Animals often select for habitats that increase their chance of survival by balancing the need to acquire food, reproduce and avoid predation. Perennial blooms of golden jellyfish (Mastigias papua etpisoni) are present in Jellyfish Lake, Palau, a popular tourist destination. Based on the species’ economic importance and unusual behavioural complexity, increased understanding of jellyfish habitat selection is necessary. We used a novel approach, a REMUS autonomous underwater vehicle, to quantify jellyfish distribution, abundance and habitat, and compared these findings to traditional methods. Midday acoustic surveys showed jellyfish distribution was patchy and the population resided mainly on the eastern side of the lake, as it is known that jellyfish migrate eastward towards the sun. Highest vertical densities of jellyfish were at 6–7 m, potentially to mitigate UV damage or photoinhibition of their photosymbionts, suggesting a coupling exists between their vertical distribution and water properties. Abundance estimates of jellyfish were ~2.75 and ~7.1 million (~2 million excluding bell diameters <1 cm) from acoustic and net samples, suggesting the methodology employed underestimated the population's smaller size fraction and non-synoptic surveys could impact estimates due to unresolved patchiness. Our approach could investigate population dynamics, behaviour or habitat associations on fine scales.