这项工作得到了犹他大学副会长办公室的支持, 以及与 P30 CA042014 授予洪博培癌症研究所的拨款以及洪博培癌症研究所的 CRR 计划。

1. 烷阴性的制作
<ol><li>
烷的制备
	<ol>
<li>测量18克的有机硅弹性体和1.8 克的固化试剂。将固化试剂倒入含有弹性体的测量船上。</li>
		<li>将弹性体和固化试剂搅拌1分钟, 将混合物放入真空室30分钟。</li>
		<li>从真空中取出该硅烷, 用一个曲奇刀 (半径 = 3.8 厘米) 将 15 g 倒到底片上, 以防止其从侧面跑掉, 并覆盖其余的混合材料。</li>
		<li>将含有该硅烷的模具放入真空室10分钟。</li>
		<li>从真空室取出含有该硅烷的模具, 将其放在热板上, 设置为150°c 60 分钟即可固化, 并用铝箔盖住。</li>
		<li>将剩余的新的一级玻璃培养皿 (直径为10厘米) 倒入另一台, 用另一个曲奇刀作为模具, 创建一垫。把这个放进真空室, 直到它的阴性完全固化。</li>
		<li>将培养皿从真空转到预热热板, 在150摄氏度处进行60分钟的固化。</li>
	</ol>
</li>
	<li>
烷的等离子表面处理
	<ol>
<li>插入一个锋利的剃刀刀片下的, 并从饼干切割机, 以谨慎地删除从负面的。</li>
		<li>用一把锋利的嗜好刀, 从更大的部分中切下底片;它应该被削减与足够的缓冲面积, 以附加另一块在它的顶部, 而不妨碍负面。</li>
		<li>插入一个锋利的剃刀刀片下面的固化与阴性, 以谨慎地从培养皿中取出。</li>
		<li>使用一个爱好刀切出一张从垫子上的一块, 这是相同大小的一块切的底片。然后, 用足够的空间在微通道上剪下一个长方形, 当它放在负极的顶端时, 它就能适应它。</li>
		<li>将两片 (负极和矩形) 放在平坦的基板上 (如由玻璃、石英、聚苯乙烯或其他材料制成的幻灯片), 并将其插入射频 (RF) 等离子清洗器中。关上车门, 用真空泵把空气排出, 封住房间。使用数字真空压力控制器将空气注入 400 mTorr 的压力。 注: 如果用于等离子清洗的基板未在等离子清洁器中使用, 则在处理阴性之前将其放入等离子清洗剂中, 以防止丙烯酸基层粘附到基体上。</li>
		<li>将 RF 设置变高, 并在 "查看" 窗口中显示粉红色的空气等离子。等离子-在三十年代将这两片微波治疗, 然后将射频设置关闭。允许空气缓慢进入会议厅, 以防止湍流气流干扰会议厅的内容。</li>
		<li>从等离子清洗器中取出该台式, 并将其放在一个。然后, 小心地将该矩形的反相反转到底片上, 然后用一对镊子把它压下来。让它休息10分钟, 让这两个部分, 以坚持彼此。</li>
	</ol>
</li>
	<li>
Fluorosilane 表面处理烷阴性
	<ol>
<li>用吸管把2滴氯 (1 小时, 1 小时, 2 小时, 2 氟辛烷) 硅 (PFOTS) 成小重量的船, 并插入玻璃真空室。</li>
		<li>将其反转到一块铝箔上 (使微通道负极朝上) 并将其放入玻璃真空室。然后, 连续疏散房间24小时。</li>
	</ol>
</li>
</ol>2. MY133 微通道的制作
<ol><li>
MY133-V2000 微通道结构的形成
	<ol>
<li>用400µL 的 MY133-V2000 来填充。 注: 阴性必须略溢出。</li>
		<li>将 MY133-V2000-filled 的负极注入真空室2小时, 以除去任何气泡。</li>
		<li>从真空中取出 MY133-V2000, 然后慢慢地按下溢出的顶部的玻璃滑动, 创建 MY133-V2000 的平坦表面, 防止氧气抑制聚合。 注: 在通道表面的聚硅烷, 会部分抑制聚合, 从而在以后的阶段进行粘结。与石英相比, 玻璃滑块的 UV 透明度略有降低 (35%), 这有助于防止固化装置的任何发黄。</li>
		<li>将 MY133-V2000 插入 400 W uv 烤箱, 并将紫外线辐射量设置为300s 最大强度的 50%, 以治疗 MY133-V2000 微通道。 注: 用于治疗该装置的峰值波长约为375毫微米, 带宽约为 25 nm。大约4500兆焦耳/cm2的功率被用来治疗微通道, 这是稍微超过双倍的最低固化功率的制造商建议。</li>
	</ol>
</li>
	<li>
建立丙烯酸支架
	<ol>
<li>使用矢量绘图软件绘制丙烯酸基层。确保基层为1.5 毫米厚、75毫米长和25毫米宽的矩形, 其中心矩形测量25毫米 x 11 毫米。</li>
		<li>使用矢量绘图软件绘制丙烯酸中层。确保中间层为矩形 (1.5 毫米厚, 75 毫米长, 25 毫米宽), 中心正方形 (5 毫米 x 5 毫米) 和两个圆 (直径为3毫米), 用15毫米隔开。</li>
		<li>使用矢量绘图软件绘制丙烯酸顶层。确保顶层是矩形 (3 毫米厚, 30 毫米长, 25 毫米宽), 中心正方形 (5 毫米 x 5 毫米) 和两个圆圈 (两个直径为3毫米), 用15毫米隔开。</li>
		<li>剪掉在矢量绘图软件中用激光切割器创建的设备设计。使用100% 电源和30% 速度的示例设置。运行该程序两次, 以确保丙烯酸被切断。</li>
		<li>使用尺寸 M5 0.80 mm 攻丝工具, 点击丙烯酸顶层的孔。</li>
