的重力。 </li></ol></li><li> 下交叉偏振光观测（图1B-1C） <ol><li>通过具有平坦表面光源在很宽的区域（80毫米×80mm）中一个100瓦的卤素灯提供直的可见光。调整偏振器使用保持器（ 图1C）在45°和135°。 </li><li>固定使用光学轨道和杆支架（ 图1C）的光源，偏振器，样品台，和照相机的位置。放置在两个偏振器之间的样品台（偏振片之间的距离应为约5厘米）。将相机约20厘米的样品台上，以允许聚焦。 </li><li>在给定时间，将样品从步骤3.1.3 POLA之间上平行于XZ平面的阶段rizers并覆盖有黑色帘装置;实际装置被示于图1C中 。 </li><li>拍摄通过线性交叉的偏振器使用的数字单镜头反射式照相机用标准变焦镜头的样品（见材料列表）。控制照相机设置，例如焦距，使用计算机软件（参见材料列表）。 </li></ol></li><li> 透射光强度的空间-时间分析（图1C） <ol><li>为了评估在干燥过程中的取向序参数的变化，收集照片每小时24小时。 </li><li>测量沿着作为使用图像处理程序（ 例如，使用ImageJ）的灰度值在Z方向上的中心线的透射光强度。 </li><li>画出从上开口侧的距离的函数的灰度值的曲线图。 </li></ol></li><li> 交叉极化二极管灯下显微观察吨（图1D） <ol><li>要检查的方法中，使显微观察用配有CCD照相机9的偏光显微镜。保持一阶延迟板在光路中。控制使用PC软件的照片的条件。 </li></ol></li></ol>

<ol>
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作者什么都没有透露。

有时很难为摄像机聚焦在样品上，由于透射光强度太低。在这种情况下，将延长的透明塑料薄膜上的阶段帮助安排的焦点。可观察到的分辨率的限制是依赖于摄像机镜头上，〜10微米在这种情况下。样品厚度，ΔY，的可观察的限制是依赖于该灯的最大光强度，〜在这种情况下10mm以下。 在图1B所示的装置的优点是，它允许侧视图样品的照片，以在干燥过程中作出。观察可以不依靠在水平平面上，这是在典型的显微镜中使用来进行。通过干燥实验期间站在单侧开孔，蒸发方向被限制，在重力的相反方向。监测侧视图还使干燥的大鼠的计算E 4。 在未来，通过将温度和湿度调节器在所述装置的样品台上，将有可能自动地监视在向秩序参数随时间的变化。此外，在建立一个平衡监测体重变化，将有助于浓度的估计。此方法也可用于澄清各向异性溶胀过程的多糖水凝胶11，12。因此，将可能监视软致动器的结构变化。

通过聚焦于生物聚合物的刚性，棒状的结构，动态软材料已被用于各种应用，包括多糖生物膜基质1中，"活性凝胶"细胞骨架蛋白2构成，并且所希望的形状3的"DNA折纸"。为了阐明的结构特性，许多策略已探索，如透射电子显微镜，扫描电子显微镜，原子力显微镜，和共聚焦荧光显微镜。然而，因为这些方法都处于干燥状态或静止状态进行的大部分，这是很难解释的动态行为在宏观尺度，在实际生活系统看到。最近，我们成功地通过偏振光4中观察到的水气-LC界面上的生物聚合物的动态行为。在取向的结构的可视化，同时干燥该生物聚合物解决方案，随时间的变化表明不稳定的空气-LC接口上的生物聚合物的自我整合。 在这里，我们描述了用于LC的生物聚合物溶液在使用偏振仪器空气-LC界面处的干燥的协议。而不是在LC相的其他分析不考虑干燥5， 图6，在干燥过程中的LC动力学通过在一个单侧开放单元评价在流体相中的侧视图的取向顺序参数这里调查。单元蒸发的组合和使用允许以受控的蒸发方向宏观监测偏振仪器。此外，有可能通过集中在吸附微区，将其受分子量，浓度等的晶体结构为了证明该方法的有效性来验证干燥记录，干燥亲与刚性杆形状基本生物聚合物，如多糖，微管（MTS），和DNA的正如事实，进行了调查。我们选择这些生物聚合物，因为它们与megamolecular权重分级大分子的典型例子，它们的分子间相互作用使他们能够形成LC状态。

