Microfluidic Transportation Control of Larval Zebrafish through Optomotor Regulations under a Pressure-Driven Flow

To perform zebrafish larvae-related experiments within a microfluidic environment, the larvae need to be anesthetized and subsequently transported into respective test sections through mechanical or manual means. However, anesthetization tends to affect larval sensory perceptions, hindering their natural behaviors. Taking into account that juvenile larvae move naturally within their environment by accessing visual as well as hydromechanical cues, this work proposes an experimental framework to transport nonanesthetized larvae within a microfluidic environment by harmonically tuning both of the aforementioned cues. To provide visual cues, computer-animated moving gratings were provided through an in-house-developed control interface that drove the larval optomotor response. In the meantime, to provide hydromechanical cues, the flow rate was tuned using a syringe pump that affected the zebrafish larvae's lateral line movement. The results obtained (corresponding to different test conditions) suggest that the magnitude of both modalities plays a crucial role in larval transportation and orientation control. For instance, with a flow rate tuning of 0.1 mL/min along with grating parameters of 1 Hz temporal frequency, the average transportation time for larvae that were 5 days postfertilization was recorded at 1.29 ± 0.49 s, which was approximately three times faster than the transportation time required only in the presence of hydromechanical cues.