Name: methods for the self-integration of megamolecular biopolymers on the drying air-lc interface
Uploaded: 2017-04-07T12:30:00
Description: Watch this Scientific Journal Video about Methods for the Self-integration of Megamolecular Biopolymers on the Drying Air-LC Interface at JoVE.com

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）?...

<ol>
	<li>Flemming, H. -. C., Wingender, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+biofilm+matrix.">The biofilm matrix.</a> Nat. Rev. Microbiology. 8 (9), 623-633 (2010).</li><li>Osada, Y., Kawamura, R., Sano, K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> Hydrogels of cytoskeletal proteins. , (2016).</li><li>Rothemund, P. W. K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Folding+DNA+to+create+nanoscale+shapes+and+patterns.">Folding DNA to create nanoscale shapes and patterns.</a> Nature. 440, 297-302 (2006).</li><li>Okeyoshi, K., Okajima, M. K., Kaneko, T. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Milliscale+self-integration+of+megamolecule+biopolymers+on+a+drying+gas-aqueous+liquid+crystalline+interface.">Milliscale self-integration of megamolecule biopolymers on a drying gas-aqueous liquid crystalline interface.</a> Biomacromolecules. 17 (6), 2096-2103 (2016).</li><li>De Gennes, P. G., Prost, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> The physics of liquid crystals. , (1993).</li><li>De Gennes, P. G., Brochard-Wyart, F., Quere, D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> Capillarity and wetting phenomena: drops, bubbles, pearls, waves. , (2003).</li><li>Okajima, M. K., Kaneko, D., Mitsumata, T., Kaneko, T., Watanabe, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Cyanobacteria+that+produce+megamolecules+with+efficient+self-orientations.">Cyanobacteria that produce megamolecules with efficient self-orientations.</a> Macromolecules. 42 (8), 3058-3062 (2009).</li><li>Okeyoshi, K., Kawamura, R., Yoshida, R., Osada, Y. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Thermo-+and+photo-enhanced+microtubule+formation+from+Ru(bpy)32+-conjugated+tubulin.">Thermo- and photo-enhanced microtubule formation from Ru(bpy)32+-conjugated tubulin.</a> J. Mater. Chem. B. 2, 41-45 (2014).</li><li>Matsuo, E. S., Tanaka, T. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Kinetics+of+discontinuous+volume-phase+transition+of+gels.">Kinetics of discontinuous volume-phase transition of gels.</a> J. Chem. Phys. 89, 1695-1703 (1988).</li><li>Okajima, K., Mishima, R., Amornwachirabodee, K., Mitsumata, T., Okeyoshi, K., Kaneko, T. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Anisotropic+swelling+in+hydrogels+formed+by+cooperatively+aligned+megamolecules.">Anisotropic swelling in hydrogels formed by cooperatively aligned megamolecules.</a> RSC Adv. 5, 86723-86729 (2015).</li><li>Joshi, G., Okeyoshi, K., Okajima, M. K., Kaneko, T. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Directional+control+of+diffusion+and+swelling+in+megamolecular+polysaccharide+hydrogels.">Directional control of diffusion and swelling in megamolecular polysaccharide hydrogels.</a> Soft Matter. 12, 5515-5518 (2016).</li></ol>

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。?...