<table><tbody><tr><td>sacran</td><td>Green Science Materials Inc., Japan</td><td></td><td>From Aphanothece sacrum.&lt;br/&gt; M&lt;sub&gt;w&lt;/sub&gt; = 1.9 &amp;times; 10&lt;sup&gt;7&lt;/sup&gt; g mol&lt;sup&gt;-1&lt;/sup&gt;</td></tr><tr><td>xanthan gum</td><td>Taiyo Kagaku Co., Japan</td><td>Neosoft XC</td><td>From Xanthomonas campestris.&lt;br/&gt; M&lt;sub&gt;w&lt;/sub&gt; = 4.7 &amp;times; 10&lt;sup&gt;6&lt;/sup&gt; g mol&lt;sup&gt;-1&lt;/sup&gt;</td></tr><tr><td>tubulin</td><td>Cytoskeleton, Inc., USA</td><td>T240</td><td>From porcine brain.</td></tr><tr><td>GpCpp</td><td>Jena Bioscience, Germany</td><td>NU405L</td><td></td></tr><tr><td>piperazine-&lt;em&gt;N,N&amp;prime;&lt;/em&gt;-bis(2-ethanesulfonic acid)</td><td>Sigma-Aldrichi, Co. LLC.</td><td>P6757-500G</td><td>PIPES</td></tr><tr><td>ethylene glycol-bis(&amp;beta;-aminoethyl ether)-&lt;em&gt;N,N,N&#039;,N&#039;&lt;/em&gt;-tetraacetic acid</td><td>Dojindo Molecular Technologies, Inc.</td><td>342-01314</td><td>EGTA</td></tr><tr><td>MgCl&lt;sub&gt;2&lt;/sub&gt;-6H&lt;sub&gt;2&lt;/sub&gt;O</td><td>Wako Pure Chemical Industries, Ltd.</td><td>135-15055</td><td></td></tr><tr><td>KOH</td><td>Wako Pure Chemical Industries, Ltd.</td><td>162-21813</td><td>pellet</td></tr><tr><td>DNA</td><td>Sigma-Aldrich, Co. LLC.</td><td>D1626</td><td>From salmon testes.&lt;br/&gt;Mw = 1.3 &amp;times; 10&lt;sup&gt;6&lt;/sup&gt; Da (~2,000 bp)</td></tr><tr><td>Tris-EDTA buffer solution</td><td>Sigma-Aldrich, Co. LLC.</td><td>T9285-100ML</td><td>10 mM Tris, pH 8.0, with 1 mM EDTA</td></tr><tr><td>slide glass</td><td>Matsunami Glass Ind., Ltd., Japan</td><td>S1111</td><td></td></tr><tr><td>silicon rubber sheet</td><td>Asone Co.</td><td>6-611-32</td><td>Thickness: 1 mm</td></tr><tr><td>centrifuge</td><td>Beckman-Coulter, Inc., USA</td><td>Avanti J-25</td><td>equipped with a JA-20 rotor</td></tr><tr><td>light source</td><td>Sumita Optical Glass, Inc., Japan</td><td>LS-LHA</td><td></td></tr><tr><td>light guide</td><td>Sumita Optical Glass, Inc., Japan</td><td>GF7.2-1-L1500R-M80 (AAAR-015M)</td><td>80 mm &amp;times; 80 mm</td></tr><tr><td>halogen lamp</td><td>Ushio Inc., Japan</td><td>JCR 15V150WBN</td><td></td></tr><tr><td>holder</td><td>Sigmakoki, Co.,Ltd.</td><td>KMH-80</td><td></td></tr><tr><td>sample stage</td><td>Sigmakoki, Co.,Ltd.</td><td>TARW-25503L</td><td></td></tr><tr><td>sample holder</td><td>Sigmakoki, Co.,Ltd.</td><td>SHA-25RO</td><td></td></tr><tr><td>rod</td><td>Sigmakoki, Co.,Ltd.</td><td>ROU-12-40</td><td></td></tr><tr><td>posts holder</td><td>Sigmakoki, Co.,Ltd.</td><td>RS-6-40</td><td></td></tr><tr><td>posts holder</td><td>Sigmakoki, Co.,Ltd.</td><td>RS-12-60</td><td></td></tr><tr><td>posts holder</td><td>Sigmakoki, Co.,Ltd.</td><td>RS-12-80</td><td></td></tr><tr><td>posts holder</td><td>Sigmakoki, Co.,Ltd.</td><td>RS-12-130</td><td></td></tr><tr><td>carrier</td><td>Sigmakoki, Co.,Ltd.</td><td>CAA-25LS</td><td></td></tr><tr><td>camera holder</td><td>Sigmakoki, Co.,Ltd.</td><td>CMH-2</td><td></td></tr><tr><td>medium optical rail</td><td>Sigmakoki, Co.,Ltd.</td><td>OBA-500SH</td><td></td></tr><tr><td>lenstube</td><td>Tomytech, BORG</td><td>lenstube BK</td><td>80&amp;phi;, L25 mm&amp;nbsp;</td></tr><tr><td>lenstube</td><td>Tomytech, BORG</td><td>lenstube BK</td><td>80&amp;phi;, L50 mm&amp;nbsp;</td></tr><tr><td>multiband</td><td>Tomytech, BORG</td><td></td><td>80&amp;phi;&amp;nbsp;</td></tr><tr><td>V plate</td><td>Tomytech, BORG</td><td>V plate 60S</td><td></td></tr><tr><td>plate holder</td><td>Viexen, Co.,Ltd.</td><td>plate holder SX</td><td></td></tr><tr><td>EOS Kiss X7i&amp;nbsp;</td><td>Canon Inc., Japan</td><td>8594B001</td><td>with a standard zoom lens,&amp;nbsp; EFP 18-55 mm</td></tr><tr><td>photographic software</td><td>Canon Inc., Japan</td><td>EOS Utility</td><td></td></tr><tr><td>PC</td><td>Microsoft</td><td>Surface</td><td></td></tr><tr><td>polarization microscope</td><td>Olympus</td><td>BX51</td><td></td></tr><tr><td>first order retardation plate</td><td>Olympus</td><td>U-TP530</td><td>&amp;lambda; = 530 nm</td></tr><tr><td>CCD camera</td><td>Olympus</td><td>DP80</td><td></td></tr><tr><td>photographic software</td><td>Olympus</td><td>cellSens Standard</td><td></td></tr><tr><td>Java-based image processing program</td><td>the National Institutes of Health</td><td>ImageJ</td><td></td></tr></tbody></table>

