Laterally orienting C. elegans using geometry at microscale for high-throughput visual screens in neurodegeneration and neuronal development studies.
Laterally orienting C. elegans using geometry at microscale for high-throughput visual screens in neurodegeneration and neuronal development studies.
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DOI：
10.1371/journal.pone.0035037
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发表时间：
2012
期刊：
影响因子：
3.7
通讯作者：
Lu H
中科院分区：
文献类型：
--
作者：
Cáceres Ide C;Valmas N;Hilliard MA;Lu H
秀丽隐杆线虫由于其相对简单的神经系统、已测序的基因组以及大量转基因和突变菌株的可用性，是利用遗传学研究神经科学的优秀模型系统。近年来，微流控装置已被用于高通量遗传筛选，取代了手工处理秀丽隐杆线虫的传统方法。然而，微流控装置内线虫的方向是随机的，往往不利于检测，阻碍了视觉分析和整体吞吐量。此外，虽然以前的研究使用了偏向头部和尾部定向的方法，但现有的技术都不允许沿背-腹侧身体轴定向。在这里，我们设计了一种简单而稳健的方法，用于在微流体装置中被动地将蠕虫定向到侧体位置，以方便检查特定的背腹对齐形态特征。使用这种技术，我们可以将动物定位到横向方向，效率高达84%，而使用现有方法的效率为21%。我们分离了6个具有神经元发育或神经退行性缺陷的突变体，表明我们的技术可用于芯片上分析和高通量视觉屏幕。
C. elegans is an excellent model system for studying neuroscience using genetics because of its relatively simple nervous system, sequenced genome, and the availability of a large number of transgenic and mutant strains. Recently, microfluidic devices have been used for high-throughput genetic screens, replacing traditional methods of manually handling C. elegans. However, the orientation of nematodes within microfluidic devices is random and often not conducive to inspection, hindering visual analysis and overall throughput. In addition, while previous studies have utilized methods to bias head and tail orientation, none of the existing techniques allow for orientation along the dorso-ventral body axis. Here, we present the design of a simple and robust method for passively orienting worms into lateral body positions in microfluidic devices to facilitate inspection of morphological features with specific dorso-ventral alignments. Using this technique, we can position animals into lateral orientations with up to 84% efficiency, compared to 21% using existing methods. We isolated six mutants with neuronal development or neurodegenerative defects, showing that our technology can be used for on-chip analysis and high-throughput visual screens.
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