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 border="0" cellpadding="0" cellspacing="0" width="617">
	<tbody>
		<tr height="69">
			<td height="69" style="height:69px;width:172px;">sacran</td>
			<td style="width:151px;">Green Science Materials Inc., Japan</td>
			<td style="width:131px;"></td>
			<td style="width:163px;">From Aphanothece sacrum. 
			Mw = 1.9 &#215; 107 g mol-1</td>
		</tr>
		<tr height="69">
			<td height="69" style="height:69px;width:172px;">xanthan gum</td>
			<td style="width:151px;">Taiyo Kagaku Co., Japan</td>
			<td style="width:131px;">Neosoft XC</td>
			<td style="width:163px;">From Xanthomonas campestris. 
			Mw = 4.7 &#215; 106 g mol-1</td>
		</tr>
		<tr height="43">
			<td height="43" style="height:43px;width:172px;">tubulin</td>
			<td style="width:151px;">Cytoskeleton, Inc., USA</td>
			<td style="width:131px;">T240</td>
			<td style="width:163px;">From porcine brain.</td>
		</tr>
		<tr height="43">
			<td height="43" style="height:43px;width:172px;">GpCpp</td>
			<td style="width:151px;">Jena Bioscience, Germany</td>
			<td style="width:131px;">NU405L</td>
			<td style="width:163px;"></td>
		</tr>
		<tr height="43">
			<td height="43" style="height:43px;width:172px;">piperazine-N,N&#8242;-bis(2-ethanesulfonic acid)</td>
			<td style="width:151px;">Sigma-Aldrichi, Co. LLC.</td>
			<td style="width:131px;">P6757-500G</td>
			<td style="width:163px;">PIPES</td>
		</tr>
		<tr height="85">
			<td height="85" style="height:85px;width:172px;">ethylene glycol-bis(&#946;-aminoethyl ether)-N,N,N&#39;,N&#39;-tetraacetic acid</td>
			<td style="width:151px;">Dojindo Molecular Technologies, Inc.</td>
			<td style="width:131px;">342-01314</td>
			<td style="width:163px;">EGTA</td>
		</tr>
		<tr height="43">
			<td height="43" style="height:43px;width:172px;">MgCl2-6H2O</td>
			<td style="width:151px;">Wako Pure Chemical Industries, Ltd.</td>
			<td style="width:131px;">135-15055</td>
			<td style="width:163px;"></td>
		</tr>
		<tr height="43">
			<td height="43" style="height:43px;width:172px;">KOH</td>
			<td style="width:151px;">Wako Pure Chemical Industries, Ltd.</td>
			<td style="width:131px;">162-21813</td>
			<td style="width:163px;">pellet</td>
		</tr>
		<tr height="21">
			<td height="58" style="height:58px;width:172px;">DNA</td>
			<td rowspan="2" style="width:151px;">Sigma-Aldrich, Co. LLC.</td>
			<td rowspan="2" style="width:131px;">D1626</td>
			<td rowspan="2" style="width:163px;">From salmon testes. 
			Mw = 1.3 &#215; 106 Da (~2000 bp)</td>
		</tr>
		<tr height="43">
			<td height="43" style="height:43px;width:172px;">Tris-EDTA buffer solution</td>
			<td style="width:151px;">Sigma-Aldrich, Co. LLC.</td>
			<td style="width:131px;">T9285-100ML</td>
			<td style="width:163px;">10 mM Tris, pH 8.0, with 1 mM EDTA</td>
		</tr>
		<tr height="43">
			<td height="43" style="height:43px;width:172px;">slide glass</td>
			<td style="width:151px;">Matsunami Glass Ind., Ltd., Japan</td>
			<td style="width:131px;">S1111</td>
			<td style="width:163px;"></td>
		</tr>
		<tr height="22">
			<td height="22" style="height:22px;width:172px;">silicon rubber sheet</td>
			<td style="width:151px;">Asone Co.</td>
			<td style="width:131px;">6-611-32</td>
			<td style="width:163px;">Thickness: 1 mm</td>
		</tr>
		<tr height="43">
			<td height="43" style="height:43px;width:172px;">centrifuge</td>
			<td style="width:151px;">Beckman-Coulter, Inc., USA</td>
			<td style="width:131px;">Avanti J-25</td>
			<td style="width:163px;">equipped with a JA-20 rotor</td>
		</tr>
		<tr height="43">
			<td height="43" style="height:43px;width:172px;">light source</td>
			<td style="width:151px;">Sumita Optical Glass, Inc., Japan</td>
			<td style="width:131px;">LS-LHA</td>
			<td style="width:163px;"></td>
		</tr>
		<tr height="43">
			<td height="43" style="height:43px;width:172px;">light guide</td>
			<td style="width:151px;">Sumita Optical Glass, Inc., Japan</td>
			<td style="width:131px;">GF7.2-1-L1500R-M80 (AAAR-015M)</td>
			<td style="width:163px;">80 mm &#215; 80 mm</td>
		</tr>
		<tr height="22">
			<td height="22" style="height:22px;width:172px;">halogen lamp</td>
			<td style="width:151px;">Ushio Inc., Japan</td>
			<td style="width:131px;">JCR 15V150WBN</td>
			<td style="width:163px;"></td>
		</tr>
		<tr height="22">
			<td height="22" style="height:22px;width:172px;">polarizer</td>
			<td style="width:151px;">Luceo, Co.,Ltd.</td>
			<td style="width:131px;">POLAX-42S</td>
			<td style="width:163px;">40 x 80 x 2t (45&#730;)</td>
		</tr>
		<tr height="22">
			<td height="22" style="height:22px;width:172px;">holder</td>
			<td style="width:151px;">Sigmakoki, Co.,Ltd.</td>
			<td style="width:131px;">KMH-80</td>
			<td style="width:163px;"></td>
		</tr>
		<tr height="22">
			<td height="22" style="height:22px;width:172px;">sample stage</td>
			<td style="width:151px;">Sigmakoki, Co.,Ltd.</td>
			<td style="width:131px;">TARW-25503L</td>
			<td style="width:163px;"></td>
		</tr>
		<tr height="22">
			<td height="22" style="height:22px;width:172px;">sample holder</td>