methods for the self-integration of megamolecular biopolymers on the drying air-lc interface

使用水生物体总是容易在环境干燥。他们的活动具有基于生物聚合物 - 微观和宏观结构驱动，如在维管束移动水和皮肤保湿层的水的情况下看到。在这项研究中，我们开发了用于评估在干燥的生物聚合物组成的水性液晶（LC）的解决方案的效果的方法。由于LC生物聚合物具有megamolecular重，我们选择了研究多糖，细胞骨架蛋白和DNA。生物聚合物溶液的下偏振光干燥时的观测揭示milliscale自我整合从不稳定的空气-LC界面开始。水性LC生物聚合物溶液的动力学可以通过从单侧开孔蒸发水进行监测。通过分析使用交叉偏振光拍摄的图像，可以识别在取向秩序参数时空变化。这个方法可以不仅人工材料的各个领域，也自然的活组织的特性是有用的。我们相信，它将为软质材料在生物医学和环境领域提供的评价方法。

<html>使用该装置， 如图1中 ，从微区的自我整合到宏畴上的干燥空气-LC接口进行了评价（ 图2A）。作为干燥实验的第一示范，二种megamolecular多糖，SACRAN（M W = 1.9×107 克摩尔-1）和黄原胶（4.7×106 克摩尔-1）的，进行了比较。 图2B示出了在小区中的解决方案下的交叉偏振光的照片。干燥之前，它是可能观察到与透射光几个明亮区域的分散状态这两种解决方案。在60℃下干燥6个小时后，在溶液SACRAN空气-LC界面下方的区域成为高显著。这意味着，在结构域的凝胶收缩过程形成在界面宏观定向结构，类似于皮肤层形成"外部参照"&gt; 10。在另一方面，黄原胶溶液的强度，当温度刚从25℃增加至60℃，观察到的界面之下的一些小macrodomains急剧下降。这是因为黄原胶微区的流动性更为敏感温度比SACRAN微区的。界面上的取向的临界差异从而通过使用偏振装置检测。 图2C和Movie S1示出了用于在SACRAN溶液的干燥过程中空间-时间分析的结果。透射光强度指示中，微区的取向顺序参数。周围的空气 - 液体界面的峰强度显著增加，并且厚度生长至〜2毫米。这些结果清楚地表明，微区启动定向从空中-LC接口的ð长成平行于该界面的milliscale域。从下偏振光这种观察，因此可以以可视化的流体运动中的干燥过程。 在宏畴尺寸和取向方向而言SACRAN和黄原胶之间的差异也通过用一阶延迟板（ 图2D）的偏光显微镜证实。该SACRAN流体相显示一个蓝色区域，这意味着形成在界面处的单个宏畴。与此相反，黄原胶流体相呈蓝色，黄色和粉红色的区域，这意味着与任意方向形成多个macrodomains。 该方法还探讨用于与megamolecular权重，比较了三种类型的基本刚性的生物聚合物的-多糖（SACRAN:M W = 1.9×107 克摩尔-1，微区长度&gt; 20微米），的MT（M W = 10 9 -1010克摩尔-1，微区长度&gt; 10微米），和DNA（M W = 1.3×106 克摩尔-1，微区长度&lt;1微米） -在37℃的生理环境。如在图3B中所示的干燥之前，SACRAN溶液和MT溶液显示出相似的LC状态，其中在整个区域散射结构域。在干燥过程中，在MT溶液中的结构域也经历自我整合从空气-LC接口，和散射结构域集成到单个宏畴。在另一方面，该DNA溶液显示在液相中没有特定取向。在MT溶液，并用缓冲液中制备的DNA溶液的情况下，需要注意的是集成是通过在干燥过程中变化，盐浓度，pH和离子强度的影响是重要的。 图3C）。所述SACRAN干燥记录显示出因为milliscale单个宏畴形成的显著双折射强度。用于MT干燥记录，我们观察到波状束，其中长轴平行于X轴。与此相反，DNA溶液的干燥记录显示晶粒形状macrodomains &lt;5微米的直径和在任意方向。从这些观察，很清楚的是，宏畴尺寸由棒状微区的长度的影响。这是因此，可以通过使用偏振显微镜观察干燥记录来评估生物聚合物微区的方向取向。 总之，使用使用偏振光的方法对干燥空气-LC接口megamolecular生物聚合物的自我整合。水性LC溶液的动力学是由从一侧开细胞蒸发水监测。通过分析使用交叉偏振光拍摄的图像，它是能够识别时空变化和取向有序参数。考虑到表现出干燥过程类似于自然过程，这种方法将是不仅人工材料在各种领域中的表征是有用的，但也自然生活组织。我们认为，这可能为软质材料在生物医学和环境领域提供的评价方法。 <img alt="图1" src="/files/ftp_upload/55274/55274fig1.jpg" /> 图1:干燥实验。 （A）在一侧开孔的解决方案的示意图。 （B）下交叉偏振光用于观测实验装置的示意图。偏振片通常被调整为45°和135°。 （C） 实际的设备。该图已被从参照图6进行修改。