		<li>用丙酮擦拭丙烯酸片, 除去残留痕迹或灼伤。</li>
	</ol>
</li>
	<li>
MY133-V2000 在丙烯酸酯支架上的粘结
	<ol>
<li>使用粘合剂 (如氰超级胶水) 将丙烯酸的基层附着在玻璃基板上。在丙烯酸的边缘放置两个小水滴, 然后使用一次性工具均匀地分散胶水。将玻璃基板放在丙烯酸上, 并允许胶水用玻璃的重量将其保持在原处。</li>
		<li>用正排量吸管将所暴露的玻璃涂上100µL 的硅烷;然后, 将基层插入真空旋转涂布机, 并将其设置为 1500 rpm, 以2分钟均匀地将玻璃涂上大约 10-µm 厚的一部。</li>
		<li>从旋转涂布机中取下基层, 并仔细擦拭所有用丙酮和纸巾涂丙烯酸的多余的塑料。小心不要打扰成灾。</li>
		<li>用65摄氏度的烤箱将基层烘烤为2小时, 以治疗该硅烷。</li>
		<li>在玻璃滑梯上用1毫升的硅烷, 然后, 再用另一张玻璃滑梯均匀地将其分散, 直到它开始渗出两侧。在150摄氏度的热板上固化10分钟。</li>
		<li>将固化的 MY133-V2000 切割成一个与该设备尺寸相同的矩形, 然后使用丙烯酸的中间层作为模具, 为储层打孔, 并在其上切割一个方形查看窗口, 使其成为塑料垫片。</li>
		<li>将 MY133-V2000, 通道侧向上, 在同一个或类似的平坦基底上, 用于等离子处理在步骤1.2.5 中的。将丙烯酸基层放置在 O2等离子清洗器的 MY133-V2000 通道旁边。将真空压力设置为 200 mTorr, 射频电平高, 表面处理三十年代的通道和玻璃基板。</li>
		<li>使用镊子, 立即将 MY133-V2000 通道侧向下放置在基层丙烯酸的矩形切口中, 这样 MY133-V2000 就能接触到该硅烷。</li>
		<li>在 MY133-V2000 设备的顶部放置一个, 在垫片上的孔与装置中的储层衬在一起。</li>
		<li>将未完成的设备放到工作台上方凸起的平台上, 以提供设备装配的夹紧面。</li>
		<li>用注射器提取3毫升丙烯酸水泥。在基层顶部分配足够的丙烯酸水泥, 以提供薄薄的涂层。尽量使大衣尽可能, 不要让材料渗入通道。</li>
		<li>将中层丙烯酸片放在基层丙烯酸的顶部。确保孔与 MY133-V2000 通道的储层一致。</li>
		<li>用3小钳夹住基层和中间层, 尽可能紧固2分钟, 而丙烯酸水泥硬化, 将丙烯酸片粘合在一起。确保在这一步骤中, 水库不会受阻, 以防止空气被困在 MY133-V2000 装置下面。</li>
		<li>从设备上卸下压力, 将丙烯酸水泥放在与丙烯酸顶层预期接触部位的中间层。</li>
		<li>将丙烯酸的顶层放在丙烯酸的中间层片上, 确保孔与储层对齐, 并允许它在工作台上休息2分钟, 而丙烯酸水泥干燥。</li>
	</ol>
</li>
</ol>3. MY133-V2000 装置的测试和使用
<ol><li>
泄漏测试
	<ol>
<li>通过将10µL 的食品染料或去离子水添加到一个储孔中, 测试 MY133-V2000 装置的粘附和流动。</li>
		<li>在储层插入一根管子 (直径1/8 英寸), 并将另一端连接到真空陷阱。打开真空, 将染料或水从通道中拔出, 以确保创建成功的设备。</li>
		<li>在显微镜下检查, 以验证通道或水库没有渗漏, 用乙醇填充水库, 并允许乙醇在室温下坐10分钟。然后, 通过使用真空来拉动乙醇, 从通道中清洗染料或水。</li>
		<li>用乙醇喷洒通道, 放入无菌聚苯乙烯盘中。与 parafilm 紧密包装, 并将其储存在不育的环境中, 直到需要为止。 注意: 在这一点上, 设备必须处理灌装, 以防止污染。当准备使用该设备时, 容器外部应使用乙醇进行灭菌, 并在打开容器之前将其带入安全柜。</li>
	</ol>
</li>
	<li>
MY133-V2000 装置中的电镀 MCF7 细胞
	<ol>
<li>生长 MCF7 细胞在 10 cm 培养皿在一个完整的媒体组成的 Dulbecco 的最低基本培养基 (DMEM), 补充10% 胎牛血清, 青霉素-链霉素, 谷氨酰胺和非必需氨基酸, 在细胞培养孵化器,保持潮湿的气氛中含有 5% CO2和 20% O2在温度为37摄氏度。在微通道中播种之前, 将细胞生长到大约80% 汇合处。</li>
		<li>首先, 喷洒含有70% 乙醇的微通道的聚苯乙烯盘。然后, 将其放到安全柜中, 然后从环境实验室环境中去除保护通道的 parafilm。</li>
		<li>在微通道表面固定人血浆衍生纤维连接蛋白, 将其在 Dulbecco 磷酸盐缓冲盐水 (DPBS) 中稀释至10µg/毫升。然后, 注入20µL 的这个溶液进入通道和孵化它在20°c 45 分钟。</li>
		<li>将20µL 细胞培养培养基注入通道, 将纤维连接蛋白溶液从通道中冲洗出来, 并使其在20摄氏度处孵化10分钟。</li>
		<li>用10毫升的 DPBS 冲洗细胞, 并胰蛋白酶处理1毫升的1x 胰蛋白酶。孵育细胞在37°c 7 分钟, 中和胰蛋白酶与9毫升完整的培养基。</li>
		<li>注入20µL 的细胞进入通道和孵化他们在37°c 45-120 分钟, 以允许附着在基板成像之前。</li>
	</ol>
</li>
</ol>