			<td style="width:151px;">Sigmakoki, Co.,Ltd.</td>
			<td style="width:131px;">SHA-25RO</td>
			<td style="width:163px;"></td>
		</tr>
		<tr height="22">
			<td height="22" style="height:22px;width:172px;">rod</td>
			<td style="width:151px;">Sigmakoki, Co.,Ltd.</td>
			<td style="width:131px;">ROU-12-40</td>
			<td style="width:163px;"></td>
		</tr>
		<tr height="22">
			<td height="22" style="height:22px;width:172px;">posts holder</td>
			<td style="width:151px;">Sigmakoki, Co.,Ltd.</td>
			<td style="width:131px;">RS-6-40</td>
			<td style="width:163px;"></td>
		</tr>
		<tr height="22">
			<td height="22" style="height:22px;width:172px;">posts holder</td>
			<td style="width:151px;">Sigmakoki, Co.,Ltd.</td>
			<td style="width:131px;">RS-12-60</td>
			<td style="width:163px;"></td>
		</tr>
		<tr height="22">
			<td height="22" style="height:22px;width:172px;">posts holder</td>
			<td style="width:151px;">Sigmakoki, Co.,Ltd.</td>
			<td style="width:131px;">RS-12-80</td>
			<td style="width:163px;"></td>
		</tr>
		<tr height="22">
			<td height="22" style="height:22px;width:172px;">posts holder</td>
			<td style="width:151px;">Sigmakoki, Co.,Ltd.</td>
			<td style="width:131px;">RS-12-130</td>
			<td style="width:163px;"></td>
		</tr>
		<tr height="22">
			<td height="22" style="height:22px;width:172px;">carrier</td>
			<td style="width:151px;">Sigmakoki, Co.,Ltd.</td>
			<td style="width:131px;">CAA-25LS</td>
			<td style="width:163px;"></td>
		</tr>
		<tr height="22">
			<td height="22" style="height:22px;width:172px;">camera holder</td>
			<td style="width:151px;">Sigmakoki, Co.,Ltd.</td>
			<td style="width:131px;">CMH-2</td>
			<td style="width:163px;"></td>
		</tr>
		<tr height="22">
			<td height="22" style="height:22px;width:172px;">medium optical rail</td>
			<td style="width:151px;">Sigmakoki, Co.,Ltd.</td>
			<td style="width:131px;">OBA-500SH</td>
			<td style="width:163px;"></td>
		</tr>
		<tr height="22">
			<td height="22" style="height:22px;width:172px;">lenstube</td>
			<td style="width:151px;">Tomytech, BORG</td>
			<td style="width:131px;">lenstube BK</td>
			<td style="width:163px;">80&#966;, L25 mm&#160;</td>
		</tr>
		<tr height="22">
			<td height="22" style="height:22px;width:172px;">lenstube</td>
			<td style="width:151px;">Tomytech, BORG</td>
			<td style="width:131px;">lenstube BK</td>
			<td style="width:163px;">80&#966;, L50 mm&#160;</td>
		</tr>
		<tr height="22">
			<td height="22" style="height:22px;width:172px;">multiband</td>
			<td style="width:151px;">Tomytech, BORG</td>
			<td style="width:131px;"></td>
			<td style="width:163px;">80&#966;&#160;</td>
		</tr>
		<tr height="22">
			<td height="22" style="height:22px;width:172px;">V plate</td>
			<td style="width:151px;">Tomytech, BORG</td>
			<td style="width:131px;">V plate 60S</td>
			<td style="width:163px;"></td>
		</tr>
		<tr height="22">
			<td height="22" style="height:22px;width:172px;">plate holder</td>
			<td style="width:151px;">Viexen, Co.,Ltd.</td>
			<td style="width:131px;">plate holder SX</td>
			<td style="width:163px;"></td>
		</tr>
		<tr height="43">
			<td height="43" style="height:43px;width:172px;">EOS Kiss X7i&#160;</td>
			<td style="width:151px;">Canon Inc., Japan</td>
			<td style="width:131px;">8594B001</td>
			<td style="width:163px;">with a standard zoom lens,&#160; EFP 18-55 mm</td>
		</tr>
		<tr height="22">
			<td height="22" style="height:22px;width:172px;">photographic software</td>
			<td style="width:151px;">Canon Inc., Japan</td>
			<td style="width:131px;">EOS Utility</td>
			<td style="width:163px;"></td>
		</tr>
		<tr height="22">
			<td height="22" style="height:22px;width:172px;">PC</td>
			<td style="width:151px;">Microsoft</td>
			<td style="width:131px;">Surface</td>
			<td style="width:163px;"></td>
		</tr>
		<tr height="22">
			<td height="22" style="height:22px;width:172px;">polarization microscope</td>
			<td style="width:151px;">Olympus</td>
			<td style="width:131px;">BX51</td>
			<td style="width:163px;"></td>
		</tr>
		<tr height="43">
			<td height="43" style="height:43px;width:172px;">first order retardation plate</td>
			<td style="width:151px;">Olympus</td>
			<td style="width:131px;">U-TP530</td>
			<td style="width:163px;">&#955; = 530 nm</td>
		</tr>
		<tr height="22">
			<td height="22" style="height:22px;width:172px;">CCD camera</td>
			<td style="width:151px;">Olympus</td>
			<td style="width:131px;">DP80</td>
			<td style="width:163px;"></td>
		</tr>
		<tr height="22">
			<td height="22" style="height:22px;width:172px;">photographic software</td>
			<td style="width:151px;">Olympus</td>
			<td style="width:131px;">cellSens Standard</td>
			<td style="width:163px;"></td>
		</tr>
		<tr height="43">
			<td height="43" style="height:43px;width:172px;">Java-based image processing program</td>
			<td style="width:151px;">the National Institutes of Health</td>
			<td style="width:131px;">ImageJ</td>
			<td style="width:163px;"></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?...