版权所有:美国化学学会，2016年<a href="https://ecsource-jove-com.bdigitaluss.remotexs.co/files/ftp_upload/55274/55274fig1large.jpg" target="_blank">请点击此处查看该图的放大版本。</a> <img alt="图2" src="/files/ftp_upload/55274/55274fig2.jpg" /> 图2:多糖SACRAN和黄原胶的水溶液的干燥过程。 （A） 干燥空气-LC接口上的自我整合的示意图。 （B） 在初始状态后2种多糖溶液的侧视图在60℃下交偏振光干燥6个小时。初始聚合物浓度为0.5重量％。 （C） 通过正交尼科耳上用于在给定时间拍摄每个画面的Z方向的直线透射光强度。 （D） 在单元中的聚合物溶液在60℃下干燥6个小时后的偏光显微镜图像。红色箭头:在接口上macrodomains。该图已被从参照图6进行修改。版权所有:美国化学学会，2016年<a href="https://ecsource-jove-com.bdigitaluss.remotexs.co/files/ftp_upload/55274/55274fig2large.jpg" target="_blank">请点击此处查看该图的放大版本。</a> <img alt="图3" src="/files/ftp_upload/55274/55274fig3.jpg" /> 图3:生物聚合物的水溶液和干燥记录的干燥过程。 （A）的多边形的来源accharides（蓝藻）的MT（猪脑）和DNA（鲑鱼睾丸）。 （B） 在37℃下交偏振光干燥时SACRAN，MT，和DNA溶液的侧视图。初始聚合物浓度为0.5重量％。 （C） 通过正交尼科耳的给定方向上的玻璃基板上的干燥的聚合物膜的显微图像。所有比例尺是50微米。该图已被从参照图6进行修改。版权所有:美国化学学会，2016年<a href="https://ecsource-jove-com.bdigitaluss.remotexs.co/files/ftp_upload/55274/55274fig3large.jpg" target="_blank">请点击此处查看该图的放大版本。</a> <img alt="电影S1" src="/files/ftp_upload/55274/55274vid1.jpg" /> 电影S1:在60℃ 下 干燥时干燥所述SACRAN溶液的过程中 ，使用不良观察交叉尼科尔。 <a href="https://ecsource-jove-com.bdigitaluss.remotexs.co/files/ftp_upload/55274/Movie_S1-3fps.avi" target="_blank">请点击这里下载这部电影。</a>

Watch this Scientific Journal Video about Methods for the Self-integration of Megamolecular Biopolymers on the Drying Air-LC Interface at JoVE.com

Methods for the Self-integration of Megamolecular Biopolymers on the Drying Air-LC Interface

在这里提供了一种用于生物聚合物megamolecular的空气 - 液体界面的结晶在干燥引起的自我整合方法。这种方法将是有益的，不仅对于理解生物聚合物的宏观潜力，同时也为软质材料在生物医学和环境领域的评价方法。

对干燥空气-LC接口Megamolecular生物大分子的自我整合方法

Living organisms that use water are always prone to drying in the environment. Their activities are driven by biopolymer-based micro- and ...

methods-for-self-integration-megamolecular-biopolymers-on-drying-air

Universidad San Sebastian

Research

JoVE Journal

Bioengineering

6.0K 次观看.  Japan Advanced Institute of Science and Technology. 在这里提供了一种用于生物聚合物megamolecular的空气 - 液体界面的结晶在干燥引起的自我整合方法。这种方法将是有益的，不仅对于理解生物聚合物的宏观潜力，同时也为软质材料在生物医学和环境领域的评价方法。 

视频: Methods for the Self-integration of Megamolecular Biopolymers on the Drying Air-LC Interface

