Caenorhabditis elegans is an important genetic model for neuroscience studies, used for analyses of how genes control connectivity, neuronal function, and behavior. To date, however, most studies of neuronal function in C. elegans are incapable of obtaining microscopy imaging with subcellular resolution and behavior analysis in the same set of animals. This constraint stems from the immobilization requirement for high-resolution imaging that is incompatible with behavioral analysis using conventional immobilization techniques. Here, we present a novel microfluidic device that uses surface acoustic waves (SAW) as a non-contact method to temporarily immobilize worms for a short period (30 seconds). We optimize the SAW based protocol for rapid switching between free-swimming and immobilized states, facilitating non-invasive analysis of swimming behavior as well as high-resolution synaptic imaging in the same animal. We find that the coupling of heat and acoustic pressure play a key role in the immobilization process. We introduce a proof-of-concept longitudinal study, illustrating that the device enables repeated imaging of fluorescently tagged synaptic receptors in command interneurons and analysis of swimming behavior in the same animals for three days. This longitudinal approach provides the first correlative analysis of synaptic glutamatergic receptors and swimming behavior in aging animals. We anticipate that this device will enable further longitudinal analysis of animal motility and subcellular morphological changes during development and aging in C. elegans.

中文翻译：

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用于重复和可逆临时固定线虫的表面声波微流体

秀丽隐杆线虫是神经科学研究的重要遗传模型，用于分析基因如何控制连通性、神经元功能和行为。然而，迄今为止，大多数关于秀丽隐杆线虫神经元功能的研究无法在同一组动物中获得具有亚细胞分辨率和行为分析的显微镜成像。这种限制源于高分辨率成像的固定要求，这与使用传统固定技术进行的行为分析不相容。在这里，我们提出了一种新型微流体装置，该装置使用表面声波 (SAW) 作为非接触方法在短时间内（30 秒）暂时固定蠕虫。我们优化了基于 SAW 的协议，以便在自由游泳和固定状态之间快速切换，促进游泳行为的非侵入性分析以及同一动物的高分辨率突触成像。我们发现热和声压的耦合在固定过程中起着关键作用。我们介绍了一项概念验证纵向研究，说明该设备能够对命令中间神经元中的荧光标记突触受体进行重复成像，并对同一动物的游泳行为进行为期三天的分析。这种纵向方法首次对衰老动物的突触谷氨酸能受体和游泳行为进行了相关分析。我们预计该设备将能够进一步纵向分析动物在发育和衰老过程中的运动性和亚细胞形态变化C. 线虫。