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 ...

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Bridging the Bio-Electronic Interface with Biofabrication

生物电子界面衔接与Biofabrication

Advancements in lab-on-a-chip technology promise to revolutionize both research and medicine through lower costs, better sensitivity, portability, and ...

科研

生物工程

Flexural Rigidity Measurements of Biopolymers Using Gliding Assays

使用滑翔分析生物大分子的抗弯刚度测量

Microtubules are cytoskeletal polymers which  play a role in cell division, cell mechanics, and intracellular transport. Each of these functions requires ...

Cell Patterning on Photolithographically Defined 
Parylene-C: SiO2 Substrates

Cell Patterning on Photolithographically Defined 
Parylene-C: SiO2 Substrates

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

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

Molecular Entanglement and Electrospinnability of Biopolymers

分子纠缠与生物大分子的Electrospinnability

Electrospinning is a fascinating technique to fabricate micro- to nano-scale fibers from a wide variety of materials. For biopolymers, molecular ...

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

基因转染向着文化微模式球体板细胞基因修饰的细胞移植

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

Methods for Characterizing the Co-development of Biofilm and Habitat Heterogeneity

方法表征的共同发展生物膜和生境异质性

Biofilms are surface-attached microbial communities that have complex structures and produce significant spatial heterogeneities. Biofilm development is ...

PIP-on-a-chip: A Label-free Study of Protein-phosphoinositide Interactions

PIP-on-a-chip：蛋白质 - 磷酸肌醇相互作用的无标记研究

Numerous cellular proteins interact with membrane surfaces to affect essential cellular processes. These interactions can be directed towards a specific ...

Microhoneycomb Monoliths Prepared by the Unidirectional Freeze-drying of Cellulose Nanofiber Based Sols: Method and Extensions

纤维素碳纤维溶胶单向冷冻干燥制备 Microhoneycomb 石柱: 方法和外延

Monolithic honeycomb structures have been attractive to multidisciplinary fields due to their high strength-to-weight ratio. Particularly, microhoneycomb ...

Comprehensive Evaluation of the Effectiveness and Safety of Placenta-Targeted Drug Delivery Using Three Complementary Methods

用三种互补方法综合评价胎盘靶向药物的有效性和安全性

No effective treatments currently exist for placenta-associated pregnancy complications, and developing strategies for the targeted delivery of drugs to ...

Methods for In Vivo Biomechanical Testing on Brachial Plexus in Neonatal Piglets

Methods for In Vivo Biomechanical Testing on Brachial Plexus in Neonatal Piglets

新生儿小猪体内胸腺菌的活体力学试验方法

Neonatal brachial plexus palsy (NBPP) is a stretch injury that occurs during the birthing process in nerve complexes located in the neck and shoulder ...