Synthesis of Poly(N-isopropylacrylamide) Janus Microhydrogels for Anisotropic Thermo-responsiveness and Organophilic/Hydrophilic Loading Capability

Janus microparticles are compartmentalized particles with differing molecular structures and/or functionality on each of their two sides. Because of this unique property, Janus microparticles have been recognized as a new class of materials, thereby attracting a great deal of attention from various research fields. The versatility of these microparticles has been exemplified through their uses as building blocks for self-assembly, electrically responsive actuators, emulsifiers for painting and cosmetics, and carriers for drug delivery. This study introduces a detailed protocol that explicitly describes a synthetic method for designing novel Janus microhydrogels composed of a single base material, poly(N-isopropylacrylamide) (PNIPAAm). Janus microdroplets are firstly generated via a hydrodynamic focusing microfluidic device (HFMD) based on the separation of a supersaturated aqueous NIPAAm monomer solution and subsequently polymerized through exposure to UV irradiation. The resulting Janus microhydrogels were found to be entirely composed of the same base material, featured an easily identifiable compartmentalized morphology, and exhibited anisotropic thermo-responsiveness and organophilic/hydrophilic loading capability. We believe that the proposed method introduces a novel hydrogel platform with the potential for advanced synthesis of multi-functional Janus microhydrogels.

Synthesis of PolyN-isopropylacrylamide Janus Microhydrogels for Anisotropic Thermo-responsiveness and Organophilic/Hydrophilic Loading Capability

We present a protocol to synthesize Janus microhydrogels composed entirely of the same base material, poly(N-isopropylacrylamide) (PNIPAAm), with a clearly compartmentalized structure base on the phase separation of a supersaturated NIPAAm monomer solution. The synthesized Janus microhydrogels show unique properties such as anisotropic thermo-responsiveness and organophilic/hydrophilic loading capability.

合成聚（<em&gt;ñ</em&gt; -isopropylacrylamide）剑锋Microhydrogels各向异性热响应能力和亲有机/亲水承载能力

Janus microparticles are compartmentalized particles with differing molecular structures and/or functionality on each of their two sides. Because of this ...

科研

化学

Orientational Transition in a Liquid Crystal Triggered by the Thermodynamic Growth of Interfacial Wetting Sheets

In liquid crystal (LC) physical chemistry, molecules near the surface play a great role in controlling bulk orientation. Thus far, mainly to achieve desired molecular orientation states in LC displays, the "static" surface property of LCs, so-called surface anchoring, has been intensively studied. As a rule of thumb, once the initial orientation of LCs is "locked" by specific surface treatments, such as rubbing or treatment with a specific alignment layer, it hardly changes with temperature. Here, we present a system exhibiting an orientational transition upon temperature variation, which conflicts with the consensus. Right on the transition, the bulk LC molecules experience the orientational rotation, with 90&#176; between the planar (P) orientation at high temperatures and the vertical (V) orientation at low temperatures in the first-order transitional manner. We have tracked thermodynamic surface anchoring behavior by means of polarizing optical microscopy (POM), dielectric spectroscopy (DS), high-resolution differential scanning calorimetry (HR-DSC), and grazing incidence X-ray diffraction (GI-XRD) and reached a plausible physical explanation: that the transition is triggered by a growth of surface wetting sheets, which impose the V orientation locally against the P orientation in the bulk. This landscape would provide a general link explaining how the equilibrium bulk orientation is affected by surface-localized orientation in many LC systems. In our characterization, POM and DS are advantageous by offering information on the spatial distribution of the orientation of LC molecules. HR-DSC gives information about the precise thermodynamic information on transitions, which cannot be addressed by conventional DSC instruments due to limited resolution. GI-XRD provides information on surface-specific molecular orientation and short-range orderings. The goal of this paper is to present a protocol for preparing a sample that exhibits the transition and to demonstrate how the thermodynamic structural variation, both in the bulk and on surfaces, can be analyzed through the abovementioned methods.

Here, we present a protocol to trigger an orientational transition of a liquid crystal in response to temperature. Methodologies are described for preparing a sample in order to observe the transition and the detailed transitional evolution.

由界面润湿片的热力学生长触发的液晶中的定向转变

In liquid crystal (LC) physical chemistry, molecules near the surface play a great role in controlling bulk orientation. Thus far, mainly to achieve ...

工程学

Interfacial Molecular-level Structures of Polymers and Biomacromolecules Revealed via Sum Frequency Generation Vibrational Spectroscopy

As a second-order nonlinear optical spectroscopy, sum frequency generation (SFG) vibrational spectroscopy has widely been used in investigating various surfaces and interfaces. This non-invasive optical technique can provide the local molecular-level information with monolayer or submonolayer sensitivity. We here are providing experimental methodology on how to selectively detect the buried interface for both macromolecules and biomacromolecules. With this in mind, interfacial secondary structures of silk fibroin and water structures around model short-chain oligonucleotide duplex are discussed. The former shows a chain-chain overlap or spatial confinement effect and the latter shows a protection function against the Ca2+ ions resulting from the chiral spine superstructure of water.

