Department of Biomedical Engineering, Washington University in St. Louis, USA.
Lab Chip. 2017 Nov 7;17(22):3909-3920. doi: 10.1039/c7lc00916j.
We have miniaturized standard culture techniques to rear arrays of isolated, individual C. elegans throughout their lives on solid gel media. The resulting apparatus is compatible with brightfield and fluorescence microscopy, enabling longitudinal studies of morphology and fluorescent transgene expression. Our culture system exploits a novel crosslinking reaction between a polyethylene glycol hydrogel and a silicone elastomer to constrain animals to individual "corrals" on the gel surface. These devices are simple to construct on the benchtop with commercially available reagents, and, unlike microfluidic isolation methods, do not rely on micropatterned materials. We demonstrate that this new culture method has negligible effects on the physiology of C. elegans compared to standard culture on agar plates. In addition, RNAi techniques are effective in this system. Finally, the hydrogel-silicone binding chemistry that we developed also allows traditional microfluidic devices to be covalently attached to gel substrates instead of glass.
我们已经将标准的培养技术小型化，以便在固体凝胶培养基上终生饲养成群的、单独的秀丽隐杆线虫。由此产生的设备与明场和荧光显微镜兼容，可进行形态和荧光转基因表达的纵向研究。我们的培养系统利用聚乙二醇水凝胶和硅橡胶之间的新型交联反应将动物限制在凝胶表面的单个“畜栏”中。这些设备可以使用市售试剂在台面上轻松构建，与微流控隔离方法不同，它们不依赖于微图案材料。与在琼脂平板上进行标准培养相比，我们证明这种新的培养方法对秀丽隐杆线虫的生理几乎没有影响。此外，RNAi 技术在该系统中也很有效。最后，我们开发的水凝胶-硅橡胶结合化学物质也允许将传统的微流控设备共价连接到凝胶基底上，而不是玻璃上。
