Fabrication of a microfluidic device for the compartmentalization of neuron soma and axons.
Fabrication of a microfluidic device for the compartmentalization of neuron soma and axons.
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DOI：
10.3791/261
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发表时间：
2007-01-01
期刊：
影响因子：
--
通讯作者：
Cotman, Carl
中科院分区：
文献类型：
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作者：
Harris, Joseph;Lee, Hyuna;Cotman, Carl
在这个视频中，我们展示了软光刻技术与聚二甲基硅氧烷（PDMS），我们使用它来制造一个微流体装置培养神经元。以前，使用SU-8和光刻法将硅晶片图案化为神经元微流体设备的设计，以创建主模具，或者我们简单地称为“母模”。接下来，我们将硅聚合物PDMS倒在母版的顶部，然后通过将PDMS加热到80摄氏度持续1小时来固化。PDMS形成装置的阴模。然后小心地切割PDMS并将其从母版上提起。在储液器所在位置打孔，并将多余的PDMS从器械上修剪掉。氮气用于吹走器械上的任何多余碎屑。此时，器械已准备就绪，可以使用等离子体灭菌器/清洁器将其粘合到康宁1号盖板玻璃上，也可以通过简单地将器械放置在盖板玻璃顶部将其可逆地粘合到盖板玻璃上。器械与玻璃的可逆粘合在单独的视频中介绍，首先需要使用70%乙醇或高压灭菌对器械进行灭菌。等离子体处理对器械进行灭菌，因此无需进一步处理。然而，当对器械进行等离子体处理时，重要的是在等离子体处理的10分钟内向器械中添加液体，同时表面仍然是亲水性的。等待超过10分钟才将液体添加到装置中会使液体难以进入装置。神经元装置通常与盖玻片等离子体结合，并立即将pH 8.5硼酸盐缓冲液中的0.5mg/ml聚-L-赖氨酸（PLL）加入到装置中。与PLL孵育至少3小时后，用dH 2 O水清洗器械至少3次，每次清洗之间至少间隔15分钟。接下来，去除水并将新鲜培养基添加到装置中。此时，器械已准备就绪。重要的是要记住，在这一点上，永远不要删除所有的媒体从设备。总是把媒体放在主频道。
In this video, we demonstrate the technique of soft lithography with polydimethyl siloxane (PDMS) which we use to fabricate a microfluidic device for culturing neurons. Previously, a silicon wafer was patterned with the design for the neuron microfluidic device using SU-8 and photolithography to create a master mold, or what we simply refer to as a "master". Next, we pour the silicon polymer PDMS on top of the master which is then cured by heating the PDMS to 80 degrees C for 1 hour. The PDMS forms a negative mold of the device. The PDMS is then carefully cut and lifted away from the master. Holes are punched where the reservoirs will be and the excess PDMS trimmed away from the device. Nitrogen is used to blow away any excess debris from the device. At this point the devices are now ready for use and can either bonded to corning No. 1 cover glass with a plasma sterilizer/cleaner or can be reversibly bound to the cover glass by simply placing the device on top of the cover glass. The reversible bonding of the device to glass is covered in a separate video and requires first that the device be sterilized either with 70% ethanol or by autoclaving. Plasma treating sterilizes the devices so no further treatment is necessary. It is, however, important, when plasma-treating the devices, to add liquid to the devices within 10 minutes of the plasma treatment while the surfaces are still hydrophilic. Waiting longer than 10 minutes to add liquid to the device makes it difficult for the liquid to enter the device. The neuron devices are typically plasma-bound to cover glass and 0.5 mg/ml poly-L-lysine (PLL) in pH 8.5 borate buffer is immediately added to the device. After a minimum of 3 hours incubating with PLL, the devices are washed with dH2O water a minimum of 3 times with at least 15 minutes between each wash. Next, the water is removed and fresh media is added to the device. At this point the device is ready for use. It is important to remember at this point to never remove all the media from the device. Always leave media in the main channel.