Being comprehensively utilized, sum frequency generation (SFG) vibrational spectroscopy can help to reveal chain conformational order and secondary structural change happening at polymer and biomacromolecule interfaces.

通过相频生成振动光谱揭示聚合物和生物大分子的界面分子级结构

As a second-order nonlinear optical spectroscopy, sum frequency generation (SFG) vibrational spectroscopy has widely been used in investigating various ...

生物化学

Preparation of Liquid Crystal Networks for Macroscopic Oscillatory Motion Induced by Light

A strategy based on doped liquid crystalline networks is described to create mechanical self-sustained oscillations of plastic films under continuous light irradiation. The photo-excitation of dopants that can quickly dissipate light into heat, coupled with anisotropic thermal expansion and self-shadowing of the film, gives rise to the self-sustained deformation. The oscillations observed are influenced by the dimensions and the modulus of the film, and by the directionality and intensity of the light. The system developed offers applications in energy conversion and harvesting for soft-robotics and automated systems.
The general method described here consists of creating free-standing liquid crystalline films and characterizing the mechanical and thermal effects observed. The molecular alignment is achieved using alignment layers (rubbed polyimide), commonly used in the display manufacturing industry. To obtain actuators with large deformation, the mesogens are aligned and polymerized in a splay/bend configuration, i.e., with the director of the liquid crystals (LCs) going gradually from planar to homeotropic through the film thickness. Upon irradiation, the mechanical and thermal oscillations obtained are monitored with a high-speed camera. The results are further quantified by image analysis using an image processing program.

The goal of the protocol is to create liquid crystalline polymer films that can mechanically oscillate under continuous light irradiation. We describe in great detail the conception of free-standing films, from the liquid crystal alignment method to the photo-actuation. The experimental protocol applied to prepare this material is broadly applicable.

光诱导的宏观振荡运动液晶网络的制备

A strategy based on doped liquid crystalline networks is described to create mechanical self-sustained oscillations of plastic films under continuous ...

Microfluidic Preparation of Liquid Crystalline Elastomer Actuators

This paper focuses on the microfluidic process (and its parameters) to prepare actuating particles from liquid crystalline elastomers. The preparation usually consists in the formation of droplets containing low molar mass liquid crystals at elevated temperatures. Subsequently, these particle precursors are oriented in the flow field of the capillary and solidified by a crosslinking polymerization, which produces the final actuating particles. The optimization of the process is necessary to obtain the actuating particles and the proper variation of the process parameters (temperature and flow rate) and allows variations of size and shape (from oblate to strongly prolate morphologies) as well as the magnitude of actuation. In addition, it is possible to vary the type of actuation from elongation to contraction depending on the director profile induced to the droplets during the flow in the capillary, which again depends on the microfluidic process and its parameters. Furthermore, particles of more complex shapes, like core-shell structures or Janus particles, can be prepared by adjusting the setup. By the variation of the chemical structure and the mode of crosslinking (solidification) of the liquid crystalline elastomer, it is also possible to prepare actuating particles triggered by heat or UV-vis irradiation.

This article describes the microfluidic process and parameters to prepare actuating particles from liquid crystalline elastomers. This process allows the preparation of actuating particles and the variation of their size and shape (from oblate to strongly prolate, core-shell, and Janus morphologies) as well as the magnitude of actuation.

液晶弹性体致动器的微流控制备

This paper focuses on the microfluidic process (and its parameters) to prepare actuating particles from liquid crystalline elastomers. The preparation ...

Preparation of Monodomain Liquid Crystal Elastomers and Liquid Crystal Elastomer Nanocomposites

LCEs are shape-responsive materials with fully reversible shape change and potential applications in medicine, tissue engineering, artificial muscles, and as soft robots. Here, we demonstrate the preparation of shape-responsive liquid crystal elastomers (LCEs) and LCE nanocomposites along with characterization of their shape-responsiveness, mechanical properties, and microstructure. Two types of LCEs &#8212; polysiloxane-based and epoxy-based &#8212; are synthesized, aligned, and characterized. Polysiloxane-based LCEs are prepared through two crosslinking steps, the second under an applied load, resulting in monodomain LCEs. Polysiloxane LCE nanocomposites are prepared through the addition of conductive carbon black nanoparticles, both throughout the bulk of the LCE and to the LCE surface. Epoxy-based LCEs are prepared through a reversible esterification reaction. Epoxy-based LCEs are aligned through the application of a uniaxial load at elevated (160 &#176;C) temperatures. Aligned LCEs and LCE nanocomposites are characterized with respect to reversible strain, mechanical stiffness, and liquid crystal ordering using a combination of imaging, two-dimensional X-ray diffraction measurements, differential scanning calorimetry, and dynamic mechanical analysis. LCEs and LCE nanocomposites can be stimulated with heat and/or electrical potential to controllably generate strains in cell culture media, and we demonstrate the application of LCEs as shape-responsive substrates for cell culture using a custom-made apparatus.

