Name: mechanostimulation of multicellular organisms through a high-throughput microfluidic compression system
Uploaded: 2022-12-23T13:15:00
Description: Watch this Scientific Journal Video about Mechanostimulation of Multicellular Organisms Through a High-Throughput Microfluidic Compression System at JoVE.com

这项工作得到了美国国家科学基金会（CMMI-1946456）、空军科学研究办公室（FA9550-18-1-0262）和美国国立卫生研究院（R01AG06100501A1;R21AR08105201A1）。

1. 硅片模具的制备
<ol><li>首先用丙酮清洁硅片（见 材料表），然后用异丙醇（IPA）清洁。</li>
	<li>将硅晶片放在250°C热板上30分钟以进行脱水烘烤（图3A）。</li>
	<li>在蒸汽底炉中用六甲基二硅氮烷（HDMS）涂覆硅晶片（见 材料表）（工艺温度：150°C，工艺压力：2托，工艺时间：5分钟，HDMS体积：5μL）（图3B）。</l...

<ol>
	<li>Wang, J. H. -. C., Thampatty, B. P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=An+introductory+review+of+cell+mechanobiology.">An introductory review of cell mechanobiology.</a> Biomechanics and Modeling in Mechanobiology. 5 (1), 1-16 (2006).</li><li>Ingber, D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Mechanobiology+and+diseases+of+mechanotransduction.">Mechanobiology and diseases of mechanotransduction.</a> Annals of Medicine. 35 (8), 564-577 (2003).</li><li>Nims, R. J., Pferdehirt, L., Guilak, F. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Mechanogenetics:+Harnessing+mechanobiology+for+cellular+engineering.">Mechanogenetics: Harnessing mechanobiology for cellular engineering.</a> Current Opinion in Biotechnology. 73, 374-379 (2022).</li><li>Bellin, R. M., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Defining+the+Role+of+Syndecan-4+in+Mechanotransduction+using+Surface-Modification+Approaches.">Defining the Role of Syndecan-4 in Mechanotransduction using Surface-Modification Approaches.</a> Proceedings of the National Academy of Sciences. 106, 22102-22107 (2009).</li><li>Simpson, L. J., Reader, J. S., Tzima, E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Mechanical+regulation+of+protein+translation+in+the+cardiovascular+system.">Mechanical regulation of protein translation in the cardiovascular system.</a> Frontiers in Cell and Developmental Biology. 8, 34 (2020).</li><li>Humphrey, J. D., Schwartz, M. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Vascular+mechanobiology:+Homeostasis,+adaptation,+and+disease.">Vascular mechanobiology: Homeostasis, adaptation, and disease.</a> Annual Review of Biomedical Engineering. 23, 1-27 (2021).</li><li>Maurer, M., Lammerding, 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+driving+force:+Nuclear+mechanotransduction+in+cellular+function,+fate,+and+disease.">The driving force: Nuclear mechanotransduction in cellular function, fate, and disease.</a> Annual Review of Biomedical Engineering. 21, 443-468 (2019).</li><li>Jennings, B. H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Drosophila-A+versatile+model+in+biology+&amp;+medicine.">Drosophila-A versatile model in biology &amp; medicine.</a> Materials Today. 14 (5), 190-195 (2011).</li><li>Konno, M., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=State-of-the-art+technology+of+model+organisms+for+current+human+medicine.">State-of-the-art technology of model organisms for current human medicine.</a> Diagnostics. 10 (6), 392 (2020).</li><li>Morgan, T. H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Sex+limited+inheritance+in+Drosophila.">Sex limited inheritance in Drosophila.</a> Science. 32 (812), 120-122 (1910).</li><li>Wu, Q., Kumar, N., Velagala, V., Zartman, J. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Tools+to+reverse-engineer+multicellular+systems:+Case+studies+using+the+fruit+fly.">Tools to reverse-engineer multicellular systems: Case studies using the fruit fly.</a> Journal of Biological Engineering. 13 (1), 1-16 (2019).</li><li>Jayamohan, H., et al., Patrinos, G., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Chapter+11+-+Advances+in+Microfluidics+and+Lab-on-a-Chip+Technologies.">Chapter 11 - Advances in Microfluidics and Lab-on-a-Chip Technologies.</a> Molecular Diagnostics. , 197-217 (2017).</li><li>Scheler, O., Postek, W., Garstecki, P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Recent+developments+of+microfluidics+as+a+tool+for+biotechnology+and+microbiology.">Recent developments of microfluidics as a tool for biotechnology and microbiology.</a> Current Opinion in Biotechnology. 55, 60-67 (2019).</li><li>Mohammed, D., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Innovative+tools+for+mechanobiology:+Unraveling+outside-in+and+inside-out+mechanotransduction.">Innovative tools for mechanobiology: Unraveling outside-in and inside-out mechanotransduction.</a> Frontiers in Bioengineering and Biotechnology. 7, 162 (2019).</li><li>Shorr, A. Z., S&#246;nmez, U. M., Minden, J. S., LeDuc, P. R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=High-throughput+mechanotransduction+in+Drosophila+embryos+with+mesofluidics.">High-throughput mechanotransduction in Drosophila embryos with mesofluidics.</a> Lab on a Chip. 19 (7), 1141-1152 (2019).</li><li>Kim, Y. T., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Mechanochemical+Actuators+of+Embryonic+Epithelial+Contractility.">Mechanochemical Actuators of Embryonic Epithelial Contractility.</a> Proceedings of the National Academy of Sciences. 40, 14366-14371 (2014).</li><li>Ashburner, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> Drosophila. A Laboratory Handbook. , (1989).</li><li>Qin, D., Xia, Y., Whitesides, G. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Soft+lithography+for+micro-and+nanoscale+patterning.">Soft lithography for micro-and nanoscale patterning.</a> Nature Protocols. 5 (3), 491 (2010).</li><li>Lee, H., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+new+fabrication+process+for+uniform+SU-8+thick+photoresist+structures+by+simultaneously+removing+edge+bead+and+air+bubbles.">A new fabrication process for uniform SU-8 thick photoresist structures by simultaneously removing edge bead and air bubbles.</a> Journal of Micromechanics and Microengineering. 21 (12), 125006 (2011).</li><li>Xia, Y., Whitesides, G. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Soft+lithography.">Soft lithography.</a> Annual Review of Materials Science. 28 (1), 153-184 (1998).</li><li>Sonmez, U. M., Coyle, S., Taylor, R. E., LeDuc, P. R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Polycarbonate+heat+molding+for+soft+lithography.">Polycarbonate heat molding for soft lithography.</a> Small. 16 (16), 2000241 (2020).</li><li>Levario, T. J., Zhan, M., Lim, B., Shvartsman, S. Y., Lu, H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Microfluidic+trap+array+for+massively+parallel+imaging+of+Drosophila+embryos.">Microfluidic trap array for massively parallel imaging of Drosophila embryos.</a> Nature Protocols. 8 (4), 721-736 (2013).</li></ol>

