Ghaemi Reza, Rezai Pouya, Iyengar Balaji G, Selvaganapathy Ponnambalam Ravi
Department of Mechanical Engineering, McMaster University, Hamilton, ON, Canada.
Lab Chip. 2015 Feb 21;15(4):1116-22. doi: 10.1039/c4lc01245c.
Two microfluidic devices (pneumatic chip and FlexiChip) have been developed for immobilization and live-intact fluorescence functional imaging of Drosophila larva's Central Nervous System (CNS) in response to controlled acoustic stimulation. The pneumatic chip is suited for automated loading/unloading and potentially allows high throughput operation for studies with a large number of larvae while the FlexiChip provides a simple and quick manual option for animal loading and is suited for smaller studies. Both chips were capable of significantly reducing the endogenous CNS movement while still allowing the study of sound-stimulated CNS activities of Drosophila 3rd instar larvae using genetically encoded calcium indicator GCaMP5. Temporal effects of sound frequency (50-5000 Hz) and intensity (95-115 dB) on CNS activities were investigated and a peak neuronal response of 200 Hz was identified. Our lab-on-chip devices can not only aid further studies of Drosophila larva's auditory responses but can be also adopted for functional imaging of CNS activities in response to other sensory cues. Auditory stimuli and the corresponding response of the CNS can potentially be used as a tool to study the effect of chemicals on the neurophysiology of this model organism.
已开发出两种微流控装置（气动芯片和柔性芯片），用于对果蝇幼虫中枢神经系统（CNS）进行固定和完整活体荧光功能成像，以响应受控的声学刺激。气动芯片适用于自动加载/卸载，可能允许对大量幼虫进行高通量操作，而柔性芯片为动物加载提供了一种简单快捷的手动选择，适用于较小规模的研究。两种芯片都能够显著减少内源性中枢神经系统的运动，同时仍能使用基因编码的钙指示剂GCaMP5研究果蝇三龄幼虫受声音刺激的中枢神经系统活动。研究了声音频率（50 - 5000 Hz）和强度（95 - 115 dB）对中枢神经系统活动的时间效应，并确定了200 Hz的峰值神经元反应。我们的芯片实验室装置不仅有助于进一步研究果蝇幼虫的听觉反应，还可用于响应其他感官线索的中枢神经系统活动的功能成像。听觉刺激和中枢神经系统的相应反应有可能用作研究化学物质对这种模式生物神经生理学影响的工具。