We demonstrate the preparation of siloxane-based and epoxy-based liquid crystal elastomers (LCEs) and LCE nanocomposites. The LCEs are characterized with respect to reversible strain, liquid crystal ordering, and stiffness. As a potential application, we demonstrate their use as shape-responsive substrates in a custom device for active cell culture.

单域液晶的弹性体和液晶橡胶纳米复合材料的制备

LCEs are shape-responsive materials with fully reversible shape change and potential applications in medicine, tissue engineering, artificial muscles, and ...

生物工程

Automated Lipid Bilayer Membrane Formation Using a Polydimethylsiloxane Thin Film

An artificial lipid bilayer, or black lipid membrane (BLM), is a powerful tool for studying ion channels and protein interactions, as well as for biosensor applications. However, conventional BLM formation techniques have several drawbacks and they often require specific expertise and laborious processes. In particular, conventional BLMs suffer from low formation success rates and inconsistent membrane formation time. Here, we demonstrate a storable and transportable BLM formation system with controlled thinning-out time and enhanced BLM formation rate by replacing conventionally used films (polytetrafluoroethylene, polyoxymethylene, polystyrene) to polydimethylsiloxane (PDMS). In this experiment, a porous-structured polymer such as PDMS thin film is used. In addition, as opposed to conventionally used solvents with low viscosity, the use of squalene permitted a controlled thinning-out time via slow solvent absorption by PDMS, prolonging membrane lifetime. In addition, by using a mixture of squalene and hexadecane, the freezing point of the lipid solution was increased (~16 &#176;C), in addition, membrane precursors were produced that can be indefinitely stored and readily transported. These membrane precursors have reduced BLM formation time of &#60; 1 hr and achieved a BLM formation rate of ~80%. Moreover, ion channel experiments with gramicidin A demonstrated the feasibility of the membrane system.

We demonstrate a storable, transportable lipid bilayer formation system. A lipid bilayer membrane can be formed within 1 hr with over 80% success rate when a frozen membrane precursor is brought to ambient temperature. This system will reduce laborious processes and expertise associated with ion channels.

自动化脂双层膜的形成使用聚二甲基硅氧烷薄膜

An artificial lipid bilayer, or black lipid membrane (BLM), is a powerful tool for studying ion channels and protein interactions, as well as for ...

Self-Assembly of Hybrid Lipid Membranes Doped with Hydrophobic Organic Molecules at the Water/Air Interface

Because of their unique properties, including an ultrathin thickness (3-4 nm), ultrahigh resistivity, fluidity and self-assembly ability, lipid bilayers can be readily functionalized and have been used in various applications such as bio-sensors and bio-devices. In this study, we introduced a planar organic molecule: copper (II) 2,9,16,23-tetra-tert-butyl-29H,31H-phthalocyanine (CuPc) to dope lipid membranes. The CuPc/lipid hybrid membrane forms at the water/air interface by self-assembly. In this membrane, the hydrophobic CuPc molecules are located between the hydrophobic tails of lipid molecules, forming a lipid/CuPc/lipid sandwich structure. Interestingly, an air-stable hybrid lipid bilayer can be readily formed by transferring the hybrid membrane onto a Si substrate. We report a straightforward method for incorporating nanomaterials into a lipid bilayer system, which represents a new methodology for the fabrication of biosensors and biodevices.

We report a protocol for producing a hybrid lipid membrane at the water/air interface by doping the lipid bilayer with copper (II) 2,9,16,23-tetra-tert-butyl-29H,31H-phthalocyanine (CuPc) molecules. The resulting hybrid lipid membrane has a lipid/CuPc/lipid sandwich structure. This protocol can also be applied to the formation of other functional nanomaterials.

在水/空气界面上掺杂与疏水有机分子混合脂膜的自组装

Because of their unique properties, including an ultrathin thickness (3-4 nm), ultrahigh resistivity, fluidity and self-assembly ability, lipid bilayers ...

Crystallization of Proteins on Chip by Microdialysis for In Situ X-ray Diffraction Studies