本文介绍了一种微流体装置的开发，该装置可以自动对齐，固定并精确地将机械刺激应用于数百个完整的 果蝇 胚胎。微流体系统与薄玻璃盖玻片的集成允许在刺激期间用高分辨率共聚焦显微镜对胚胎进行成像。微流体装置还能够在刺激后立即收集胚胎以进行下游生物测定。详细描述了该器件的设计考虑因素、制造方法和特性。硅晶圆模具制造方案允许光刻胶具有高纵横比沟槽的均匀厚涂?...

生命系统在其一生中不断经历并响应各种机械输入1.机械转导与许多疾病有关，包括发育障碍、肌肉和骨质流失以及通过直接或间接受机械环境影响的信号通路的神经病理学2。然而，机械敏感信号通路4中受机械刺激3调控的基因和蛋白质在很大程度上仍然未知5，阻碍了机械调节机制的阐明和与病理机械转导相关的疾病的分子靶点的鉴定6，7.将机械生物学研究投射到相关生理过程的一个限制因素是使用具有常规培养皿的单个细胞而不是完整的多细胞生物。模式生物，如黑腹果蝇（果蝇），对理解参与动物发育的基因、信号通路和蛋白质做出了巨大贡献8，9，10。然而，在机械生物学研究中使用果蝇和其他多细胞模式生物受到实验工具挑战的阻碍。用于准备、分类、成像或应用各种刺激的传统技术大多需要手动操作;这些方法非常耗时，需要专业知识，引入可变性，并限制实验设计和样本量11。最近的微技术进步是实现具有非常高通量和高度受控实验参数的新型生物测定的重要资源12，13，14。
本文介绍了一种增强型微流体装置的开发，该装置以单轴压缩的形式对准、固定和精确地将机械刺激应用于数百个完整的 果蝇 胚胎15（图 1）。微流体系统与玻璃盖玻片的集成允许在刺激期间对样品进行高分辨率共聚焦成像。微流体装置还能够在刺激后快速收集胚胎以进行组学测定（图2）。本文描述了该装置的设计考虑因素，以及使用软光刻和实验表征的制造。由于制造这种器件的硅晶圆模具需要在具有高纵横比（AR）沟槽（AR >5）的大面积上均匀涂覆厚光刻胶（厚度>200μm），因此该方法大大修改了传统的光刻模具制造方案。通过这种方式，该方法促进了光刻胶的处理、粘附、涂覆、图案化和开发。此外，还讨论了潜在的陷阱及其解决方案。最后，使用其他多细胞系统（如 果蝇 卵室和脑类器官16）证明了这种设计和制造策略的多功能性。

<table border="1" fo:keep-together.within-page="1" fo:keep-with-next.within-page="always">
	<tbody>
		<tr>
			<td>100 mL tri-cornered perforated plastic beakers with 60 mm Petri dishes</td>
			<td>Fisher</td>
			<td>14-955-111B</td>
			<td>Perferate with air holes</td>
		</tr>
		<tr>
			<td>100 mm P B &#60;100&#62; 0-100 500um SSP Test Grade Si Wafer</td>
			<td>University Wafer</td>
			<td>452</td>
			<td></td>
		</tr>
		<tr>
			<td>Biopsy punches</td>
			<td>Ted Pella</td>
			<td>15110</td>
			<td></td>
		</tr>
		<tr>
			<td>Bleach</td>
			<td></td>
			<td></td>
			<td>Not brand specific</td>
		</tr>
		<tr>
			<td>Blunt needle set</td>
			<td>CML Supply</td>
			<td>901</td>
			<td></td>
		</tr>
		<tr>
			<td>Contact Mask Aligner</td>
			<td>Quintel</td>
			<td>Q4000 MA</td>
			<td></td>
		</tr>
		<tr>
			<td>Cutting mat</td>
			<td>Dahle</td>
			<td>Vantage 10670</td>
			<td>size: 24&#34; x 36&#34;</td>
		</tr>
		<tr>
			<td>Developer</td>
			<td>Kayaku Advance Materials</td>
			<td>SU-8 2000</td>
			<td></td>
		</tr>
		<tr>