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S., Popescu, G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Highly+sensitive+quantitative+imaging+for+monitoring+single+cancer+cell+growth+kinetics+and+drug+response.">Highly sensitive quantitative imaging for monitoring single cancer cell growth kinetics and drug response.</a> PLoS ONE. 9 (2), 89000 (2014).</li></ol>

作者没有什么可透露的。

MY133-V2000 可以替代传统的软光刻制造材料, 如新的。以往的研究表明, 具有较高折射率的材料, 例如, 在通道壁附近引入了重要的工件, 这是由于在管道内的制造材料和水溶液之间的折射指数不匹配。13. MY133-V2000 使微流控装置的折射率与生物医学应用中常用的水溶液相匹配。这减少了成像工件时, 结合微流体与先进的显微技术, 提供了明显的优势比传统的微流控材料。该系统使微流?...

微流控技术的发展已使多种新的生物医学技术能够利用微观尺度流的独特物理1,2。这包括在微流控平台上建立的诊断技术, 量化临床相关生物标志物, 包括细胞刚度3、表面标记4和生长5。通过操纵单细胞, 微流控装置也可用于测量生物标志物的异质性, 例如作为恶性肿瘤6的指示器。将微流控应用与显微技术结合在一起的能力进一步增加了这些平台的效用, 允许同时测量多个生物标志物7的设备。
QPM 是一种显微技术, 用于测量光通过时的相移, 并与透明样品内的物质相互作用。通过使用折射率与生物量密度8、9之间的已知关系, 可以从 QPM 测量中计算出单个细胞的质量。先前的研究表明, QPM 能够通过紊乱强度12测量临床相关参数, 如细胞生长10、11和细胞机械性能。当与微流体结合时, QPM 可能被用来测量体外高度控制环境中细胞的行为。QPM 与微流体相结合的主要挑战之一是,通过软光刻13, 大多数聚合物用于构造微流控通道的高折射率。
微流体与各种显微技术相结合所面临的一个重要挑战是器件材料的折射率相对于水的折射率14,15的不匹配。解决这一问题的一种方法是使用低折射率聚合物, 如 CYTOP16或 MY133-V200013。后者是一种氟化紫外 (UV) 固化丙烯酸酯聚合物, 具有类似于水的折射率 (n = 1.33), 并且与软光刻技术兼容, 允许平滑地融入许多已建立的微流控设备制造工作流。这使得 MY133-V2000 不仅适用于微流控设备的制造, 而且还允许它与 QPM 和其他显微方法相结合, 测量细胞在菌落和单细胞尺度上的行为。MY133-V2000 消除了由于相位解缠而产生的工件, 如光通过 water-MY133 接口, 则生成很少的相移。
虽然消除了折射率的不匹配, 但与由氟化聚合物 (如 MY133-V2000) 制造的器件有关的一个主要挑战是低粘附到其他材料, 如玻璃或硅烷。本工作演示了一种利用软光刻技术制备 MY133-V2000 微流控器件的方法。使用 O2等离子处理通道的表面和与定制的丙烯酸固定器结合, 确保该装置附着在基体上, 形成密封通道。该装置适用于细胞培养和 QPM 测量通道内细胞的质量, 在检测活细胞生长和细胞生物量细胞内转运等方面具有重要的应用价值, 两者在诊断中具有临床意义。医学和药物发现。

