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Experiments on the flow-sensitive lateral line system of fishes have provided important insights into the function and sensory transduction of vertebrate hair cells. A common experimental approach has been to pharmacologically block lateral line hair cells and measure how behavior changes. Cobalt chloride (CoCl2) blocks the lateral line by inhibiting calcium movement through the membrane channels of hair cells, but high concentrations can be toxic, making it unclear whether changes in behavior are due to a blocked lateral line or poor health. Here, we identify a non-toxic treatment of cobalt that completely blocks lateral line hair cells. We exposed 5-day post fertilization zebrafish larvae to CoCl2 concentrations ranging from 1 to 20 mM for 15 min and measured 1) the spiking rate of the afferent neurons contacting hair cells and 2) the larvae's health and long-term survival. Our results show that a 15-min exposure to 5 mM CoCl2 abolishes both spontaneous and evoked afferent firing. This treatment does not change swimming behavior, and results in >85% survival after 5 days. Weaker treatments of CoCl2 did not eliminate afferent activity, while stronger treatments caused close to 50% mortality. Our work provides a guideline for future zebrafish investigations where physiological confirmation of a blocked lateral line system is required.The effect of cobalt chloride on the sensitivity of lateral line hair cells and the health of fish was explored.A 15-min exposure to 5 mM cobalt chloride is non-toxic and completely blocks hair cells.Lateral line afferent neurons still respond to stimulation after cobalt chloride treatments below 5 mM.Treatments of cobalt chloride above 5 mM cause significant mortality.These results will guide future work requiring a safe and verified treatment for blocking hair cells in zebrafish.