			<td>Direct Write Lithographer</td>
			<td>Heidelberg</td>
			<td>MLA100</td>
			<td></td>
		</tr>
		<tr>
			<td>Dissecting microscope</td>
			<td></td>
			<td></td>
			<td>Any commericailly availble dissecting microscope with transmitted light</td>
		</tr>
		<tr>
			<td>Glass petri dish</td>
			<td>Fisher</td>
			<td>FB0875713A</td>
			<td></td>
		</tr>
		<tr>
			<td>Glass slide</td>
			<td>Warner Instruments</td>
			<td>64-0710&#160; (CS-24/60)</td>
			<td></td>
		</tr>
		<tr>
			<td>HMDS Vapor Prime Oven</td>
			<td>Yes Engineering</td>
			<td>YES-3TA</td>
			<td></td>
		</tr>
		<tr>
			<td>NaCl</td>
			<td></td>
			<td></td>
			<td>Not brand specific</td>
		</tr>
		<tr>
			<td>Oven</td>
			<td>Labnet</td>
			<td>I5110A</td>
			<td></td>
		</tr>
		<tr>
			<td>Paintbrush</td>
			<td></td>
			<td></td>
			<td>Not brand specific</td>
		</tr>
		<tr>
			<td>PDMS</td>
			<td>Dow Corning</td>
			<td>Sylgard 184</td>
			<td></td>
		</tr>
		<tr>
			<td>Photoresist</td>
			<td>MicroChem</td>
			<td>SU-8 2100</td>
			<td></td>
		</tr>
		<tr>
			<td>Plasma cleaner</td>
			<td>Harrick Plasma</td>
			<td>PDC-32G</td>
			<td></td>
		</tr>
		<tr>
			<td>Portable pressure source</td>
			<td>hygger Quietest</td>
			<td>HGD946</td>
			<td></td>
		</tr>
		<tr>
			<td>Pressure gauge</td>
			<td>Cole-Parmer</td>
			<td>EW-68950-25</td>
			<td></td>
		</tr>
		<tr>
			<td>Spin Coater</td>
			<td>Laurell</td>
			<td>WS-650-8B</td>
			<td></td>
		</tr>
		<tr>
			<td>Trichloro(1H,1H,2H,2H-perfluorooctyl)silane (PFOCTS)</td>
			<td>Sigma-Aldrich</td>
			<td>448931-10G</td>
			<td></td>
		</tr>
		<tr>
			<td>Triton-X 100</td>
			<td>Fisher</td>
			<td>AAA16046AP</td>
			<td></td>
		</tr>
		<tr>
			<td>Tubing</td>
			<td>Saint-Gobain</td>
			<td>02-587-1A</td>
			<td></td>
		</tr>
		<tr>
			<td>Ultrasonic Cleaner</td>
			<td>Cole-Parmer</td>
			<td>UX-08895-05</td>
			<td></td>
		</tr>
		<tr>
			<td>Vacuum Pump</td>
			<td>Cole-Parmer</td>
			<td>EW-07164-87</td>
			<td></td>
		</tr>
	</tbody>
</table>