<table><tbody><tr><td>MY133-V2000</td><td>MY Polymers</td><td>MY133-V2000</td><td></td></tr><tr><td>Sylgard 184</td><td>Ellsworth Adhesives</td><td>184 SIL ELAST KIT 0.5KG</td><td></td></tr><tr><td>Fisher Premium microscope slides</td><td>Fisher Scientific</td><td>12-544-4</td><td></td></tr><tr><td>.118&quot;(3.0mm) x 12&quot; x 12&quot; Acrylic Sheet</td><td>United States Plastic Corp</td><td>44290</td><td></td></tr><tr><td>.060&quot;(1.5mm) x 12&quot; x 12&quot; Acrylic Sheet</td><td>United States Plastic Corp</td><td>44200</td><td></td></tr><tr><td>SCIGRIP 3 Very Fast Set Acrylic Cement</td><td>United States Plastic Corp</td><td>45735</td><td></td></tr><tr><td>Standard Aluminum Foil (.6 mm thick)</td><td>VWR</td><td>89107-726</td><td></td></tr><tr><td>Kim Wipes</td><td>Fisher Scientific</td><td>06-666</td><td></td></tr><tr><td>Insta-Cure+ Super Glue</td><td>Bob Smith Industries</td><td>BSI-109</td><td></td></tr><tr><td>1/8&quot; PVC tubing</td><td>McMaster Carr</td><td>5231K55</td><td></td></tr><tr><td>McCormick Food Coloring</td><td>Target</td><td>13353207</td><td></td></tr><tr><td>X-Acto #1 Precision Knife</td><td>X-Acto</td><td>X3201</td><td></td></tr><tr><td>X-Acto #18 Heavyweight wood chiseling blade</td><td>X-Acto</td><td>X218</td><td></td></tr><tr><td>VWR Razor Blades</td><td>VWR</td><td>55411-055</td><td></td></tr><tr><td>Surface Treated Cell Culture Dishes</td><td>Fisher Scientific</td><td>FBO12922</td><td></td></tr><tr><td>Fibronectin Human Plasma</td><td>Sigma-Aldritch</td><td>F0895-1MG</td><td></td></tr><tr><td>Trypsin-EDTA 10x</td><td>Fisher Scientific</td><td>15-400-054</td><td></td></tr><tr><td>Corning Dulbecco&#039;s Phosphate Buffered Saline</td><td>Fisher Scientific</td><td>MT21030CM</td><td></td></tr><tr><td>Gibco Penicillin-Streptomycin</td><td>Fisher Scientific</td><td>15-140-148</td><td></td></tr><tr><td>HyClone Nonessential Amino Acids 100x</td><td>Fisher Scientific</td><td>SH3023801</td><td></td></tr><tr><td>Fetal Bovine Serum</td><td>Omega Scientific</td><td>FB-12</td><td></td></tr><tr><td>Corning DMEM with L-glutamine and glucose</td><td>Fisher Scientific</td><td>MT10013CV</td><td></td></tr><tr><td>Trichloro(1H,1H,2H,2H-perfluorooctyl)silane</td><td>Sigma-Aldritch</td><td>448931</td><td>Reacts violently with water</td></tr><tr><td>Ethanol, 200 proof Decon Labs</td><td>Fisher Scientific</td><td>04-355-223</td><td></td></tr><tr><td>Acetone</td><td>Fisher Scientific</td><td>A18P-4</td><td></td></tr><tr><td>Bel-Art 42025 Plastic Dessicator</td><td>Cole-Parmer</td><td>EW-06514-30</td><td></td></tr><tr><td>Epilog Fusion Laser Cutter, 120 W</td><td>Epilog Laser</td><td>Epilog Fusion M2 32 Laser</td><td></td></tr><tr><td>Isotemp Stirring Hotplate</td><td>Fisher Scientific</td><td>SP88850200</td><td></td></tr><tr><td>Ateco 14111 1.5 inch stainless steel cutter</td><td>Ateco</td><td>14111</td><td></td></tr><tr><td>Pyrex Glass Cell Culture Dish</td><td>Fisher Scientific</td><td>08-747B</td><td></td></tr><tr><td>Radio Frequency Plasma Cleaner</td><td>Harrick Plasma</td><td>PDC-32G</td><td>Used with Oxygen gas</td></tr><tr><td>Black Hole Laboratories Digivac</td><td>Black Hole Laboratories</td><td>Model 215</td><td></td></tr><tr><td>Intelli-Ray Ultraviolet Oven</td><td>Uvitron</td><td>UVO338</td><td></td></tr><tr><td>Compact Spin Coater</td><td>MTI Corporation</td><td>VTC-100A</td><td></td></tr><tr><td>Fisher Brand Isotemp Oven</td><td>Fisher Scientific</td><td>15-103-0510</td><td>Forced Air Convection</td></tr><tr><td>Gilson Positive Displacement Pipette P1000</td><td>Fisher Scientific</td><td>FD10006G</td><td></td></tr><tr><td>HeraCell VIOS 160i</td><td>Fisher Scientific</td><td>13 998 212PM</td><td></td></tr></tbody></table>

