Journal: iScience
Article Title: JACUZI-SD: An automated, high-throughput, minimally stressful approach to sleep depriving larval zebrafish
doi: 10.1016/j.isci.2026.115954
Figure Lengend Snippet: JACUZI-SD diagram (A) Arduino UNO wired to Raspberry Pi. Pulse duration and inter-pulse interval are randomized to minimize habituation to the stimulus. Pulse duration increases and inter-pulse interval decreases with each hour of sleep deprivation to combat increasing sleep pressure and minimize habituation. (B) Submersible water pump in a beaker of E3 attached to (C) nontoxic plastic tubing, which is threaded through the back panel of the ZebraBox, and the gap generated is tightly covered with light-blocking fabric. (D) Within the ZebraBox, infrared illumination allows high-resolution camera to record sleep behavior of 96 larvae individually housed in a mesh-bottom 96-well plate secured into the milli-fluidic device base. The water bath sits on top of an IR and LED light source to provide 14 h:10 h light:dark cycles. (E) 3D-printed mesh bottom 96-well plate adapted from Kroll et al., 2025. (F) Stereolithography-printed milli-fluidic device secured beneath the 96-well plate provides channels to deliver water pulses beneath each larva. The entire arena is symmetrical to prevent any baseline behavioral differences across the plate. Stainless steel adapters screw into the milli-fluidic device and the tubing is attached to the barbs of these adapters. Parafilm is used around this junction to minimize loss of water flow. (G) Epoxy border holds both plates together and prevents water disturbances on control side of plate.
Article Snippet: The network was printed using a commercial stereolithography 3D printer (Form 3, Formlabs, USA) using standard resin (Clear v4, Formlabs, USA) with a 100μm layer height.
Techniques: Generated, Blocking Assay, Control