mechanostimulation of multicellular organisms through a high-throughput microfluidic compression system

在胚胎发生过程中，协调的细胞运动产生调节基因表达和活性的机械力。为了研究这一过程，已经使用抽吸或盖玻片压缩等工具来机械刺激整个胚胎。这些方法限制了实验设计，因为它们不精确，需要人工处理，并且只能同时处理几个胚胎。微流体系统在自动化此类实验任务的同时提高通量和精度方面具有巨大潜力。本文描述了一种微流体系统，用于精确压缩整个 黑腹果 蝇（果蝇）胚胎。该系统具有带有气动驱动可变形侧壁的微通道，可实现胚胎对齐、固定、压缩和刺激后收集。通过将这些微通道并行到七个通道中，可以同时将稳定或动态压缩模式应用于数百个 果蝇 胚胎。在玻璃盖玻片上制造该系统有助于使用高分辨率显微镜同时对样品进行机械刺激和成像。此外，利用生物相容性材料（如PDMS）以及流体流过系统的能力使该设备能够对介质依赖性样品进行长期实验。这种方法还消除了手动安装的需要，手动安装会对样品施加机械压力。此外，从微通道快速收集样品的能力使刺激后分析成为可能，包括需要大量样品的组学测定，这是使用传统机械刺激方法无法实现的。该系统的几何形状易于扩展到不同的生物系统，使许多领域能够从本文描述的功能特征中受益，包括高样品通量、机械刺激或固定以及自动比对。

微流体系统分为两个由可变形的PDMS侧壁隔开的子隔间。第一个隔间是液体系统，其中果 蝇 胚胎被引入，自动对齐，排列和压缩。第二个隔室是气体系统，其中压缩通道两侧的气体压力通过死端微通道 控制 ，以精确控制压缩通道的有效宽度。微流体装置在底部用载玻片密封，从而能够对微流控装置相关尺寸的样品进行高分辨率实时成像（补充图2）。
<p class="jove_conten...

Watch this Scientific Journal Video about Mechanostimulation of Multicellular Organisms Through a High-Throughput Microfluidic Compression System at JoVE.com

Mechanostimulation of Multicellular Organisms Through a High-Throughput Microfluidic Compression System

本协议描述了微流体系统的设计，制造和表征，该系统能够以最少的用户干预对齐，固定和精确压缩数百个 黑腹果蝇 胚胎。该系统可实现高分辨率成像和样品回收，以进行刺激后分析，并且可以扩展以适应其他多细胞生物系统。

通过高通量微流体压缩系统对多细胞生物进行机械刺激

During embryogenesis, coordinated cell movement generates mechanical forces that regulate gene expression and activity. To study this process, tools such ...