fabrication of refractive-index-matched devices for biomedical microfluidics

微流控装置的使用已成为生物医学应用的一个定义工具。当结合现代显微技术, 这些设备可以实现作为一个强大的平台的一部分, 能够作出同时互补测量。这两种技术的结合所产生的主要挑战是传统上用于制造微流控器件的材料与通常用于生物医学的水溶液的折射率不匹配。这种不匹配会在通道或设备边缘附近创建光学工件。一种解决方案是通过使用含氟聚合物 (如 MY133-V2000 的折射率与水 (n = 1.33) 类似) 来减少用于制造装置的材料的折射率。在这里, MY133-V2000 用软光刻技术制造出的微流控装置, 利用 O2等离子与丙烯酸保持架结合, 提高了 MY133-V2000 制造装置与烷基板。然后用细胞培养培养基填充24小时来测试该装置, 以证明该装置在典型的成像实验过程中保持细胞培养条件的能力。最后, 定量相镜 (QPM) 用于测量微通道内活黏附细胞内的质量分布。这样, 就能通过从低折射率聚合物 (如 MY133-V2000) 中制备器件来代替传统的软光刻材料 (如。总的来说, 这种制造微流控器件的方法可以很容易地集成到现有的软光刻工作流中, 以减少光学工件, 提高测量精度。

本协议描述了 MY133-V2000 的制备, 这种聚合物具有低折射率与水相匹配。该协议的一个关键特点是如何克服在使用氧等离子体的情况下缺乏粘附性, 并通过在丙烯酸支架内制造装置来提供密封通道所需的额外机械力。对基板 (图 1)。最终器件的低折射率在宏观尺度上明显显示 (图 2)。由这种材料制成的通道的边缘在空气中明显可见 ...

Watch this Scientific Journal Video about Fabrication of Refractive-index-matched Devices for Biomedical Microfluidics at JoVE.com

Fabrication of Refractive-index-matched Devices for Biomedical Microfluidics

该协议描述了微流控器件从 MY133-V2000 制造, 以消除由于微通道结构与水溶液之间的折射率不匹配而经常出现在微中的工件。该协议使用丙烯酸的持有人压缩封装的设备, 提高附着力的化学和机械。

生物医学微流体折射率匹配器件的研制

The use of microfluidic devices has emerged as a defining tool for biomedical applications. When combined with modern microscopy techniques, these devices ...

fabrication-refractive-index-matched-devices-for-biomedical

Universidad San Sebastian

Research

JoVE Journal

Engineering

7.4K 次观看.  University of Utah. 该协议描述了微流控器件从 MY133-V2000 制造, 以消除由于微通道结构与水溶液之间的折射率不匹配而经常出现在微中的工件。该协议使用丙烯酸的持有人压缩封装的设备, 提高附着力的化学和机械。

视频: Fabrication of Refractive-index-matched Devices for Biomedical Microfluidics

Fabrication of Silica Ultra High Quality Factor Microresonators

Whispering gallery resonant cavities confine light in circular orbits at their periphery.1-2 The photon storage lifetime in the cavity, quantified by the quality factor (Q) of the cavity, can be in excess of 500ns for cavities with Q factors above 100 million. As a result of their low material losses, silica microcavities have demonstrated some of the longest photon lifetimes to date1-2. Since a portion of the circulating light extends outside the resonator, these devices can also be used to probe the surroundings. This interaction has enabled numerous experiments in biology, such as single molecule biodetection and antibody-antigen kinetics, as well as discoveries in other fields, such as development of ultra-low-threshold microlasers, characterization of thin films, and cavity quantum electrodynamics studies.3-7
The two primary silica resonant cavity geometries are the microsphere and the microtoroid. Both devices rely on a carbon dioxide laser reflow step to achieve their ultra-high-Q factors (Q&gt;100 million).1-2,8-9 However, there are several notable differences between the two structures. Silica microspheres are free-standing, supported by a single optical fiber, whereas silica microtoroids can be fabricated on a silicon wafer in large arrays using a combination of lithography and etching steps. These differences influence which device is optimal for a given experiment.
Here, we present detailed fabrication protocols for both types of resonant cavities. While the fabrication of microsphere resonant cavities is fairly straightforward, the fabrication of microtoroid resonant cavities requires additional specialized equipment and facilities (cleanroom). Therefore, this additional requirement may also influence which device is selected for a given experiment.
Introduction
An optical resonator efficiently confines light at specific wavelengths, known as the resonant wavelengths of the device. 1-2 The common figure of merit for these optical resonators is the quality factor or Q. This term describes the photon lifetime (&tau;o) within the resonator, which is directly related to the resonator's optical losses. Therefore, an optical resonator with a high Q factor has low optical losses, long photon lifetimes, and very low photon decay rates (1/&tau;o). As a result of the long photon lifetimes, it is possible to build-up extremely large circulating optical field intensities in these devices. This very unique property has allowed these devices to be used as laser sources and integrated biosensors.10
A unique sub-class of resonators is the whispering gallery mode optical microcavity. In these devices, the light is confined in circular orbits at the periphery. Therefore, the field is not completely confined within the device, but evanesces into the environment. Whispering gallery mode optical cavities have demonstrated some of the highest quality factors of any optical resonant cavity to date.9,11 Therefore, these devices are used throughout science and engineering, including in fundamental physics studies and in telecommunications as well as in biodetection experiments. 3-7,12
Optical microcavities can be fabricated from a wide range of materials and in a wide variety of geometries. A few examples include silica and silicon microtoroids, silicon, silicon nitride, and silica microdisks, micropillars, and silica and polymer microrings.13-17 The range in quality factor (Q) varies as dramatically as the geometry. Although both geometry and high Q are important considerations in any field, in many applications, there is far greater leverage in boosting device performance through Q enhancement. Among the numerous options detailed previously, the silica microsphere and the silica microtoroid resonator have achieved some of the highest Q factors to date.1,9 Additionally, as a result of the extremely low optical loss of silica from the visible through the near-IR, both microspheres and microtoroids are able to maintain their Q factors over a wide range of testing wavelengths.18 Finally, because silica is inherently biocompatible, it is routinely used in biodetection experiments.
In addition to high material absorption, there are several other potential loss mechanisms, including surface roughness, radiation loss, and contamination loss.2 Through an optimization of the device size, it is possible to eliminate radiation losses, which arise from poor optical field confinement within the device. Similarly, by storing a device in an appropriately clean environment, contamination of the surface can be minimized. Therefore, in addition to material loss, surface scattering is the primary loss mechanism of concern.2,8
In silica devices, surface scattering is minimized by using a laser reflow technique, which melts the silica through surface tension induced reflow. While spherical optical resonators have been studied for many years, it is only with recent advances in fabrication technologies that researchers been able to fabricate high quality silica optical toroidal microresonators (Q&gt;100 million) on a silicon substrate, thus paving the way for integration with microfluidics.1
The present series of protocols details how to fabricate both silica microsphere and microtoroid resonant cavities. While silica microsphere resonant cavities are well-established, microtoroid resonant cavities were only recently invented.1 As many of the fundamental methods used to fabricate the microsphere are also used in the more complex microtoroid fabrication procedure, by including both in a single protocol it will enable researchers to more easily trouble-shoot their experiments.

