Paper-Supported High-Throughput 3D Culturing, Trapping, and Monitoring of Caenorhabditis Elegans
Paper-Supported High-Throughput 3D Culturing, Trapping, and Monitoring of Caenorhabditis Elegans
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DOI：
10.3390/mi11010099
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发表时间：
2020-01-01
期刊：
影响因子：
3.4
通讯作者：
Choi, Seokheun
中科院分区：
文献类型：
--
作者：
Tahernia, Mehdi;Mohammadifar, Maedeh;Choi, Seokheun
我们开发了一种创新的纸基平台，用于高通量培养、捕获和监测C。优雅一个96孔阵列很容易通过将营养补充的纸基板上的微加工96孔塑料框架，提供高通量的三维培养环境和原位分析的蠕虫。本文允许C.线虫通过多孔和水生纸基质，直到蠕虫生长并达到下一个发育阶段，其增加的身体大小与纸孔相当。当C.当线虫变得大于纸基质的孔径时，蠕虫被捕获并固定用于进一步的高通量成像和分析。这项工作将提供一个简单而强大的技术，高通量排序和监测的C。通过控制和选择不同孔径的纸张，此外，我们开发了另一种类型的3D培养系统，通过使用纸状透明聚碳酸酯基板进行更高分辨率的成像。该设备使用聚碳酸酯层的多层结构作为支架，以模拟蠕虫的3D自然栖息地。由于基板薄，机械强度高，而且大部分是多孔的，所以层状结构允许C。elegans的三维运动和行为自由，促进了C. elegans及其主要食物E.杆菌这种结构的透明度有助于在显微镜下观察蠕虫。发育、育性和动态行为。3D培养平台中的秀丽隐杆线虫的表现优于标准2D培养技术。
We developed an innovative paper-based platform for high-throughput culturing, trapping, and monitoring of C. elegans. A 96-well array was readily fabricated by placing a nutrient-replenished paper substrate on a micromachined 96-well plastic frame, providing high-throughput 3D culturing environments and in situ analysis of the worms. The paper allows C. elegans to pass through the porous and aquatic paper matrix until the worms grow and reach the next developmental stages with the increased body size comparable to the paper pores. When the diameter of C. elegans becomes larger than the pore size of the paper substrate, the worms are trapped and immobilized for further high-throughput imaging and analysis. This work will offer a simple yet powerful technique for high-throughput sorting and monitoring of C. elegans at a different larval stage by controlling and choosing different pore sizes of paper. Furthermore, we developed another type of 3D culturing system by using paper-like transparent polycarbonate substrates for higher resolution imaging. The device used the multi-laminate structure of the polycarbonate layers as a scaffold to mimic the worm's 3D natural habitats. Since the substrate is thin, mechanically strong, and largely porous, the layered structure allowed C. elegans to move and behave freely in 3D and promoted the efficient growth of both C. elegans and their primary food, E. coli. The transparency of the structure facilitated visualization of the worms under a microscope. Development, fertility, and dynamic behavior of C. elegans in the 3D culture platform outperformed those of the standard 2D cultivation technique.