该协议解释了如何制造和使用微流体技术，以利用样品通量，自动处理以及精确施加机械应力的能力，同时可视化样品。该技术最大限度地减少了对多细胞生物有机体（如胚胎和类器官）的手动处理，以进行固定、比对和实时成像。它还能够将机械压缩应用于机械生物学研究。使用传统的微加工技术，为这种微流控芯片制造具有高纵横比特征的模具可能具有挑战性。本视频文章中介绍的技巧可以克服这些挑战。首先，首先用丙酮清洁硅片，然后用异丙醇清洁。将硅晶片放在 250 摄氏度的热板上 30 分钟。用于脱水烘烤。在蒸汽主炉中用六甲基二硅氧烷涂覆硅片。将一瓶SU-8 2100光刻胶放入60摄氏度的烤箱中15分钟以降低其粘度。将一毫升加热的光刻胶倒入放置在热板上的每一英寸硅晶片，直到光刻胶覆盖大部分表面。首先以每分钟 250 转的速度应用预旋转，持续 30 秒，然后以每分钟 350 转的速度再应用预旋转，持续 30 秒，两者的加速度均为每秒 100 RPM。接下来，首先以每分钟 500 转的速度应用旋转，持续 15 秒，每秒加速度为 100 RPM，然后以每秒 1， 450 转的速度旋转 30 秒，加速度为 300 RPM。用洁净室拭子取出边缘珠，并在晶圆上喷洒丙酮以去除瑕疵并促进均匀涂层。通过光掩模将硅胶晶圆暴露在每平方厘米350毫焦耳的紫外线下。使用接触掩模对准器，将曝光后烘烤应用于硅晶圆，并使其冷却至室温。将烧杯放入另一个较大的烧杯中，并在较大的烧杯中填充新鲜的显影剂溶液。将金属网放在烧杯上。将硅晶片倒置放在金属网上。在搅拌器打开的情况下，将硅晶片浸没在显影剂中 30 分钟。将硅晶片转移到装有新鲜显影剂的超声波浴超声仪中，以 40 千赫兹的速度进行一小时。然后从烧杯中取出晶圆，并用新鲜的显影液清洗。通过将PDMS碱与固化剂以10：1的比例混合来制备预固化的PDMS溶液。通过将混合物放入室温下500g的离心机中5分钟来脱气。将预固化的PDMS倒在硅晶片上，然后再次脱气。最后，将未固化的PDMS放入60摄氏度的烤箱中固化。使用手术刀切割与微流控芯片几何形状相对应的固化PDMS区域的边界。使用活检打孔器或带有钝尖的针头在 PDMS 上打孔。等离子处理后，将PDMS放在载玻片上，其图案表面朝向载玻片，通过共价键密封微通道。允许俄勒冈-R成年苍蝇在苹果汁琼脂平板上产卵，并在给定实验产卵后的所需发育时间收集平板。用胚胎蛋液淹没琼脂，并用画笔轻轻搅拌胚胎以将它们从琼脂中取出。将胚胎转移到50%漂白剂溶液中90秒，偶尔搅拌。通过组织筛过滤胚胎，并用水彻底洗去漂白剂溶液。将胚胎转移到90毫米玻璃培养皿中，并带有足够的胚胎蛋液以完全覆盖胚胎。在解剖显微镜上用透照检查胚胎，并选择所需发育阶段的胚胎以加载到微流体装置中。通过主胚胎入口向所有七个胚胎微通道填充0.4微米的过滤IPA。用0.4微米的过滤去离子水代替IPA。然后用胚胎蛋液代替去离子水。使用玻璃移液管从玻璃培养皿中收集大约 100 个预选胚胎。接下来，将胚胎移液到胚胎入口，并使用便携式真空泵向气体入口施加大约三个PSI负压以打开胚胎微通道。然后将微流控芯片向下倾斜，使胚胎自动对齐并沉淀到胚胎微通道中。如果胚胎微通道入口被同时进入的多个胚胎堵塞，请将微流控芯片向上倾斜，然后再次向下倾斜以清除堵塞。根据所需的吞吐量，将多达300个胚胎引入胚胎微通道。胚胎加载完成后，移除真空以固定胚胎。然后将微流控芯片倾斜回水平位置。将带有压力表的便携式正压源连接到气体入口，以施加三个PSI压缩。如果要对机械刺激的胚胎进行实时成像实验，请将微流控芯片放在标准显微镜载物台玻璃载玻片支架上，气体入口连接到压力源。在微流体通道内的荧光显微镜下检查胚胎。压缩实验完成后，可以收集胚胎进行下游分析。为此，首先，将真空施加到进气口以释放胚胎。然后将微流控芯片向上倾斜，使胚胎向下移动到胚胎引入口。使用玻璃移液管从微流控芯片中收集胚胎。通过将果蝇胚胎加载到压缩通道中并向气体通道施加正压来实验确定微流体装置的功能。胚胎在真空下或中性压力状态下不会经历显着的压缩。当施加正压时，它们被压缩。在显微镜下测量胚胎宽度减小的尺寸证明了如何使用气体压力来获得目标压缩水平。以这种方式制造的微流体装置允许对固定的样品进行化学刺激。刺激允许高时空成像。发育生物学的一个基本问题是机械力如何在发育过程中调节蛋白质和基因表达？这项技术使我们能够一次对大量胚胎进行机械刺激，以便我们可以分离出足够量的蛋白质和RNA来进行比较蛋白质组学或转录组学实验。这将使研究人员能够更多地了解对机械力敏感的蛋白质和基因。