We describe the use of a carbon dioxide laser reflow technique to fabricate silica resonant cavities, including free-standing microspheres and on-chip microtoroids. The reflow method removes surface imperfections, allowing long photon lifetimes within both devices. The resulting devices have ultra high quality factors, enabling applications ranging from telecommunications to biodetection.

硅胶超高品质因子微谐振器的制造

Whispering gallery resonant cavities confine light in circular orbits at their periphery.1-2 The photon storage lifetime in the cavity, quantified by the ...

科研

工程学

Fabrication and Testing of Microfluidic Optomechanical Oscillators

Cavity optomechanics experiments that parametrically couple the phonon modes and photon modes have been investigated in various optical systems including microresonators. However, because of the increased acoustic radiative losses during direct liquid immersion of optomechanical devices, almost all published optomechanical experiments have been performed in solid phase. This paper discusses a recently introduced hollow microfluidic optomechanical resonator. Detailed methodology is provided to fabricate these ultra-high-Q microfluidic resonators, perform optomechanical testing, and measure radiation pressure-driven breathing mode and SBS-driven whispering gallery mode parametric vibrations. By confining liquids inside the capillary resonator, high mechanical- and optical- quality factors are simultaneously maintained.

Parametric optomechanical excitations have recently been experimentally demonstrated in microfluidic optomechanical resonators by means of optical radiation pressure and stimulated Brillouin scattering. This paper describes the fabrication of these microfluidic resonators along with methodologies for generating and verifying optomechanical oscillations.

制造和微流控光机振荡器测试

Cavity optomechanics experiments that parametrically couple the phonon modes and photon modes have been investigated in various optical systems including ...

Endothelialized Microfluidics for Studying Microvascular Interactions in Hematologic Diseases

Advances in microfabrication techniques have enabled the production of inexpensive and reproducible microfluidic systems for conducting biological and biochemical experiments at the micro- and nanoscales 1,2. In addition, microfluidics have also been specifically used to quantitatively analyze hematologic and microvascular processes, because of their ability to easily control the dynamic fluidic environment and biological conditions3-6. As such, researchers have more recently used microfluidic systems to study blood cell deformability, blood cell aggregation, microvascular blood flow, and blood cell-endothelial cell interactions6-13.However, these microfluidic systems either did not include cultured endothelial cells or were larger than the sizescale relevant to microvascular pathologic processes. A microfluidic platform with cultured endothelial cells that accurately recapitulates the cellular, physical, and hemodynamic environment of the microcirculation is needed to further our understanding of the underlying biophysical pathophysiology of hematologic diseases that involve the microvasculature.
Here, we report a method to create an "endothelialized" in vitro model of the microvasculature, using a simple, single mask microfabrication process in conjunction with standard endothelial cell culture techniques, to study pathologic biophysical microvascular interactions that occur in hematologic disease. This "microvasculature-on-a-chip" provides the researcher with a robust assay that tightly controls biological as well as biophysical conditions and is operated using a standard syringe pump and brightfield/fluorescence microscopy. Parameters such as microcirculatory hemodynamic conditions, endothelial cell type, blood cell type(s) and concentration(s), drug/inhibitory concentration etc., can all be easily controlled. As such, our microsystem provides a method to quantitatively investigate disease processes in which microvascular flow is impaired due to alterations in cell adhesion, aggregation, and deformability, a capability unavailable with existing assays.

A method to culture an endothelial cell monolayer throughout the entire inner 3D surface of a microfluidic device with microvascular-sized channels (&lt;30 &mu;m) is described. This in vitro microvasculature model enables the study of biophysical interactions between blood cells, endothelial cells, and soluble factors in hematologic diseases.