This protocol describes the manufacturing of reproducible and inexpensive microfluidic devices covering the whole pipeline for crystallizing proteins on-chip with the dialysis method and allowing in situ single-crystal or serial crystallography experiments at room temperature. The protocol details the fabrication process of the microchips, the manipulation of the on-chip crystallization experiments and the treatment of the in situ collected X-ray diffraction data for the structural elucidation of the protein sample. The main feature of this microfabrication procedure lies on the integration of a commercially available, semipermeable regenerated cellulose dialysis membrane in between two layers of the chip. The molecular weight cut-off of the embedded membrane varies depending on the molecular weight of the macromolecule and the precipitants. The device exploits the advantages of microfluidic technology, such as the use of minute volumes of samples (&#60;1 &#181;L) and fine tuning over transport phenomena. The chip coupled them with the dialysis method, providing precise and reversible control over the crystallization process and can be used for investigating phase diagrams of proteins at the microliter scale. The device is patterned using a photocurable thiolene-based resin with soft imprint lithography on an optically transparent polymeric substrate. Moreover, the background scattering of the materials composing the microchips and generating background noise was evaluated rendering the chip compatible for in situ X-ray diffraction experiments. Once protein crystals are grown on-chip up to an adequate size and population uniformity, the microchips can be directly mounted in front of the X-ray beam with the aid of a 3D printed holder. This approach addresses the challenges rising from the use of cryoprotectants and manual harvesting in conventional protein crystallography experiments through an easy and inexpensive manner. Complete X-ray diffraction data sets from multiple, isomorphous lysozyme crystals grown on-chip were collected at room temperature for structure determination.

Crystallization of Proteins on Chip by Microdialysis for In Situ X-ray Diffraction Studies

This paper details the fabrication protocol of microfluidic chips developed for on-chip protein crystallization with the dialysis method and in situ X-ray diffraction experiments. The microfabrication process makes it possible to integrate a semipermeable regenerated cellulose dialysis membrane with any molecular weight cut-off, between two layers of the chip.

通过微透析对芯片上的蛋白质进行结晶，用于 原位 X射线衍射研究

This protocol describes the manufacturing of reproducible and inexpensive microfluidic devices covering the whole pipeline for crystallizing proteins ...

Crystallization of Membrane Proteins in Lipidic Mesophases

Membrane proteins perform critical functions in living cells related to signal transduction, transport and energy transformations, and, as such, are implicated in a multitude of malfunctions and diseases. However, a structural and functional understanding of membrane proteins is strongly lagging behind that of their soluble partners, mainly, due to difficulties associated with their solubilization and generation of diffraction quality crystals. Crystallization in lipidic mesophases (also known as in meso or LCP crystallization) is a promising technique which was successfully applied to obtain high resolution structures of microbial rhodopsins, photosynthetic proteins, outer membrane beta barrels and G protein-coupled receptors. In meso crystallization takes advantage of a native-like membrane environment and typically produces crystals with lower solvent content and better ordering as compared to traditional crystallization from detergent solutions. The method is not difficult, but requires an understanding of lipid phase behavior and practice in handling viscous mesophase materials. Here we demonstrate a simple and efficient way of making LCP and reconstituting a membrane protein in the lipid bilayer of LCP using a syringe mixer, followed by dispensing nanoliter portions of LCP into an assay or crystallization plate, conducting pre-crystallization assays and harvesting crystals from the LCP matrix. These protocols provide a basic guide for approaching in meso crystallization trials; however, as with any crystallization experiment, extensive screening and optimization are required, and a successful outcome is not necessarily guaranteed.

The protocols describe the essential steps for obtaining diffraction quality crystals of a membrane protein starting from reconstitution of the protein in a lipidic cubic phase (LCP), finding initial conditions with LCP-FRAP pre-crystallization assays, setting up LCP crystallization trials and harvesting crystals.

在油脂介孔膜蛋白的结晶

Membrane proteins perform critical functions in living cells related to signal transduction, transport and energy transformations, and, as such, are ...

对干燥空气-LC接口Megamolecular生物大分子的自我整合方法

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合成聚（

由界面润湿片的热力学生长触发的液晶中的定向转变

通过相频生成振动光谱揭示聚合物和生物大分子的界面分子级结构

光诱导的宏观振荡运动液晶网络的制备

液晶弹性体致动器的微流控制备

单域液晶的弹性体和液晶橡胶纳米复合材料的制备

自动化脂双层膜的形成使用聚二甲基硅氧烷薄膜

在水/空气界面上掺杂与疏水有机分子混合脂膜的自组装

通过微透析对芯片上的蛋白质进行结晶，用于原位 X射线衍射研究

在油脂介孔膜蛋白的结晶

对干燥空气-LC接口Megamolecular生物大分子的自我整合方法

摘要

探索更多视频

此视频中的章节

合成聚（

由界面润湿片的热力学生长触发的液晶中的定向转变

通过相频生成振动光谱揭示聚合物和生物大分子的界面分子级结构

光诱导的宏观振荡运动液晶网络的制备

液晶弹性体致动器的微流控制备

单域液晶的弹性体和液晶橡胶纳米复合材料的制备

自动化脂双层膜的形成使用聚二甲基硅氧烷薄膜

在水/空气界面上掺杂与疏水有机分子混合脂膜的自组装

通过微透析对芯片上的蛋白质进行结晶，用于 原位 X射线衍射研究

在油脂介孔膜蛋白的结晶

通过微透析对芯片上的蛋白质进行结晶，用于原位 X射线衍射研究