Research

JoVE Journal

Bioengineering

Polymer Microarrays for High Throughput Discovery of Biomaterials

聚合物芯片高通量发现的生物材料

The discovery of novel biomaterials that are optimized for a specific biological application is readily achieved using polymer microarrays, which allows a ...

科研

生物工程

High-throughput Protein Expression Generator Using a Microfluidic Platform

利用微流控平台的高通量蛋白质表达发生器

Rapidly increasing fields, such as systems biology, require the development and implementation of new technologies, enabling high-throughput and ...

High Throughput Microfluidic Rapid and Low Cost Prototyping Packaging Methods

In this work, 3 different packaging and assembly techniques are presented. They can be classified into two categories: one-time use and reusable packaging ...

Systematic Analysis of In Vitro Cell Rolling Using a Multi-well Plate Microfluidic System

Systematic Analysis of In Vitro Cell Rolling Using a Multi-well Plate Microfluidic System

的系统分析<em&gt;体外</em&gt;细胞滚动使用多孔板微流控系统

A major challenge for cell-based therapy is the inability to systemically target a large quantity of viable cells with high efficiency to tissues of ...

Use of a High-throughput In Vitro Microfluidic System to Develop Oral Multi-species Biofilms

Use of a High-throughput In Vitro Microfluidic System to Develop Oral Multi-species Biofilms

使用高通量的<em&gt;在体外</em&gt;微流控系统开发口服多物种生物膜

There are few high-throughput in vitro systems which facilitate the development of multi-species biofilms that contain numerous species commonly detected ...

Using Tomoauto: A Protocol for High-throughput Automated Cryo-electron Tomography

使用Tomoauto：一个协议，用于高通量自动化低温电子断层扫描

Cryo-electron tomography (Cryo-ET) is a powerful three-dimensional (3-D) imaging technique for visualizing macromolecular complexes in their native ...

A Microfluidic Platform for High-throughput Single-cell Isolation and Culture

高通量单细胞的分离和文化的微流体平台

Studying the heterogeneity of single cells is crucial for many biological questions, but is technically difficult. Thus, there is a need for a simple, yet ...

High-throughput Identification of Bacteria Repellent Polymers for Medical Devices

医疗器械细菌高通量鉴定驱蚊聚合物

Medical devices are often associated with hospital-acquired infections, which place enormous strain on patients and the healthcare system as well as ...

Use of High-Throughput Automated Microbioreactor System for Production of Model IgG1 in CHO Cells

高通量自动 Microbioreactor 系统在 CHO 细胞模型 IgG1 生产中的应用

Automated microscale bioreactors (15 mL) can be a useful tool for cell culture engineers. They facilitate the simultaneous execution of a wide variety of ...

A Microfluidic Platform for Stimulating Chondrocytes with Dynamic Compression

微流体平台，用于用动态压缩刺激软骨细胞

Mechanical stimuli are known to modulate biological functions of cells and tissues. Recent studies have suggested that compressive stress alters growth ...