内皮化微流体研究在血液病的微血管相互作用

Advances in  microfabrication techniques have enabled the production of inexpensive and  reproducible microfluidic systems for conducting biological and ...

生物工程

Fabrication and Operation of Acoustofluidic Devices Supporting Bulk Acoustic Standing Waves for Sheathless Focusing of Particles

Acoustophoresis refers to the displacement of suspended objects in response to directional forces from sound energy. Given that the suspended objects must be smaller than the incident wavelength of sound and the width of the fluidic channels are typically tens to hundreds of micrometers across, acoustofluidic devices typically use ultrasonic waves generated from a piezoelectric transducer pulsating at high frequencies (in the megahertz range). At characteristic frequencies that depend on the geometry of the device, it is possible to induce the formation of standing waves that can focus particles along desired fluidic streamlines within a bulk flow. Here, we describe a method for the fabrication of acoustophoretic devices from common materials and clean room equipment. We show representative results for the focusing of particles with positive or negative acoustic contrast factors, which move towards the pressure nodes or antinodes of the standing waves, respectively. These devices offer enormous practical utility for precisely positioning large numbers of microscopic entities (e.g., cells) in stationary or flowing fluids for applications ranging from cytometry to assembly.

Acoustofluidic devices use ultrasonic waves within microfluidic channels to manipulate, concentrate and isolate suspended micro and nanoscopic entities. This protocol describes the fabrication and operation of such a device supporting bulk acoustic standing waves to focus particles in a central streamline without the aid of sheath fluids.

制造和Acoustofluidic设备的运营支撑体声波驻波的聚焦鞘颗粒

Acoustophoresis refers to the displacement of suspended objects in response to directional forces from sound energy. Given that the suspended objects must ...

One-Step Approach to Fabricating Polydimethylsiloxane Microfluidic Channels of Different Geometric Sections by Sequential Wet Etching Processes

Polydimethylsiloxane (PDMS) materials are substantially exploited to fabricate microfluidic devices by using soft lithography replica molding techniques. Customized channel layout designs are necessary for specific functions and integrated performance of microfluidic devices in numerous biomedical and chemical applications (e.g., cell culture, biosensing, chemical synthesis, and liquid handling). Owing to the nature of molding approaches using silicon wafers with photoresist layers patterned by photolithography as master molds, the microfluidic channels commonly have regular cross sections of rectangular shapes with identical heights. Typically, channels with multiple heights or different geometric sections are designed to possess particular functions and to perform in various microfluidic applications (e.g., hydrophoresis is used for sorting particles and in continuous flows for separating blood cells6,7,8,9). Therefore, a great deal of effort has been made in constructing channels with various sections through multiple-step approaches like photolithography using several photoresist layers and assembly of different PDMS thin sheets. Nevertheless, such multiple-step approaches usually involve tedious procedures and extensive instrumentation. Furthermore, the fabricated devices may not perform consistently and the resulted experimental data may be unpredictable. Here, a one-step approach is developed for the straightforward fabrication of microfluidic channels with different geometric cross sections through PDMS sequential wet etching processes, that introduces etchant into channels of planned single-layer layouts embedded in PDMS materials. Compared to the existing methods for manufacturing PDMS microfluidic channels with different geometries, the developed one-step approach can significantly simplify the process to fabricate channels with non-rectangular sections or various heights. Consequently, the technique is a way of constructing complex microfluidic channels, which provides a fabrication solution for the advancement of innovative microfluidic systems.

Several methods are available for the fabrication of channels of non-rectangular sections embedded in polydimethylsiloxane microfluidic devices. Most of them involve multistep manufacturing and extensive alignment. In this paper, a one-step approach is reported for fabricating microfluidic channels of different geometric cross sections by polydimethylsiloxane sequential wet etching.

连续湿法刻蚀法制作不同几何断面烷微流控通道的一步法

Polydimethylsiloxane (PDMS) materials are substantially exploited to fabricate microfluidic devices by using soft lithography replica molding techniques. ...

Design and Development of a Three-Dimensionally Printed Microscope Mask Alignment Adapter for the Fabrication of Multilayer Microfluidic Devices

This project aims to develop an easy-to-use and cost-effective platform for the fabrication of precise, multilayer microfluidic devices, which typically can only be achieved using costly equipment in a clean room setting. The key part of the platform is a three dimensionally (3D) printed microscope mask alignment adapter (MMAA) compatible with regular optical microscopes and ultraviolet (UV) light exposure systems. The overall process of creating the device has been vastly simplified because of the work done to optimize the device design. The process entails finding the proper dimensions for the equipment available in the laboratory and 3D-printing the MMAA with the optimized specifications. Experimental results show that the optimized MMAA designed and manufactured by 3D printing performs well with a common microscope and light exposure system. Using a master mold prepared by the 3D-printed MMAA, the resulting microfluidic devices with multilayered structures contain alignment errors of &lt;10 &#181;m, which is sufficient for common microchips. Although human error through transportation of the device to the UV light exposure system can cause larger fabrication errors, the minimal errors achieved in this study are attainable with practice and care. Furthermore, the MMAA can be customized to fit any microscope and UV exposure system by making changes to the modeling file in the 3D printing system. This project provides smaller laboratories with a useful research tool as it only requires the use of equipment that is typically already available to laboratories that produce and use microfluidic devices. The following detailed protocol outlines the design and 3D printing process for the MMAA. In addition, the steps for procuring a multilayer master mold using the MMAA and producing poly(dimethylsiloxane) (PDMS) microfluidic chips is also described herein.

