Max Planck Institute for Biophysical Chemistry, Am Fassberg 11, 37077 Göttingen, Germany.
J Neurosci Methods. 2011 Sep 30;201(1):78-88. doi: 10.1016/j.jneumeth.2011.07.013. Epub 2011 Jul 23.
Caenorhabditis elegans larvae display specific behavior and development that is not observed in adults. For example, larvae go through a molting cycle that includes a sleep-like state prior to the molt. The study of these processes requires high-resolution long-term observation of individual animals. Here we describe a method for simultaneous culture and observation of several individual young C. elegans larvae inside agarose hydrogel-based arrayed microcompartments. We used agarose hydrogel microcompartments to observe and quantify larval specific sleep-wake-like behavior and to observe neuronal rewiring using confocal fluorescence microscopy without acute immobilization. We found no behavioral aberrations caused by area restriction. We show that worms cultured inside hydrogel microcompartments develop into normal adults. Thus, hydrogel microcompartments appear useful for long-term observation of larval behavior and development.
秀丽隐杆线虫幼虫表现出特定的行为和发育，而这些在成虫中是观察不到的。例如，幼虫经历一个蜕皮周期，在蜕皮前有一个类似于睡眠的状态。这些过程的研究需要对单个动物进行高分辨率的长期观察。在这里，我们描述了一种在琼脂糖水凝胶基阵列微隔间内同时培养和观察几个个体年轻秀丽隐杆线虫幼虫的方法。我们使用琼脂糖水凝胶微隔间来观察和量化幼虫特定的睡眠-觉醒样行为，并使用共聚焦荧光显微镜进行神经元重布线观察，而无需急性固定。我们发现区域限制不会引起行为异常。我们表明，在水凝胶微隔间中培养的蠕虫发育成正常的成虫。因此，水凝胶微隔间似乎可用于幼虫行为和发育的长期观察。
