To characterize responses to different angles of polarization, we rotated the polarizer discontinuously in 30° steps with the UV LED on throughout. Each of the 12 positions (six unique angles of polarization) was maintained for 4–4.5 s and we used 4 s of imaging data collected during this period in our analysis. With a volume-rate of at least 1 Hz, this ensured that at least four time-points were captured for every plane in the imaging volume. The polarizer was then rotated through 30° in 0.5 s. At least two complete revolutions of the polarizer were made. For recordings with the polarizer removed, the procedure was repeated and one revolution of the stimulus was made. The discontinuous rotation of the polarizer differs from most previous studies of polarization-sensitive neurons in insects, which have typically analyzed spike frequency modulated by continuously rotating stimuli, and as such the data obtained using this protocol may not be directly comparable.

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