This project allows small laboratories to develop an easy-to-use platform for the fabrication of precise multilayer microfluidic devices. The platform consists of a three-dimensionally printed microscope mask alignment adapter using which multilayer microfluidic devices with alignment errors of &lt;10 &#181;m were achieved.

设计和开发用于制造多层微流体设备的三维打印显微镜面膜对齐适配器

This project aims to develop an easy-to-use and cost-effective platform for the fabrication of precise, multilayer microfluidic devices, which typically ...

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 ...

Computer Numerical Control Micromilling of a Microfluidic Acrylic Device with a Staggered Restriction for Magnetic Nanoparticle-Based Immunoassays

Microfluidic systems have greatly improved immunoassay techniques. However, many microfabrication techniques require specialized, expensive, or complicated equipment, making fabrication costly and incompatible with mass production, which is one of the most important preconditions for point-of-care tests (POCT) to be adopted in low-resource settings. This work describes the fabrication process of an acrylic (polymethylmethacrylate, PMMA) device for nanoparticle-conjugated enzymatic immunoassay testing using the computer numerical control (CNC) micromilling technique. The functioning of the microfluidic device is shown by performing an immunoassay to detect a commercial antibody using lysozyme as a model antigen conjugated to 100 nm magnetic nanoparticles. This device integrates a physical staggered restriction of only 5 &#181;m in height, used to capture magnetic microparticles that make up a magnetic trap by placing an external magnet. In this way, the magnetic force on the immunosupport of conjugated nanoparticles is enough to capture them and resist flow drag. This microfluidic device is particularly suitable for low-cost mass production without the loss of precision for immunoassay performance.

Microfluidics is a powerful tool for the development of diagnostic tests. However, expensive equipment and materials, as well as laborious fabrication and handling techniques, are often required. Here, we detail the fabrication protocol of an acrylic microfluidic device for magnetic micro- and nanoparticle-based immunoassays in a low-cost and simple-to-use setting.

基于磁性纳米颗粒免疫分析的交错限制微流控亚克力装置的计算机数控微铣削

Microfluidic systems have greatly improved immunoassay techniques. However, many microfabrication techniques require specialized, expensive, or ...

Soft Lithographic Procedure for Producing Plastic Microfluidic Devices with View-ports Transparent to Visible and Infrared Light

Infrared (IR) spectro-microscopy of living biological samples is hampered by the absorption of water in the mid-IR range and by the lack of suitable microfluidic devices. Here, a protocol for the fabrication of plastic microfluidic devices is demonstrated, where soft lithographic techniques are used to embed transparent Calcium Fluoride (CaF2) view-ports in connection with observation chamber(s). The method is based on a replica casting approach, where a polydimethylsiloxane (PDMS) mold is produced through standard lithographic procedures and then used as the template to produce a plastic device. The plastic device features ultraviolet/visible/infrared (UV/Vis/IR) -transparent windows made of CaF2 to allow for direct observation with visible and IR light. The advantages of the proposed method include: a reduced need for accessing a clean room micro-fabrication facility, multiple view-ports, an easy and versatile connection to an external pumping system through the plastic body, flexibility of the design, e.g., open/closed channels configuration, and the possibility to add sophisticated features such as nanoporous membranes.

A protocol for the fabrication of plastic microfluidic devices with transparent view-ports for visible and infrared light imaging is described.

生产塑料微流控装置与查看端口对可见光和红外透明软光刻过程

Infrared (IR) spectro-microscopy of living biological samples is hampered by the absorption of water in the mid-IR range and by the lack of suitable ...

Rapid Fabrication of Custom Microfluidic Devices for Research and Educational Applications

Microfluidic devices allow for the manipulation of fluids, particles, cells, micro-sized organs or organisms in channels ranging from the nano to submillimeter scales. A rapid increase in the use of this technology in the biological sciences has prompted a need for methods that are accessible to a wide range of research groups. Current fabrication standards, such as PDMS bonding, require expensive and time consuming lithographic and bonding techniques. A viable alternative is the use of equipment and materials that are easily affordable, require minimal expertise and allow for the rapid iteration of designs. In this work we describe a protocol for designing and producing PET-laminates (PETLs), microfluidic devices that are inexpensive, easy to fabricate, and consume significantly less time to generate than other approaches to microfluidics technology. They consist of thermally bonded film sheets, in which channels and other features are defined using a craft cutter. PETLs solve field-specific technical challenges while dramatically reducing obstacles to adoption. This approach facilitates the accessibility of microfluidics devices in both research and educational settings, providing a reliable platform for new methods of inquiry.

Here we present a protocol to design and fabricate custom microfluidic devices with minimal financial and time investment. The aim is to facilitate the adoption of microfluidic technologies in biomedical research laboratories and educational settings.